CN116568814A

CN116568814A - Supported antibodies and uses thereof

Info

Publication number: CN116568814A
Application number: CN202180074827.5A
Authority: CN
Inventors: C·巴恩斯; Y·夏尔马; S·多利夫; O·弗兰科尼
Original assignee: Cognate3 LLC
Current assignee: Cognate3 LLC
Priority date: 2020-09-09
Filing date: 2021-09-09
Publication date: 2023-08-08

Abstract

Recombinant adeno-associated virus (rAAV) compositions for expressing antibodies (e.g., anti-complement component 5 (C5) antibodies) in cells, and methods of treating disorders (e.g., disorders associated with C5 activity (e.g., paroxysmal sleep hemoglobinuria)) with the compositions are provided. Compositions, systems, and methods for preparing rAAV compositions are also provided.

Description

Supported antibodies and uses thereof

RELATED APPLICATIONS

The present application claims priority from U.S. provisional patent application Ser. No. 63/075,898, filed 9/2020, and U.S. provisional patent application Ser. No. 63/179,990, filed 26/2021, 4, the disclosures of which are hereby incorporated by reference in their entireties.

Sequence listing

The present application contains a sequence listing that has been electronically submitted in ASCII format and incorporated herein by reference in its entirety (the ASCII copy was created at 9, 8, 2021, named "404217-HMW-042wo_185407_st25.Txt" and is 252,035 bytes in size).

Background

Therapeutic antibodies represent a potent class of drugs that are highly specific for a target of interest. However, many antibodies require large individual doses and periodic administration to achieve the desired therapeutic effect. This is especially true for high concentration antibody targets found in the serum of patients. For example, anti-complement component 5 (C5) antibodies are used to treat C5 mediated diseases such as Paroxysmal Nocturnal Hemoglobinuria (PNH), neuromyelitis optica (NMOSD) spectrum disorders, and atypical hemolytic uremic syndrome (aHUS), requiring multiple large doses of antibodies due to the high abundance of C5 in serum.

The viral delivery mechanism provides an attractive alternative to traditional antibody therapy, particularly for high concentration antibody targets found in the serum of patients. In particular, a single administration of a viral vector having antibody heavy and light chain expression cassettes makes it possible to produce sustained therapeutic levels of antibodies in the serum of a subject, thus eliminating the need for continuous administration of high doses of antibodies.

Thus, there is a need in the art for improved viral vectors to efficiently and consistently express antibodies in a subject.

Disclosure of Invention

Provided herein are recombinant adeno-associated virus (rAAV) compositions for expressing antibodies (e.g., anti-complement component 5 (C5) antibodies) in cells, and methods of treating disorders (e.g., disorders associated with C5 activity (e.g., paroxysmal sleep hemoglobinuria)) with the compositions. Compositions, systems, and methods for preparing rAAV compositions are also provided.

Accordingly, in one aspect, the present disclosure provides a recombinant adeno-associated virus (rAAV) genome comprising:

(a) A first expression cassette comprising, from 5 'to 3':

a first liver-specific transcription regulatory element,

a first coding sequence encoding a first polypeptide comprising an antibody heavy chain operably linked to a first signal sequence, an

A first polyadenylation sequence; and

(b) A second expression cassette comprising, from 5 'to 3':

a second liver-specific transcription regulatory element,

a second coding sequence encoding a second polypeptide comprising an antibody light chain operably linked to a second signal sequence, an

A second polyadenylation sequence which is complementary to the first polyadenylation sequence,

wherein expression of the first coding sequence and the second coding sequence produces an antibody comprising the antibody heavy chain and the antibody light chain.

In certain embodiments, the first transcription regulatory element and/or the second transcription regulatory element comprises a promoter element selected from the group consisting of: human albumin promoter, human transthyretin (TTR) promoter, human thyroxine-binding globulin (TBG) promoter, human ApoH promoter, human SERPINA1 (hAAT) promoter and liver-specific regulatory modules thereof, such as human ApoE/C-I liver control region (HCR) 1 or 2.

In certain embodiments, the first transcription regulatory element and/or the second transcription regulatory element comprises a promoter element comprising a nucleic acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs 25, 27, 66, 68, 69, 116 and 117.

In certain embodiments, the transcriptional regulatory element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 27. In certain embodiments, the transcriptional regulatory element comprises the nucleotide sequence set forth in SEQ ID NO 27. In certain embodiments, the nucleotide sequence of the transcriptional regulatory element consists of the nucleotide sequence set forth in SEQ ID NO. 27.

In certain embodiments, the transcriptional regulatory element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 67. In certain embodiments, the transcriptional regulatory element comprises the nucleotide sequence set forth in SEQ ID NO. 67. In certain embodiments, the nucleotide sequence of the transcriptional regulatory element consists of the nucleotide sequence set forth in SEQ ID NO: 67.

In certain embodiments, the first expression cassette and/or the second expression cassette further comprise an intron element that is located 5 'relative to the first coding sequence and/or the second coding sequence and 3' relative to the transcriptional regulatory element.

In certain embodiments, the intron element is an exogenous intron element, optionally wherein the exogenous intron element is an SV40 intron element or a mouse adenovirus (MVM) intron element.

In certain embodiments, the SV40 intron elements comprise a nucleotide sequence that is at least 90% identical to the nucleotide sequence depicted in SEQ ID NO. 29. In certain embodiments, the SV40 intron elements comprise the nucleotide sequence depicted in SEQ ID NO. 29. In certain embodiments, the nucleotide sequence of the SV40 intron element consists of the nucleotide sequence depicted in SEQ ID NO. 29.

In certain embodiments, the MVM intron elements comprise a nucleotide sequence at least 90% identical to the nucleotide sequence depicted in SEQ ID NO. 30. In certain embodiments, the MVM intron element comprises the nucleotide sequence depicted in SEQ ID NO. 30. In certain embodiments, the nucleotide sequence of the MVM intron element consists of the nucleotide sequence depicted in SEQ ID NO. 30.

In certain embodiments, the first transcription regulatory element and the second transcription regulatory element are the same.

In certain embodiments, the first transcriptional regulatory element comprises an HCR 1 element, an hAAT promoter, and an SV40 intron element, and the second transcriptional regulatory element comprises a SERPINA1 liver-specific regulatory module, a TTR promoter, and an MVM intron element.

In certain embodiments, the first transcriptional regulatory element comprises the nucleic acid sequence of SEQ ID NO. 50 and the second transcriptional regulatory element comprises the nucleic acid sequence of SEQ ID NO. 43.

In certain embodiments, the first expression cassette and/or the second expression cassette further comprise a polyadenylation sequence 3' relative to the first coding sequence and/or the second coding sequence.

In certain embodiments, the polyadenylation sequence is an exogenous polyadenylation sequence, optionally wherein the exogenous polyadenylation sequence is an SV40 polyadenylation sequence or a Bovine Growth Hormone (BGH) polyadenylation sequence.

In certain embodiments, the SV40 polyadenylation sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence depicted in SEQ ID NO. 31. In certain embodiments, the SV40 polyadenylation sequence comprises the nucleotide sequence shown in SEQ ID NO. 31. In certain embodiments, the nucleotide sequence of the SV40 polyadenylation sequence consists of the nucleotide sequence depicted in SEQ ID NO. 31.

In certain embodiments, the BGH polyadenylation sequence comprises a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 33. In certain embodiments, the BGH polyadenylation sequence comprises the nucleotide sequence set forth in SEQ ID NO. 33. In certain embodiments, the nucleotide sequence of the BGH polyadenylation sequence consists of the nucleotide sequence set forth in SEQ ID NO. 33.

In certain embodiments, the first expression cassette and the second expression cassette comprise the same polyadenylation sequence.

In certain embodiments, the first expression cassette comprises an SV40 polyadenylation sequence. In certain embodiments, the second expression cassette comprises a BGH polyadenylation sequence.

In certain embodiments, the first polyadenylation sequence comprises the nucleic acid sequence of SEQ ID NO. 31 and the second polyadenylation sequence comprises the nucleic acid sequence of SEQ ID NO. 33.

In certain embodiments, the first expression cassette and the second expression cassette are in the same orientation in the rAAV genome. In certain embodiments, the first expression cassette and the second expression cassette are in opposite orientations in the rAAV genome.

In certain embodiments, the first expression cassette and the second expression cassette are in opposite orientations, wherein the first polyadenylation sequence and the second polyadenylation sequence are distally located in the rAAV genome.

In certain embodiments, the rAAV genome further comprises a stuffer sequence interposed between the first transcriptional regulatory element and the second transcriptional regulatory element.

In certain embodiments, the stuffer sequence comprises a β globin polyadenylation sequence. In certain embodiments, the beta globin polyadenylation sequence comprises the nucleic acid sequence of SEQ ID NO: 51.

In certain embodiments, the rAAV genome comprises, from 5 'to 3': (a) A first polyadenylation sequence comprising the nucleic acid sequence of SEQ ID NO. 33; (b) a first coding sequence; (c) A first liver-specific transcriptional regulatory element comprising a nucleic acid sequence of SEQ ID NO. 27; (d) a stuffer sequence comprising the nucleic acid sequence of SEQ ID NO. 51; (e) A second liver-specific transcriptional regulatory element comprising the nucleic acid sequence of SEQ ID NO. 67; (f) a second coding sequence; (g) A second transcribed polyadenylation sequence comprising the nucleic acid sequence of SEQ ID NO. 31.

In certain embodiments, the rAAV genome comprises, from 5 'to 3': reverse complement of the first expression cassette; a filling sequence; and a second expression cassette.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 27, a first coding sequence, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 33; (b) A stuffer sequence comprising a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 67, a second coding sequence, a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 31.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 67, a first coding sequence, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 31; (b) A stuffer sequence comprising a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 27, a second coding sequence, a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 33.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 25, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 26, a first coding sequence, a first polyadenylation sequence; (b) A stuffer sequence comprising a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 119, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 45, a second coding sequence, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 31.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 119, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 45, a first coding sequence, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 31; (b) A stuffer sequence comprising a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 25, a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 26, a second coding sequence, a first polyadenylation sequence.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 27, the first coding sequence, the nucleotide sequence shown in SEQ ID NO. 33; (b) A stuffer sequence comprising the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 67, the second coding sequence, the nucleotide sequence shown in SEQ ID NO. 31.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 67, the first coding sequence, the nucleotide sequence shown in SEQ ID NO. 31; (b) A stuffer sequence comprising the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 27, the second coding sequence, the nucleotide sequence shown in SEQ ID NO. 33.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 25, the nucleotide sequence shown in SEQ ID NO. 26, the first coding sequence, the first polyadenylation sequence; (b) A stuffer sequence comprising the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 119, the nucleotide sequence shown in SEQ ID NO. 45, the second coding sequence, the nucleotide sequence shown in SEQ ID NO. 31.

In certain embodiments, the rAAV genome comprises: (a) A first expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 119, the nucleotide sequence shown in SEQ ID NO. 45, the first coding sequence, the nucleotide sequence shown in SEQ ID NO. 31; (b) A stuffer sequence comprising the nucleotide sequence set forth in SEQ ID No. 51 or an inverse complement thereof; and (c) a second expression cassette comprising, from 5 'to 3': the nucleotide sequence shown in SEQ ID NO. 25, the nucleotide sequence shown in SEQ ID NO. 26, the second coding sequence, the first polyadenylation sequence.

In one aspect, the disclosure provides a rAAV genome comprising a bicistronic expression cassette comprising, from 5 'to 3':

(a) A liver-specific transcriptional regulatory element; a first coding sequence encoding a first polypeptide comprising an antibody heavy chain operably linked to a first signal sequence; a ribosome-hopping sequence encoding a ribosome-hopping peptide; a second coding sequence encoding a second polypeptide comprising an antibody light chain operably linked to a second signal sequence; and polyadenylation sequences, or

(b) A liver-specific transcriptional regulatory element; a second coding sequence encoding a second polypeptide comprising an antibody light chain operably linked to a second signal sequence; a ribosome-hopping sequence encoding a ribosome-hopping peptide; a first coding sequence encoding a first polypeptide comprising an antibody heavy chain operably linked to a first signal sequence; a polyadenylation sequence which is complementary to the polyadenylation sequence,

wherein expression of the bicistronic expression cassette results in an antibody comprising the antibody heavy chain and the antibody light chain.

In certain embodiments, the transcriptional regulatory element comprises a promoter element selected from the group consisting of: human albumin promoter, human transthyretin (TTR) promoter, human thyroxine-binding globulin (TBG) promoter, human ApoH promoter, human SERPINA1 (hAAT) promoter and liver-specific regulatory modules thereof, such as human ApoE/C-I liver control region (HCR) 1 or 2.

In certain embodiments, the transcriptional regulatory element comprises a promoter element comprising a nucleic acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs 25, 27, 66, 68, 69, 116 and 117.

In certain embodiments, the bicistronic expression cassette further comprises an intron element 5 'relative to the first coding sequence and/or the second coding sequence and 3' relative to the transcriptional regulatory element.

In certain embodiments, the transcriptional regulatory element comprises:

a) HCR 1 element, hAAT promoter and SV40 intron element; or (b)

b) SERPINA1 liver-specific regulatory modules, TTR promoter, and MVM intron elements.

In certain embodiments, the transcriptional regulatory element comprises the nucleic acid sequence of SEQ ID NO. 50 or the nucleic acid sequence of SEQ ID NO. 43.

In certain embodiments, the first signal sequence and/or the second signal sequence is a naturally occurring signal sequence. In certain embodiments, the first signal sequence and/or the second signal sequence is an antibody signal sequence, optionally a human IgG2 or IgK signal sequence. In certain embodiments, the first signal sequence and/or the second signal sequence is a non-naturally occurring signal sequence. In certain embodiments, the first signal sequence and/or the second signal sequence comprises the amino acid sequence of SEQ ID NO. 80. In certain embodiments, the first signal sequence and/or the second signal sequence comprises the amino acid sequence of SEQ ID NO. 81. In certain embodiments, the first signal sequence comprises the amino acid sequence of SEQ ID NO. 80 and the second signal sequence comprises the amino acid sequence of SEQ ID NO. 81. In certain embodiments, the first coding sequence and/or the second coding sequence comprises any of the nucleic acid sequences set forth in SEQ ID NO. 23, 96, 102 or 108. In certain embodiments, the first coding sequence and/or the second coding sequence comprises any of the nucleic acid sequences set forth in SEQ ID NO. 24, 99, 105, 111 or 130. In certain embodiments, the first coding sequence comprises any of the nucleic acid sequences set forth in SEQ ID NO. 23, 96, 102 or 108 and the second coding sequence comprises any of the nucleic acid sequences set forth in SEQ ID NO. 24, 99, 105, 111 or 130.

In certain embodiments, the antibody specifically binds to complement C5.

In certain embodiments, the antibody heavy chain comprises the amino acid sequence of SEQ ID NO. 64. In certain embodiments, the antibody heavy chain comprises the amino acid sequence of SEQ ID NO. 82. In certain embodiments, the antibody light chain comprises the amino acid sequence of SEQ ID NO. 77.

In certain embodiments, the first coding sequence and/or the second coding sequence have been optimized for expression in human cells.

In certain embodiments, the first coding sequence comprises any of the nucleic acid sequences set forth in SEQ ID NOs 52, 113, 114 or 115.

In certain embodiments, the first coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NO 83, 94, 95, 101 or 107.

In certain embodiments, the second coding sequence comprises any of the nucleic acid sequences set forth in SEQ ID NO 53, 98, 104, 110 or 131.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 53.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO: 63.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 98.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 99.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 100.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 104.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 105.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 106.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 110.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 111.

In certain embodiments, the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114 and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 112.

In certain embodiments, the rAAV genome is a single-stranded rAAV genome.

In certain embodiments, the rAAV genome is a self-complementary rAAV genome.

In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 84. In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 85. In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 86. In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 87.

In certain embodiments, the rAAV genome further comprises a 5 'inverted terminal repeat (5' itr) nucleotide sequence 5 'relative to the first polyadenylation sequence and a 3' inverted terminal repeat (3 'itr) nucleotide sequence 3' relative to the second polyadenylation sequence.

In certain embodiments, the 5'ITR nucleotide sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleotide sequence set forth in SEQ ID NO:14, and/or the 3' ITR nucleotide sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleotide sequence set forth in SEQ ID NO: 18.

In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 88. In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 89. In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 90. In certain embodiments, the rAAV genome comprises the nucleic acid sequence of SEQ ID NO. 91.

In another aspect, the present disclosure provides a recombinant adeno-associated virus (rAAV) comprising:

(a) AAV capsids, the AAV capsids comprising AAV capsid proteins; and

(b) The rAAV genome of any preceding embodiment.

In certain embodiments, the capsid protein is selected from the group consisting of: AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, and AAV9.

In certain embodiments, the AAV capsid protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of amino acids 203-736 of SEQ ID NO. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G.

In some embodiments of the present invention, in some embodiments,

(a) The amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G and the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G;

(b) Amino acid corresponding to amino acid 296 of SEQ ID NO. 16 in the capsid protein is H, amino acid corresponding to amino acid 464 of SEQ ID NO. 16 in the capsid protein is N, amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and amino acid corresponding to amino acid 681 of SEQ ID NO. 16 in the capsid protein is M;

(c) The amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R;

(d) The amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A and the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; or alternatively

(e) The amino acid corresponding to amino acid 501 of SEQ ID NO. 16 in the capsid protein is I, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 706 of SEQ ID NO. 16 in the capsid protein is C.

In certain embodiments, the capsid protein comprises the amino acid sequence of amino acids 203-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the AAV capsid protein comprises an amino acid sequence at least 95% identical to the amino acid sequence of amino acids 138-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 151 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 160 of SEQ ID NO. 16 is D; the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G.

In some embodiments of the present invention, in some embodiments,

In certain embodiments, the capsid protein comprises the amino acid sequence of amino acids 138-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the AAV capsid protein comprises an amino acid sequence at least 95% identical to the amino acid sequence of amino acids 1-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 2 of SEQ ID NO. 16 is T; the amino acid in the capsid protein corresponding to amino acid 65 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 68 of SEQ ID NO. 16 is V; the amino acid in the capsid protein corresponding to amino acid 77 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 119 of SEQ ID NO. 16 is L; the amino acid in the capsid protein corresponding to amino acid 151 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 160 of SEQ ID NO. 16 is D; the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G.

In some embodiments of the present invention, in some embodiments,

(a) The amino acid in the capsid protein corresponding to amino acid 2 of SEQ ID NO. 16 is T and the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q;

(b) The amino acid in the capsid protein corresponding to amino acid 65 of SEQ ID NO. 16 is I and the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is Y;

(c) The amino acid in the capsid protein corresponding to amino acid 77 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K;

(d) The amino acid in the capsid protein corresponding to amino acid 119 of SEQ ID NO. 16 is L and the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S;

(e) The amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G and the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G;

(f) Amino acid corresponding to amino acid 296 of SEQ ID NO. 16 in the capsid protein is H, amino acid corresponding to amino acid 464 of SEQ ID NO. 16 in the capsid protein is N, amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and amino acid corresponding to amino acid 681 of SEQ ID NO. 16 in the capsid protein is M;

(g) The amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R;

(h) The amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A and the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; or alternatively

(i) The amino acid corresponding to amino acid 501 of SEQ ID NO. 16 in the capsid protein is I, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 706 of SEQ ID NO. 16 in the capsid protein is C.

In certain embodiments, the capsid protein comprises the amino acid sequence of amino acids 1-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In one aspect, the present disclosure provides a polynucleotide comprising the nucleic acid sequence set forth in SEQ ID NOS.85-93.

In one aspect, the present disclosure provides a pharmaceutical composition comprising a rAAV as described above or a polynucleotide as described above.

In one aspect, the present disclosure provides a packaging system for preparing a rAAV, wherein the packaging system comprises:

(a) A first nucleotide sequence encoding one or more AAV Rep proteins;

(b) A second nucleotide sequence encoding a capsid protein of a rAAV as described above; and

(c) A third nucleotide sequence comprising a rAAV genomic sequence of a rAAV as described above.

In certain embodiments, the packaging system comprises a first vector comprising a first nucleotide sequence and a second nucleotide sequence, and a second vector comprising a third nucleotide sequence.

In certain embodiments, the packaging system further comprises a fourth nucleotide sequence comprising one or more helper virus genes. In certain embodiments, the fourth nucleotide sequence is included within the third vector. In certain embodiments, the fourth nucleotide sequence comprises one or more genes from a virus selected from the group consisting of: adenovirus, herpes virus, vaccinia virus and Cytomegalovirus (CMV).

In certain embodiments, the first vector, the second vector, and/or the third vector are plasmids.

In one aspect, the present disclosure provides a method for recombinantly producing an rAAV comprising introducing into a cell a packaging system described above under conditions whereby the rAAV is produced.

In another aspect, the disclosure provides a rAAV as described herein, a pharmaceutical composition as described herein, or a polynucleotide as described herein for use as a medicament.

In another aspect, the disclosure provides a rAAV as described above, a pharmaceutical composition as described above, or a polynucleotide as described above for use in treating a complement C5-associated disease.

In another aspect, the present disclosure provides a rAAV as described above, a pharmaceutical composition as described above, or a polynucleotide as described above for use in a method of treating a subject having a complement C5-associated disease, the method comprising administering to the subject an effective amount of the rAAV, pharmaceutical composition, or polynucleotide.

In one aspect, the present disclosure provides a method of producing an antibody in a subject, the method comprising administering to the subject a pharmaceutical composition described above.

In certain embodiments, the pharmaceutical composition is administered intravenously.

In one aspect, the present disclosure provides a method of treating a complement C5-associated disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a rAAV described above, a pharmaceutical composition described above, or a polynucleotide described above.

In certain embodiments, the complement C5-associated disease is selected from the group consisting of: geographic Atrophy (GA), guillain-Barre syndrome (Guillain-Barre syndrome), myasthenia gravis, systemic Lupus Erythematosus (SLE) nephritis, proliferative nephritis, asthma, rheumatoid arthritis, sepsis, paroxysmal sleep hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), and age-related macular degeneration (AMD).

In certain embodiments, the rAAV, pharmaceutical composition, or polynucleotide is administered intravenously.

Drawings

FIG. 1 depicts a vector diagram of expression cassettes for rAAV vectors C5Ab01, C5Ab02, C5Ab03, and C5Ab 04.

Fig. 2A-2I depict graphs showing the concentration of anti-C5 antibodies in serum of NOD SCID mice receiving anti-C5 antibody expression vectors (C5 Ab02, C5Ab03, and C5Ab 04) packaged in AAVHSC13, AAVHSC15, or AAVHSC17 capsids. FIG. 2A depicts a graph showing anti-C5 antibody concentration in serum of mice receiving 1e13 vg/kg dose of vector C5Ab04 packaged in AAVHSC13 or AAVHSC17 capsids. Data from male and female mice were isolated and multiple serum samples were collected over a 23 week period. Fig. 2B depicts a graph showing the results in fig. 2A, with the Y-axis on a logarithmic scale. FIG. 2C depicts a graph showing anti-C5 antibody concentration in serum of mice receiving a 1e13 vg/kg dose of vector C5Ab02 packaged in AAVHSC17 capsid. Data from male and female mice were isolated and multiple serum samples were collected over a 16 week period. Fig. 2D depicts a graph showing the results in fig. 2C, with the Y-axis on a logarithmic scale. FIG. 2E depicts a graph showing anti-C5 antibody concentrations in serum of mice receiving 1E13 vg/kg doses of vector C5Ab02, C5Ab03, or C5Ab04 packaged in AAVHSC15 or AAVHSC17 capsids, respectively. Data from male mice are shown and multiple serum samples are collected over a 16 week period. Fig. 2F shows the results in fig. 2E depicted in a line graph format, and fig. 2G shows the results in fig. 2E depicted in a line graph format, with the Y-axis being on a logarithmic scale. FIG. 2H depicts a graph showing the concentration of anti-C5 antibodies in serum of mice receiving 5e11 vg/kg, 5e12 vg/kg, 1.4e13 vg/kg, 4.4e13 vg/kg, and 1.8e14 vg/kg of 5 doses of vector C5Ab04 packaged in AAVHSC17 capsids. Data from male mice are shown and multiple serum samples are collected over a 13 week period. Fig. 2I depicts a graph showing the results in fig. 2H, with the Y-axis on a logarithmic scale.

FIGS. 3A-3C depict graphs showing anti-C5 antibody concentration in serum of NOD SCID male mice receiving an anti-C5 antibody expression vector. The data were derived from fig. 2 above and ranked to compare vectors C5Ab02, C5Ab03 or C5Ab04 packaged in AAVHSC13, AAVHSC15 or AAVHSC17 capsids. FIG. 3A depicts a graph showing anti-C5 antibody concentration in serum of mice receiving a 1e13 vg/kg dose of vector C5Ab04 packaged in AAVHSC13 capsid. Fig. 3B depicts a graph showing anti-C5 antibody concentration in serum of mice that received a 1e13 vg/kg dose of vector C5Ab02, C5Ab03, or C5Ab04 packaged in AAVHSC15 capsids. Fig. 3C depicts a graph showing anti-C5 antibody concentration in serum of mice that received a 1e13 vg/kg dose of vector C5Ab02, C5Ab03, or C5Ab04 packaged in AAVHSC17 capsids.

Fig. 4A depicts a graph comparing predicted anti-C5 antibody concentrations in PNH patients receiving anti-C5 antibody eculizumab (eculizumab) and Lei Fuli bead mab (ravulizumab) long-term maintenance therapy with anti-C5 antibody concentrations measured in NOD SCID male and female mice using AAVHSC13 or AAVHSC17 capsids (data from fig. 2B). Fig. 4B depicts a graph comparing predicted anti-C5 antibody concentrations in PNH patients receiving anti-C5 antibodies eculizumab and Lei Fuli beadmab to anti-C5 antibody concentrations measured in NOD SCID and HuLiv mice using AAVHSC17 capsids (representative data from fig. 2A, 2I and 8B).

Figure 5 depicts a graph of% hemolysis of activated sheep Red Blood Cells (RBCs) at various concentrations of anti-C5 antibodies in an ex vivo hemolysis assay. anti-C5 control antibodies were compared to sera obtained from mice treated with AAVHSC 13-packaged C5Ab04 and AAVHSC 17-packaged C5Ab 04.

Fig. 6A depicts a plot of serum antibody concentration and fig. 6B depicts a plot of% hemolysis of activated sheep RBCs in an ex vivo hemolysis assay. Negative control mouse serum was compared to serum obtained from mice treated with AAVHSC 13-packaged C5Ab04 and AAVHSC 17-packaged C5Ab04 at 1 week, 3 weeks, 5 weeks, 7 weeks, and 9 weeks after each administration at a dose of 1e13 vgs/kg. Fig. 6C depicts a graph showing the results in fig. 6B, wherein% hemolysis is determined from serum samples obtained 19 weeks after administration and presented as a line graph. Fig. 6D depicts a graph of% hemolysis of activated sheep RBCs in an ex vivo hemolysis assay using serum obtained from mice treated with AAVHSC17 packaged C5Ab02 at a dose of 1e13 vg/kg. Fig. 6E depicts a graph of% hemolysis of activated sheep RBCs in an ex vivo hemolysis assay performed using serum obtained from mice treated with AAVHSC15 or AAVHSC17 packaged C5Ab02, C5Ab03, or C5Ab04, each at a dose of 1E13 vg/kg. FIG. 6F depicts a graph of% hemolysis of activated sheep RBC in an ex vivo hemolysis assay performed using serum obtained from mice treated with AAVHSC17 packaged C5Ab04 at doses of 5e11 vg/kg, 5e12 vg/kg, 1.4e13 vg/kg, 4.4e13 vg/kg, and 1.8e14 vg/kg. In fig. 6A-6F, data from male and female mice are isolated, and data in fig. 6E and 6F are from male mice.

Fig. 7A-7B depict graphs comparing: human C5 levels in serum of FRGKO humanized liver mice (hereinafter HuLiv, yecuris) and human C5 levels found in human serum in the background of C57Bl/6 (FRGC 57) or NOD (FRGNOD) (fig. 7A); and mouse C5 levels in serum of HuLiv mice in C57Bl6 or NOD background (fig. 7B). Frgc57_donor a represents a HuLiv mouse (n=3) with hepatocytes from patient donor a. Frgc57_donor B represents a HuLiv mouse (n=3) with hepatocytes from patient donor B. Nod_donor a represents HuLiv mice (n=9) from homologous NOD mice with hepatocytes from patient donor a.

FIG. 8A depicts a graph of serum antibody concentration of anti-C5 antibody in HuLiv mice of C5Ab04 packaged in AAVHSC17 capsids administered at a dose of 100 μg anti-C5 antibody (bio-mimetic pharmaceuticals) or 1e13 vg/kg or 1e14 vg/kg, in each case 0 week, 1 week, 3 weeks and 5 weeks after administration. In fig. 8A, PB indicates before blood drawing. Fig. 8B shows the data depicted in fig. 8A, wherein serum antibody concentrations were determined at week 11 after administration, presented as a line graph. FIG. 8C depicts a graph of% hemolysis of activated sheep RBC in an ex vivo hemolysis assay using serum obtained from mice administered 100 μg of anti-C5 antibody (bio-mimetic pharmaceutical) or a dose of 1e13 vg s/kg or 1e14 vg s/kg of C5Ab04 packaged in AAVHSC17 capsid. FIG. 8D depicts a graph showing the levels of mouse C5 detected in serum obtained from mice administered a single dose of 100 μg of anti-C5 antibody (biomimetic) or 1e13 vg/kg or 1e14 vg/kg of C5Ab04 packaged in AAVHSC17 capsid. Fig. 9A-9B depict western blots of human C5 levels in media of primary human and mouse hepatocytes transduced with C5Ab02, C5Ab03, or C5Ab04 packaged in AAVHSC15 or AAVHSC17 capsids (fig. 9A) and human IgG ELISA data (9B).

Detailed Description

Provided herein are rAAV genomes and rAAV for expressing antibodies (e.g., anti-C5 antibodies) in cells (e.g., hepatocytes), and methods of using the rAAV genomes and rAAV to treat disorders (e.g., disorders associated with C5 activity (e.g., paroxysmal sleep hemoglobinuria)). Nucleic acids, vectors, packaging systems, and methods for making rAAV are also provided.

I. Definition of the definition

As used herein, the term "recombinant adeno-associated virus" or "rAAV" refers to an AAV that includes a genome lacking functional rep and cap genes.

As used herein, the term "rAAV genome" refers to a nucleic acid molecule (e.g., DNA and/or RNA) that includes the genomic sequence of a rAAV. Those of skill in the art will appreciate that when the rAAV genome includes a transgene (e.g., an antibody heavy or light chain coding sequence operably linked to transcriptional regulatory elements), the rAAV genome may be in a sense or antisense orientation relative to the direction of transcription of the transgene.

As used herein, the term "AAV capsid protein" refers to an AAV VP1, VP2 or VP3 capsid protein.

As used herein, the "percent identity" between two nucleotide sequences or between two amino acid sequences is calculated by multiplying the number of matches between aligned pairs of sequences by 100 and dividing by the length of the aligned regions (including internal gaps). Identity scores only count perfect matches and do not take into account the degree of similarity of amino acids to each other. It should be noted that only internal gaps are included in length, and no gaps at the ends of the sequence are included.

As used herein, the term "coding sequence" refers to the portion of complementary DNA (cDNA) that encodes a polypeptide, beginning at the start codon and ending at the stop codon. Genes may have one or more coding sequences due to alternative splicing, alternative translation initiation, and changes within the population. The coding sequence may be wild-type, silenced, or intronic. Exemplary anti-C5 heavy chain coding sequences are shown in SEQ ID NOs 52 and 83. An exemplary anti-C5 light chain coding sequence is shown in SEQ ID NO. 53. The coding sequence may be codon optimized. Codon optimization may be performed to enhance expression of the coding sequence in a desired host cell (e.g., a human cell). Exemplary codon optimized anti-C5 heavy chain coding sequences are shown in SEQ ID NOs 94, 95, 101, 107, 113, 114 and 115. Exemplary codon optimized anti-C5 light chain coding sequences are shown in SEQ ID NOs 98, 104, 110 or 131.

In certain embodiments, two or more coding sequences (e.g., an antibody heavy chain coding sequence and an antibody light chain coding sequence) may be separated by a nucleotide sequence (e.g., a 2A peptide ribosome-hopping element) that encodes a peptide cleavage sequence. Exemplary 2A peptide cleavage sequences are shown in SEQ ID NO 28 or 125 (T2A peptide cleavage sequence) or 128 (P2A peptide cleavage sequence). The 2A peptide cleavage sequence may also include a furin cleavage sequence and a linker. Exemplary 2A peptide cleavage sequences having furin cleavage sequences and linkers are shown in SEQ ID NO:127 or 129.

As used herein, the term "polyadenylation sequence" refers to a DNA sequence that, when transcribed into RNA, constitutes a polyadenylation signal sequence. Polyadenylation sequences may be native or exogenous. The exogenous polyadenylation sequence may be a mammalian or viral polyadenylation sequence (e.g., bovine growth hormone polyadenylation sequence or SV40 polyadenylation sequence).

As used herein, the term "intron element" refers to a cis-acting nucleotide sequence, e.g., a DNA sequence, that modulates (e.g., controls, increases, or decreases) expression of a transgene. In certain embodiments, the intron element is a modified intron, e.g., a synthetic intron sequence. In certain embodiments, the intron element is an exogenous intron element and is derived from an intron for which the regulatable transgene is exogenous. In certain embodiments, the intron element comprises a modified splice acceptor and/or splice donor, resulting in more robust splice activity. While not wishing to be bound by theory, it is hypothesized that introns may increase transgene expression, for example, by reducing transcriptional silencing and enhancing mRNA export from the nucleus to the cytoplasm. The skilled artisan will appreciate that synthetic intron sequences may be designed to mediate RNA splicing by introducing any consensus splice motif known in the art (e.g., in Sibley et al, (2016) Nature comment: genetics, 17,407-21, incorporated herein by reference in its entirety). Exemplary intron sequences are provided in Lu et al, (2013) molecular therapy 21 (5): 954-63 and Lu et al, (2017) human Gene therapy 28 (1): 125-34, which are incorporated herein by reference in their entirety.

As used herein, the term "silenced change" refers to a change in the coding sequence of a gene (e.g., by nucleotide substitution) without changing the amino acid sequence of the polypeptide encoded by the coding sequence or the coding sequence inserted through a filler. Such silent alterations are advantageous in that they may increase the translational efficiency of the coding sequence and/or prevent recombination with the corresponding sequence of the endogenous gene when the coding sequence is transduced into a cell.

As used herein, the term "transcriptional regulatory element" or "TRE" refers to a cis-acting nucleotide sequence, e.g., a DNA sequence, that modulates (e.g., controls, increases, or decreases) transcription of an operably linked nucleotide sequence by an RNA polymerase to form an RNA molecule. TRE relies on one or more trans-acting molecules, such as transcription factors, to mediate transcription. Thus, a TRE can modulate transcription in different ways when it comes into contact with different trans-acting molecules (e.g., when it is located in different types of cells). The TRE may comprise one or more promoter elements and/or enhancer elements. Those skilled in the art will appreciate that promoter and enhancer elements in a gene may be located close together, and that the term "promoter" may refer to sequences that include promoter elements and enhancer elements. Thus, the term "promoter" does not exclude enhancer elements in a sequence. The promoter and enhancer elements need not be derived from the same gene or species, and the sequence of each promoter or enhancer element may be identical or substantially identical to the corresponding endogenous sequence in the genome.

As used herein, the term "operably linked" is used to describe a linkage between a TRE and a coding sequence to be transcribed. Typically, gene expression is under the control of a TRE that includes one or more promoter and/or enhancer elements. A coding sequence is "operably linked" to a TRE if the transcription of the coding sequence is controlled or effected by the TRE. The promoter and enhancer elements of the TRE may be any orientation and/or distance from the coding sequence so long as the desired transcriptional activity is obtained. In certain embodiments, the TRE is located upstream of the coding sequence.

In the present disclosure, a nucleotide position in an antibody coding sequence (e.g., an antibody heavy chain coding sequence or an antibody light chain coding sequence) is specified relative to a first nucleotide of a start codon. The first nucleotide of the start codon is position 1; nucleotides 5' relative to the first nucleotide of the start codon have a negative number; the nucleotide at 3' relative to the first nucleotide of the start codon has a positive number.

As used herein, the term "expression cassette" refers to a polynucleotide sequence that includes, from 5 'to 3', transcription Regulatory Elements (TREs), coding sequences encoding polypeptides, and polyadenylation sequences. In certain embodiments, an intron is present between the TRE and the coding sequence. In certain embodiments, the coding sequence encodes an antibody heavy chain or an antibody light chain.

As used herein, the term "effective amount" in the context of administration of AAV to a subject refers to the amount of AAV that achieves a desired prophylactic or therapeutic effect.

As used herein, the term "about" or "approximately" when referring to measurable values such as the expression level of an antibody (e.g., antibody heavy and antibody light chains) encompasses variations of ±20% or ±10%, of ±5%, of ±1% or of ±0.1% of a given value or range, as appropriate for performing the methods disclosed herein.

Adeno-associated virus compositions

In one aspect, provided herein are novel rAAV genomes comprising a Transcription Regulatory Element (TRE) operably linked to at least a portion of an antibody coding sequence (e.g., an anti-C5 antibody heavy chain coding sequence and/or an anti-C5 antibody light chain coding sequence). The rAAV genomes provided herein can be used for extrachromosomal expression of antibodies in cells comprising the rAAV genomes.

The rAAV genome can be used to express an antibody in any mammalian cell (e.g., a human cell). Thus, the TRE may be active in any mammalian cell (e.g., human cell). In certain embodiments, the TRE is active in a wide range of human cells. Such TREs can include constitutive promoter and/or enhancer elements comprising a Cytomegalovirus (CMV) promoter/enhancer (e.g., comprising a sequence identical to SEQ ID NO:54, 55, or 56, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical), SV40 promoter, chicken ACTB promoter (e.g., comprising a nucleotide sequence identical to SEQ ID NO:47 or 57 at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical nucleotide sequence), jeT promoter (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical nucleotide sequence to SEQ ID NO: 58), smCBA promoter (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical nucleotide sequence to SEQ ID NO: 59), human elongation factor 1 α (1 α) promoter (e.g., comprising at least 80%, 81%, 82%, 83%, 84%, 85%, 86% or 100% amino acid sequence of SEQ ID NO:39 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical), a mouse adenovirus (MVM) intron comprising a transcription factor binding site (e.g., comprising a nucleotide sequence identical to SEQ ID NO:30 or 61, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical nucleotide sequence), a human phosphoglycerate kinase (PGK 1) promoter, a human ubiquitin C (Ubc) promoter, a human beta actin promoter, a human neuron-specific enolase (ENO 2) promoter, a human beta-Glucuronidase (GUSB) promoter, a rabbit beta-globin element (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical nucleotide sequence to SEQ ID NO: 41), a human calmodulin 1 (CALM 1) promoter (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 92%, 94%, 96%, 97%, 96%, 99% or 100% identical to SEQ ID NO: 44%, or a methyl 2% promoter). Any of these TREs may be combined in any order to drive efficient transcription. For example, the rAAV genome can include a CMV enhancer, a CBA promoter, and a splice acceptor from exon 3 of the rabbit β -globin gene, collectively referred to as the CAG promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 42). For example, the rAAV genome can include a hybrid of a CMV enhancer and a CBA promoter, followed by a splice donor and splice acceptor, collectively referred to as CASI promoter regions (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:48 or 65).

Alternatively, the TRE may be a tissue specific TRE, i.e. it is active in a specific tissue and/or organ. The tissue-specific TRE comprises one or more tissue-specific promoter and/or enhancer elements, and optionally one or more constitutive promoter and/or enhancer elements. Those of skill in the art will appreciate that tissue-specific promoter and/or enhancer elements may be isolated from genes specifically expressed in tissue by methods well known in the art.

In certain embodiments, the TRE is liver-specific (e.g., hepatocyte-specific). An exemplary liver-specific TRE may comprise one or more elements selected from the group consisting of: human albumin promoter, human transthyretin (TTR) promoter (e.g., comprising a sequence identical to SEQ ID NO:66 at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical), human APOE/C-I liver control region (HCR) 1 (e.g., comprising a nucleotide sequence identical to SEQ ID NO:25 or 68, or a liver-specific regulatory module thereof (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical), a human APOH promoter, and a human SERPINA1 (hAAT) promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:71, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%). In certain embodiments, the hAAT promoter region comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO. 72. In certain embodiments, the liver-specific TRE comprises a TBG SERPINA7 promoter as described in Yan et al (Gene 2016) 506,289-294) (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 116). In certain embodiments, the liver-specific TRE comprises a TBG SERPINA7 promoter (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 117) as described in Hayashi et al (molecular endocrine (1993) 7 (8), 1049-1060). In certain embodiments, the liver-specific TRE comprises the hAAT SERPINA1 promoter (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 118) as described in Hafenrich et al (blood (1994) 84 (10), 3394-3404). In certain embodiments, the liver-specific TRE comprises a TTR promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 119) as described in Costa et al (molecular and cellular biology (1988) 8 (1), 81-90). In certain embodiments, the liver-specific TRE comprises an ApoA2 promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 120) as described in Kan et al (nucleic acids research (1999) 27 (4), 1104-1117). In certain embodiments, the liver-specific TRE comprises an albumin promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID No. 121) as described in Tang et al (biomedical report (2017) 6,627-632). In certain embodiments, the liver-specific TRE comprises a modified fibrinogen promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID No. 122) as described in kyrelation-Moore et al (molecular therapy (2016) 3,16006). In certain embodiments, the liver-specific TRE comprises a minimal human APOE/C-I liver control region (HCR) 1 promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 123) as described in Dang et al (journal of biochemistry (1995) 270 (38), 22557-85). In certain embodiments, the liver-specific TRE comprises a human APOE/C-I liver control region (HCR) 2 promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 124) as described in Allan et al (journal of biochemistry (1995) 270 (44), 26278-81). Further liver-specific promoter elements are disclosed in WO 2009/130208 and Kramer et al (molecular therapy (2003) 7,375-385), which are incorporated herein by reference in their entirety.

In certain embodiments, the rAAV genome comprises two or more TREs, optionally comprising at least one of the TREs disclosed above. Those skilled in the art will appreciate that any of these TREs may be combined in any order, and that the combination of a constitutive TRE and a tissue-specific TRE may drive efficient and tissue-specific transcription. For example, in certain embodiments, the rAAV genome comprises human HCR1 (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:25, 68, or 123) and a human EF-1 a promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39), optionally wherein the human HCR1 is 5' relative to the human EF-1 a promoter. In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence nucleotide set forth in SEQ ID NO. 60.

Similarly, a combination of two or more tissue-specific TREs can drive efficient and tissue-specific transcription. For example, in certain embodiments, the rAAV genome comprises human HCR1 (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:25, 68, or 123) and an hAAT promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 26), optionally wherein the human HCR1 is 5' relative to the hAAT promoter. In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence set forth in SEQ ID NO. 27. In certain embodiments, the rAAV genome comprises a human HCR1 (e.g., comprising the nucleotide sequence set forth in SEQ ID NO: 25) and an hAAT promoter (e.g., comprising the nucleotide sequence set forth in SEQ ID NO: 26), optionally wherein the human HCR1 is at 5' relative to the hAAT promoter. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 27.

In certain embodiments, the rAAV genome comprises a hAAT promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 71) and a human TTR promoter (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66), optionally wherein the liver-specific regulatory module is 5' relative to the human TTR promoter. In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence set forth in SEQ ID NO. 67. In certain embodiments, the rAAV genome comprises a liver-specific regulatory module comprising an hAAT promoter (e.g., comprising the nucleotide sequence set forth in SEQ ID NO: 71) and a human TTR promoter (e.g., comprising the nucleotide sequence set forth in SEQ ID NO: 66), optionally wherein the liver-specific regulatory module is 5' relative to the human TTR promoter. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 67.

In certain embodiments, the rAAV genome further comprises an intron element 5' relative to at least a portion of an antibody coding sequence. Such intron elements may increase transgene expression, for example, by reducing transcriptional silencing and enhancing export of mRNA from the nucleus to the cytoplasm. In certain embodiments, the rAAV genome comprises, from 5 'to 3': TRE, intron elements, and at least a portion of an antibody coding sequence.

The intronic elements may comprise at least a portion of a natural intronic sequence of an immunoglobulin gene, or the intronic elements may be exogenous intronic elements (e.g., at least comprise intronic sequences from different species or different genes from the same species, and/or synthetic intronic sequences). In certain embodiments, the intronic element is an exogenous intronic element comprising at least a portion of an intronic sequence from a different species. In certain embodiments, the intronic element is an exogenous intronic element comprising at least a portion of an intronic sequence of a different gene from the same species. In certain embodiments, the intron element is an exogenous intron element comprising a synthetic intron sequence. In certain embodiments, the intronic elements are exogenous intronic elements comprising at least intronic sequences from different species or a combination of different genes from the same species and/or synthetic intronic sequences.

The skilled artisan will appreciate that intronic elements may be designed to mediate RNA splicing by introducing any consensus splice motif known in the art (e.g., in Sibley et al, (2016) Nature comment: genetics, 17,407-21, incorporated herein by reference in its entirety). Exemplary intron sequences are provided in Lu et al, (2013) molecular therapy 21 (5): 954-63 and Lu et al, (2017) human Gene therapy 28 (1): 125-34, which are incorporated herein by reference in their entirety.

In certain embodiments, the rAAV genome comprises exogenous intron elements. In certain embodiments, a rAAV comprises an SV40 intron element (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 29), a mouse picovirus (MVM) intron (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:30 or 61), or a synthetic intron (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45). In certain embodiments, the rAAV genome comprises an SV40 intron element (e.g., comprising the nucleotide sequence depicted in SEQ ID NO: 29) or a mouse picornavirus (MVM) intron element (e.g., comprising the nucleotide sequences depicted in SEQ ID NO:30 and 61) or a synthetic intron (e.g., comprising the nucleotide sequence depicted in SEQ ID NO: 45).

In certain embodiments, the rAAV genome comprises, from 5 'to 3': TRE and intron elements. In certain embodiments, the combined TRE and intron element has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID No. 43 or 50. In certain embodiments, the combined TRE and intron elements comprise the nucleotide sequence of SEQ ID NO. 43 or 50. In certain embodiments, the combined TRE and intron elements consist of the nucleotide sequence of SEQ ID NO. 43 or 50.

In certain embodiments, a rAAV genome disclosed herein further comprises a transcription terminator (e.g., a polyadenylation sequence). In certain embodiments, the transcription terminator is 3' relative to at least a portion of the antibody coding sequence. The transcription terminator may be any sequence effective to terminate transcription, and those skilled in the art will appreciate that such sequences may be isolated from any gene expressed in a cell in which transcription of at least a portion of the antibody coding sequence is desired. In certain embodiments, the transcription terminator comprises a polyadenylation sequence. In certain embodiments, the polyadenylation sequence is identical or substantially identical to the endogenous polyadenylation sequence of the immunoglobulin gene. In certain embodiments, the polyadenylation sequence is an exogenous polyadenylation sequence. In certain embodiments, the polyadenylation sequence is an SV40 polyadenylation sequence (e.g., comprising a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to, or complementary to, SEQ ID NO:31, 34, or 35). In certain embodiments, the polyadenylation sequence comprises the nucleotide sequence shown in SEQ ID NO. 31. In certain embodiments, the polyadenylation sequence consists of the nucleotide sequence set forth in SEQ ID NO. 31. In certain embodiments, the polyadenylation sequence is a Bovine Growth Hormone (BGH) polyadenylation sequence (e.g., comprising a nucleotide sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to, or complementary to, SEQ ID NO: 33). In certain embodiments, the polyadenylation sequence comprises the nucleotide sequence set forth in SEQ ID NO. 32. In certain embodiments, the polyadenylation sequence consists of the nucleotide sequence set forth in SEQ ID NO. 32.

In certain embodiments, the rAAV genome comprises, from 5 'to 3': TRE, intron elements, at least a portion of an antibody coding sequence, and a polyadenylation sequence. In certain embodiments, the TRE has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with any one of SEQ ID NOs 25-27, 36, 39, 44, 46-49, 54-60 or 65-72; the intron element has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID No. 29, 30 or 61; at least a portion of the antibody coding sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NOs 52, 53, 62, 63, 83, 94, 95, 97, 98, 100, 101, 103, 104, 106, 107, 109, 110, 112, 113, 114, 115, 131 and 132; and/or the polyadenylation sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any of SEQ ID NOs 31, 33, 34 or 35.

In certain embodiments, the TRE comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 25-27, 36, 39, 42, 44, 46-49, 54-60 and 65-72; the intron elements include a nucleotide sequence selected from the group consisting of SEQ ID NOs 29, 30 and 61; at least a portion of the antibody coding sequence comprises the nucleotide sequence set forth in SEQ ID NO. 52; and/or the polyadenylation sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 31, 33, 34 and 35.

In certain embodiments, the TRE comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 25-27, 36, 39, 42, 44, 46-49, 54-60 and 65-72; the intron elements include a nucleotide sequence selected from the group consisting of SEQ ID NOs 29, 30 and 61; at least a portion of the antibody coding sequence comprises the nucleotide sequence set forth in SEQ ID NO. 53; and/or the polyadenylation sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 31, 33, 34 and 35.

In certain embodiments, the TRE comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 25-27, 36, 39, 42, 44, 46-49, 54-60 and 65-72; the intron elements include a nucleotide sequence selected from the group consisting of SEQ ID NOs 29, 30 and 61; at least a portion of the antibody coding sequence comprises the nucleotide sequence set forth in SEQ ID NO. 62; and/or the polyadenylation sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 31, 33, 34 and 35.

In certain embodiments, the TRE comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 25-27, 36, 39, 42, 44, 46-49, 54-60 and 65-72; the intron elements include a nucleotide sequence selected from the group consisting of SEQ ID NOs 29, 30 and 61; at least a portion of the antibody coding sequence comprises the nucleotide sequence set forth in SEQ ID NO. 63; and/or the polyadenylation sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 31, 33, 34 and 35.

In certain embodiments, the TRE comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 25-27, 36, 39, 42, 44, 46-49, 54-60 and 65-72; the intron elements include a nucleotide sequence selected from the group consisting of SEQ ID NOs 29, 30 and 61; at least a portion of the antibody coding sequence comprises the nucleotide sequence set forth in SEQ ID NO. 83; and/or the polyadenylation sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 31, 33, 34 and 35.

In certain embodiments, the TRE comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 25, 26 or 27; the intron element comprises or consists of the nucleotide sequence shown in SEQ ID NO. 29; at least a portion of the antibody coding sequence comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 52; and/or the polyadenylation sequence comprises or consists of the nucleotide sequence shown in SEQ ID NO. 33.

In certain embodiments, the TRE comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 25, 26 or 27; the intron element comprises or consists of the nucleotide sequence shown in SEQ ID NO. 29; at least a portion of the antibody coding sequence comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 62; and/or the polyadenylation sequence comprises or consists of the nucleotide sequence shown in SEQ ID NO. 33.

In certain embodiments, the TRE comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 25, 26 or 27; the intron element comprises or consists of the nucleotide sequence shown in SEQ ID NO. 29; at least a portion of the antibody coding sequence comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 83; and/or the polyadenylation sequence comprises or consists of the nucleotide sequence shown in SEQ ID NO. 33.

In certain embodiments, the TRE comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 66, 67 or 71; the intron element comprises or consists of the nucleotide sequence shown in SEQ ID NO. 30 or 61; at least a portion of the antibody coding sequence comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 53; and/or the polyadenylation sequence comprises or consists of the nucleotide sequence shown in SEQ ID NO. 31.

In certain embodiments, the TRE comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 66, 67 or 71; the intron element comprises or consists of the nucleotide sequence shown in SEQ ID NO. 30 or 61; at least a portion of the antibody coding sequence comprises or consists of the nucleotide sequence set forth in SEQ ID NO. 63; and/or the polyadenylation sequence comprises or consists of the nucleotide sequence shown in SEQ ID NO. 31.

In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identical to SEQ ID NO 88, 89, 90, or 91. In certain embodiments, the rAAV genome comprises a nucleotide sequence depicted in SEQ ID NO. 88, 89, 90, or 91. In certain embodiments, the rAAV genome consists of the nucleotide sequence depicted in SEQ ID NO 88, 89, 90 or 91.

In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identical to SEQ ID No. 88. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 88. In certain embodiments, the rAAV genome consists of the nucleotide sequence depicted in SEQ ID NO. 88.

In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identical to SEQ ID No. 89. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 89. In certain embodiments, the rAAV genome consists of the nucleotide sequence depicted in SEQ ID NO. 89.

In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identical to SEQ ID NO: 90. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 90. In certain embodiments, the rAAV genome consists of the nucleotide sequence depicted in SEQ ID NO. 90.

In certain embodiments, the rAAV genome comprises a nucleotide sequence that is at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identical to SEQ ID NO. 91. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 91. In certain embodiments, the rAAV genome consists of the nucleotide sequence depicted in SEQ ID NO. 91.

In certain embodiments, the rAAV genomes disclosed herein further comprise a 5 'inverted terminal repeat (5' itr) nucleotide sequence at 5 'relative to the TRE, and a 3' inverted terminal repeat (3 'itr) nucleotide sequence at 3' relative to a polyadenylation sequence associated with the antibody light chain coding sequence. ITR sequences from any AAV serotype or variant thereof can be used in the rAAV genomes disclosed herein. The 5 'and 3' ITRs may be from AAVs of the same serotype or from AAVs of different serotypes. Exemplary ITRs for use in the rAAV genomes disclosed herein are shown in SEQ ID NOs 14, 18, 19, 20, 21 and 32 herein.

In certain embodiments, the 5'itr or 3' itr is from AAV2. In certain embodiments, both the 5'itr and the 3' itr are from AAV2. In certain embodiments, the 5'ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:14, or the 3' ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 18. In certain embodiments, the 5'ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:14, and the 3' ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 18. In certain embodiments, the rAAV genome comprises the nucleotide sequence depicted in SEQ ID NO. 43, the 5'ITR nucleotide sequence has the sequence of SEQ ID NO. 14, and the 3' ITR nucleotide sequence has the sequence of SEQ ID NO. 18.

In certain embodiments, the 5'itr or 3' itr is from AAV5. In certain embodiments, both the 5'itr and the 3' itr are from AAV5. In certain embodiments, the 5'ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:20, or the 3' ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 21. In certain embodiments, the 5'ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:20, and the 3' ITR nucleotide sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 21. In certain embodiments, the rAAV genome comprises a nucleotide sequence depicted in any one of SEQ ID NO. 43, a 5'ITR nucleotide sequence having the sequence of SEQ ID NO. 20, and a 3' ITR nucleotide sequence having the sequence of SEQ ID NO. 21.

In certain embodiments, the 5'itr nucleotide sequence and the 3' itr nucleotide sequence are substantially complementary to each other (e.g., complementary to each other except for mismatches at 1, 2, 3, 4, or 5 nucleotide positions in the 5 'or 3' itr).

In certain embodiments, the 5'ITR or 3' ITR is modified to reduce or eliminate degradation by Rep proteins ("non-cleavable ITR"). In certain embodiments, the non-cleavable ITR comprises an insertion, deletion, or substitution in the nucleotide sequence of the terminal cleavage site. Such modifications allow the formation of a self-complementary double-stranded DNA genome of AAV after replication of the rAAV genome in an infected cell. Exemplary non-cleavable ITR sequences are known in the art (see, e.g., the sequences provided in U.S. patent nos. 7,790,154 and 9,783,824, which are incorporated herein by reference in their entirety). In certain embodiments, the 5' ITR comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO. 19. In certain embodiments, the 5' ITR consists of a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO. 19. In certain embodiments, the 5' ITR consists of the nucleotide sequence set forth in SEQ ID NO. 19. In certain embodiments, the 3' ITR comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO. 32. In certain embodiments, the 5' ITR consists of a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO. 32. In certain embodiments, the 3' ITR consists of the nucleotide sequence set forth in SEQ ID NO. 32. In certain embodiments, the 5'ITR consists of the nucleotide sequence set forth in SEQ ID NO. 19 and the 3' ITR consists of the nucleotide sequence set forth in SEQ ID NO. 32. In certain embodiments, the 5'ITR consists of the nucleotide sequence set forth in SEQ ID NO. 19 and the 3' ITR consists of the nucleotide sequence set forth in SEQ ID NO. 32.

In certain embodiments, the 5' itr flanks an additional nucleotide sequence derived from a wild-type AAV2 genomic sequence. In certain embodiments, the 5' ITR flanks an additional 46bp sequence in the AAV2 genome that is adjacent to the wild-type AAV2 ITR that is derived from the wild-type AAV2 sequence. In certain embodiments, the additional 46bp sequence is 3 'relative to the 5' itr in the rAAV genome. In certain embodiments, the 46bp sequence consists of the nucleotide sequence set forth in SEQ ID NO. 74.

In certain embodiments, the 3' itr flanks an additional nucleotide sequence derived from a wild-type AAV2 genomic sequence. In certain embodiments, the 3' ITR flanks an additional 37bp sequence in the AAV2 genome that is adjacent to the wild-type AAV2 ITR that is derived from the wild-type AAV2 sequence. See, e.g., savy et al, methods of human Gene therapy (Human Gene Therapy Methods) (2017) 28 (5): 277-289, which is incorporated herein by reference in its entirety. In certain embodiments, the additional 37bp sequence is 5 'relative to the 3' itr in the rAAV genome. In certain embodiments, the 37bp sequence consists of the nucleotide sequence set forth in SEQ ID NO. 73.

In another aspect, provided herein are polynucleotides comprising a nucleic acid sequence that is at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to a nucleic acid sequence set forth in SEQ ID No. 84, 85, 86, or 87. In certain embodiments, the polynucleotide comprises or consists of the nucleic acid sequence set forth in SEQ ID NO. 84, 85, 86 or 87.

In another aspect, provided herein are novel rAAV compositions comprising an AAV capsid protein, a rAAV genome as disclosed herein (e.g., a rAAV genome comprising transcriptional regulatory elements operably linked to antibody coding sequences (e.g., antibody heavy chain or light chain coding sequences), thereby allowing for extrachromosomal expression of an antibody in a cell transduced with AAV.

Capsid proteins from any AAV capsid known in the art may be used in the rAAV compositions disclosed herein, including but not limited to capsid proteins from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, or AAV9 serotypes. For example, in certain embodiments, a capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of amino acids 203-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17. In certain embodiments, a capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of amino acids 203-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17, wherein: the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G and the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G. In certain embodiments, the amino acid corresponding to amino acid 296 of SEQ ID NO. 16 in the capsid protein is H, the amino acid corresponding to amino acid 464 of SEQ ID NO. 16 in the capsid protein is N, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 681 of SEQ ID NO. 16 in the capsid protein is M. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A and the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R. In certain embodiments, the amino acid corresponding to amino acid 501 of SEQ ID NO. 16 in the capsid protein is I, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 706 of SEQ ID NO. 16 in the capsid protein is C. In certain embodiments, the capsid protein comprises the amino acid sequence of amino acids 203-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

For example, in certain embodiments, a capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of amino acids 138-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17. In certain embodiments, a capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of amino acids 138-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17, wherein: the amino acid in the capsid protein corresponding to amino acid 151 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 160 of SEQ ID NO. 16 is D; the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G and the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G. In certain embodiments, the amino acid corresponding to amino acid 296 of SEQ ID NO. 16 in the capsid protein is H, the amino acid corresponding to amino acid 464 of SEQ ID NO. 16 in the capsid protein is N, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 681 of SEQ ID NO. 16 in the capsid protein is M. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A and the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R. In certain embodiments, the amino acid corresponding to amino acid 501 of SEQ ID NO. 16 in the capsid protein is I, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 706 of SEQ ID NO. 16 in the capsid protein is C. In certain embodiments, the capsid protein comprises the amino acid sequence of amino acids 138-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

For example, in certain embodiments, a capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of amino acids 1-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17. In certain embodiments, a capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of amino acids 1-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17, wherein: the amino acid in the capsid protein corresponding to amino acid 2 of SEQ ID NO. 16 is T; the amino acid in the capsid protein corresponding to amino acid 65 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 68 of SEQ ID NO. 16 is V; the amino acid in the capsid protein corresponding to amino acid 77 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 119 of SEQ ID NO. 16 is L; the amino acid in the capsid protein corresponding to amino acid 151 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 160 of SEQ ID NO. 16 is D; the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 2 of SEQ ID NO. 16 is T and the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 65 of SEQ ID NO. 16 is I and the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is Y. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 77 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 119 of SEQ ID NO. 16 is L and the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G and the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G. In certain embodiments, the amino acid corresponding to amino acid 296 of SEQ ID NO. 16 in the capsid protein is H, the amino acid corresponding to amino acid 464 of SEQ ID NO. 16 in the capsid protein is N, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 681 of SEQ ID NO. 16 in the capsid protein is M. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R and the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R. In certain embodiments, the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A and the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R. In certain embodiments, the amino acid corresponding to amino acid 501 of SEQ ID NO. 16 in the capsid protein is I, the amino acid corresponding to amino acid 505 of SEQ ID NO. 16 in the capsid protein is R, and the amino acid corresponding to amino acid 706 of SEQ ID NO. 16 in the capsid protein is C. In certain embodiments, the capsid protein comprises the amino acid sequence of amino acids 1-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the AAV capsid comprises two or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID No. 1, 2, 3, 4, 6, 7, 10, 11, 12, 13, 15, 16 or 17; (b) Amino acid sequence capsid proteins comprising amino acids 138-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 15, 16 or 17; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17. In certain embodiments, the AAV capsid comprises: (a) A capsid protein having an amino acid sequence consisting of amino acids 203-736 of SEQ ID No. 1, 2, 3, 4, 6, 7, 10, 11, 12, 13, 15, 16 or 17; (b) A capsid protein having an amino acid sequence consisting of amino acids 138-736 of SEQ ID No. 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 15, 16 or 17; and (c) a capsid protein having an amino acid sequence consisting of amino acids 1-736 of SEQ ID NO. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16 or 17.

In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 203-736 of SEQ ID No. 8; (b) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 138-736 of SEQ ID No. 8; and (c) a capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 1-736 of SEQ ID NO. 8. In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 8; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 8; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 8. In certain embodiments, the AAV capsid comprises two or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 8; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 8; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 8. In certain embodiments, the AAV capsid comprises: (a) A capsid protein having an amino acid sequence consisting of amino acids 203-736 of SEQ ID NO. 8; (b) A capsid protein having an amino acid sequence consisting of amino acids 138-736 of SEQ ID NO. 8; and (c) a capsid protein having an amino acid sequence consisting of amino acids 1-736 of SEQ ID NO. 8.

In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 203-736 of SEQ ID No. 11; (b) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 138-736 of SEQ ID No. 11; and (c) a capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 1-736 of SEQ ID NO. 11. In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 11; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 11; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 11. In certain embodiments, the AAV capsid comprises two or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 11; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 11; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 11. In certain embodiments, the AAV capsid comprises: (a) A capsid protein having an amino acid sequence consisting of amino acids 203-736 of SEQ ID NO. 11; (b) A capsid protein having an amino acid sequence consisting of amino acids 138-736 of SEQ ID NO. 11; and (c) a capsid protein having an amino acid sequence consisting of amino acids 1-736 of SEQ ID NO. 11.

In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 203-736 of SEQ ID No. 13; (b) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 138-736 of SEQ ID No. 13; and (c) a capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of amino acids 1-736 of SEQ ID NO. 13. In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 13; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 13; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 13. In certain embodiments, the AAV capsid comprises two or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 13; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 13; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 13. In certain embodiments, the AAV capsid comprises: (a) A capsid protein having an amino acid sequence consisting of amino acids 203-736 of SEQ ID NO. 13; (b) A capsid protein having an amino acid sequence consisting of amino acids 138-736 of SEQ ID NO. 13; and (c) a capsid protein having an amino acid sequence consisting of amino acids 1-736 of SEQ ID NO. 13.

In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence of amino acids 203-736 of SEQ ID No. 16; (b) A capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence of amino acids 138-736 of SEQ ID No. 16; and (c) a capsid protein comprising an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence of amino acids 1-736 of SEQ ID NO. 16. In certain embodiments, the AAV capsid comprises one or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 16; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 16; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 16. In certain embodiments, the AAV capsid comprises two or more of the following: (a) A capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 16; (b) A capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 16; and (c) a capsid protein comprising the amino acid sequence of amino acids 1-736 of SEQ ID NO. 16. In certain embodiments, the AAV capsid comprises: (a) A capsid protein having an amino acid sequence consisting of amino acids 203-736 of SEQ ID NO. 16; (b) A capsid protein having an amino acid sequence consisting of amino acids 138-736 of SEQ ID NO. 16; and (c) a capsid protein having an amino acid sequence consisting of amino acids 1-736 of SEQ ID NO. 16.

The rAAV genomes of the present disclosure can be used to express any antibody heavy chain and antibody light chain known in the art. In one aspect, provided herein is a rAAV genome comprising a TRE operably linked to at least a portion of an antibody coding sequence (e.g., an antibody heavy chain coding sequence and/or an antibody light chain coding sequence). Non-limiting examples of antibodies include anti-C5 antibodies (e.g., elkuizumab, lei Fuli bead monoclonal antibody, and pamez Li Shan antibody (pozelimab)), anti-factor D antibodies (e.g., lanpalizumab (lumkuumab)), anti-mannose binding protein-related serine protease 2 (MASP-2) antibodies (e.g., nano cable Li Shan anti (narcoplimab)), anti-pancreatic schumab antibodies (e.g., ranavimab (lanadelomab)), anti-interleukin 1 beta antibodies (e.g., kanadumab), anti-interferon gamma antibodies (e.g., emalumab)), anti-PCSK 9 antibodies (e.g., eizokuumab (evaluumab) and alikuumab)), anti-factor IX and factor X antibodies (e.g., bispecific antibodies eimerizumab (icizumab)), and anti-VEGF antibodies (e.g., rannizumab).

In another aspect, the present disclosure provides a pharmaceutical composition comprising an AAV as disclosed herein and a pharmaceutically acceptable excipient, adjuvant, diluent, vehicle, or carrier, or combination thereof. A "pharmaceutically acceptable carrier" comprises any material that, when combined with the active ingredients of a composition, retains the biological activity of the ingredients and does not elicit a destructive physiological response (e.g., an undesired immune response). The pharmaceutically acceptable carrier comprises water, phosphate buffered saline, an emulsion (e.g., an oil/water emulsion), and a humectant. Compositions comprising such carriers are formulated by well known conventional methods, such as those shown below: the pharmaceutical science of Remington's Pharmaceutical Sciences, current edition, mark Publishing company of oiston, pennsylvania (Mack Publishing co., easton Pa.) 18042, usa; gennaro (2000) ", leimington: pharmaceutical science and practice (Remington: the Science and Practice of Pharmacy), "20 th edition, lipping kott. Willust. Wilkins press (lipkincott, williams, & Wilkins); pharmaceutical dosage forms and drug delivery systems (Pharmaceutical Dosage Forms and Drug Delivery Systems) (1999) H.C.Ansel et al, 7 th edition, liPing Cort Williams Wills Press; handbook of pharmaceutical excipients (2000) A.H.Kibbe et al, 3 rd edition, american society of pharmacies.

III methods of use

In another aspect, the disclosure provides methods for expressing antibodies (e.g., antibody heavy and light chains) in cells. The methods generally comprise transducing a cell with a rAAV as disclosed herein. Such methods result in high levels of expression and secretion of antibodies. Thus, in certain embodiments, the methods disclosed herein involve transducing a cell with a rAAV as disclosed herein.

The methods disclosed herein can be applied to any cell (e.g., hepatocyte) where expression of an antibody is desired. Thus, in certain embodiments, the methods are applied to cells in the liver. In certain embodiments, the method is applied to hepatocytes.

The methods disclosed herein may be performed in vitro for research purposes or ex vivo or in vivo for therapeutic purposes.

In certain embodiments, the cells to be transduced are in a mammalian subject and AAV is administered to the subject in an amount effective to transduce the cells in the subject. Thus, in certain embodiments, the present disclosure provides a method of treating a subject suffering from a disease or disorder that would benefit from expression and secretion of an antibody that specifically binds to a therapeutic target, the method generally comprising administering to the subject an effective amount of a rAAV as disclosed herein. In certain embodiments, the antibody specifically binds complement C5 and the disease or disorder is associated with complement C5 activity. The subject may be a human subject or a rodent subject (e.g., a mouse) containing human hepatocytes. Suitable mouse subjects include, but are not limited to, mice into which human hepatocytes (e.g., human hepatocytes) have been implanted. Any disease or disorder associated with complement C5 activity can be treated using the methods disclosed herein. Suitable diseases or conditions include, but are not limited to, paroxysmal sleep hemoglobinuria (PNH), neuromyelitis optica (NMOSD), atypical hemolytic uremic syndrome (aHUS), myasthenia gravis, hematopoietic stem cell transplantation-graft related thrombotic microangiopathy (HSCT-TMA), complement-mediated thrombotic microangiopathy (CM-TMA), guillain-barre syndrome (Guillain-barre syndrome), amyotrophic Lateral Sclerosis (ALS), primary Progressive Multiple Sclerosis (PPMS), multifocal motor neuropathy, antibody-mediated renal rejection, C3 glomerulopathy, age-related macular degeneration (AMD), AQP4 IgG positive neuromyelitis optica, systemic lupus erythematosus, psoriasis, rheumatoid Arthritis (RA), dermatomyositis, idiopathic membranous glomerulopathy, demyelinating neuropathy, complement hyperactivation, vasculogenic thrombosis, protein loss bowel disease (cha) syndrome, geographic Atrophy (GA), asthma, hyperplasia nephritis, and sepsis.

In certain embodiments, the disclosure provides methods for treating a subject having a disease or disorder associated with complement C5 activity, the methods generally comprising administering to the subject an effective amount of a rAAV as disclosed herein. The subject may be a human subject, a non-human primate subject (e.g., cynomolgus monkey), or a rodent subject (e.g., mouse) with aberrant complement C5 activity. Any disease or disorder associated with complement C5 activity can be treated using the methods disclosed herein. Suitable diseases or conditions include, but are not limited to PNH, NMOSD, aHUS, myasthenia gravis, HSCT-TMA, CM-TMA, guillain-Barre syndrome, ALS, PPMS, multifocal motor neuropathy, antibody-mediated renal rejection, C3 glomerulopathy, AMD, AQP4 IgG positive neuromyelitis optica, systemic lupus erythematosus, psoriasis, RA, dermatomyositis, idiopathic membranous glomerulopathy, demyelinating neuropathy, CHAPLE syndrome, geographic Atrophy (GA), asthma, proliferative nephritis, and sepsis.

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 16, and a rAAV genome comprising the following genetic elements from 5 'to 3': a 5'ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 60), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), a T2A peptide cleavage sequence (e.g., a T2A peptide cleavage sequence set forth in SEQ ID NO: 28), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., an SV40 polyadenylation sequence set forth in SEQ ID NO: 31), and a 3'ITR (e.g., a 3' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 18).

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 16, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: a 5'ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 27), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO: 29), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), a T2A peptide cleavage sequence (e.g., a T2A peptide cleavage sequence of SEQ ID NO: 28), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., SV40 polyadenylation sequence of SEQ ID NO: 31), and a 3'ITR (e.g., a 3' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 18).

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of SEQ ID NO. 16, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: a 5' ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 27), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO: 29), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), a polyadenylation sequence (e.g., a BGH polyadenylation sequence of SEQ ID NO: 33), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 67), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO:30 or 61), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., an SV40 polyadenylation sequence of SEQ ID NO: 31), and a nucleotide sequence set forth in SEQ ID NO: 3' ITR (e.g., SEQ ID NO: 18).

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of SEQ ID NO. 16, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: 5'ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a polyadenylation sequence (e.g., a BGH polyadenylation sequence set forth in SEQ ID NO: 33), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO: 29), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 27), a stuffer sequence (e.g., a stuffer comprising the nucleotide sequence set forth in SEQ ID NO: 51), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 67), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO:30 or 61), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 31), and an SV 3 'nucleotide sequence set forth in SEQ ID NO:18, e.g., an SV 3' nucleotide sequence set forth in the SEQ ID NO: 13.

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of amino acids 203-736 of SEQ ID NO. 13, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: a 5'ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 60), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), a T2A peptide cleavage sequence (e.g., a T2A peptide cleavage sequence set forth in SEQ ID NO: 28), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., an SV40 polyadenylation sequence set forth in SEQ ID NO: 31), and a 3'ITR (e.g., a 3' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 18).

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of amino acids 138-736 of SEQ ID NO. 13, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: a 5'ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 27), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO: 29), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), a T2A peptide cleavage sequence (e.g., a T2A peptide cleavage sequence of SEQ ID NO: 28), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., SV40 polyadenylation sequence of SEQ ID NO: 31), and a 3'ITR (e.g., a 3' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 18).

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of SEQ ID NO. 13, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: a 5' ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 27), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO: 29), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), a polyadenylation sequence (e.g., a BGH polyadenylation sequence of SEQ ID NO: 33), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 67), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO:30 or 61), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., an SV40 polyadenylation sequence of SEQ ID NO: 31), and a nucleotide sequence set forth in SEQ ID NO: 3' ITR (e.g., SEQ ID NO: 18).

In certain embodiments, the foregoing methods employ a rAAV comprising an AAV capsid protein comprising the amino acid sequence of SEQ ID NO. 13, and a rAAV genome comprising, 5 'to 3' of the following genetic elements: 5'ITR (e.g., a 5' ITR comprising the nucleotide sequence set forth in SEQ ID NO: 14), a polyadenylation sequence (e.g., a BGH polyadenylation sequence set forth in SEQ ID NO: 33), at least a portion of an antibody heavy chain coding sequence (e.g., an antibody heavy chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:52, 62 or 83), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO: 29), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 27), a stuffer sequence (e.g., a stuffer comprising the nucleotide sequence set forth in SEQ ID NO: 51), a transcriptional regulatory element (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 67), an intron element (e.g., an intron element comprising the nucleotide sequence set forth in SEQ ID NO:30 or 61), at least a portion of an antibody light chain coding sequence (e.g., an antibody light chain coding sequence comprising the nucleotide sequence set forth in SEQ ID NO:53 or 63), a polyadenylation sequence (e.g., a TRE comprising the nucleotide sequence set forth in SEQ ID NO: 31), and an SV 3 'nucleotide sequence set forth in SEQ ID NO:18, e.g., an SV 3' nucleotide sequence set forth in the SEQ ID NO: 13.

The methods disclosed herein are particularly advantageous because they are capable of efficiently expressing antibodies in vivo and secreting into the serum of a subject. In certain embodiments, the serum concentration of the antibody is at least about 100 μg/mL, at least about 500 μg/mL, at least about 1000 μg/mL, at least about 1500 μg/mL, at least about 2000 μg/mL, at least about 2500 μg/mL, at least about 3000 μg/mL, at least about 3500 μg/mL, at least about 4000 μg/mL, at least about 4500 μg/mL, at least about 5000 μg/mL, at least about 7500 μg/mL, at least about 10000 μg/mL, at least about 15000 μg/mL, at least about 20000 μg/mL, at least about 25000 μg/mL, at least about 30000 μg/mL, at least about 35000 μg/mL, at least about 40000 μg/mL, at least about 45000 μg/mL, at least about 50000 μg/mL, at least about 60000 μg/mL, at least about 70000 μg/mL, at least about 80000 μg/mL, at least about 90000 μg/mL, or at least about 100000 μg/mL. Any method of determining the expression level or serum concentration of an antibody may be used, including but not limited to ELISA, western blot, immunostaining, and mass spectrometry. In certain embodiments, the serum concentration of the antibody is determined using an anti-human IgG ELISA.

In certain embodiments, transducing cells with an AAV composition disclosed herein can be performed as provided herein or by any transduction method known to one of skill in the art. In certain embodiments, the cells may be at 50,000;100,000;150,000;200,000;250,000;300,000;350,000;400,000;450,000; or a multiplicity of infection (MOI) of 500,000 or any MOI that provides optimal transduction of cells is contacted with AAV.

AAV compositions disclosed herein may be administered to a subject by any suitable route, including but not limited to intravenous, intraperitoneal, subcutaneous, intramuscular, intranasal, topical, or intradermal routes. In certain embodiments, the compositions are formulated for administration by intravenous injection or subcutaneous injection.

AAV packaging system

In another aspect, the present disclosure provides packaging systems for recombinantly producing a recombinant adeno-associated virus (rAAV) as disclosed herein. Such packaging systems generally comprise: a first nucleotide encoding one or more AAV Rep proteins; a second nucleotide encoding a capsid protein of any AAV of the AAVs as disclosed herein; and a third nucleotide sequence comprising any of the rAAV genomes as disclosed herein, wherein the packaging system is operable in a cell to enclose the rAAV genome in a capsid to form an AAV.

In certain embodiments, the packaging system comprises a first vector comprising a first nucleotide sequence encoding one or more AAV Rep proteins and a second nucleotide sequence encoding an AAV capsid protein, and a second vector comprising a third nucleotide sequence comprising a rAAV genome. As used in the context of packaging systems as described herein, "vector" refers to a nucleic acid molecule (e.g., plasmid, virus, cosmid, artificial chromosome, etc.) that is a vehicle for introducing nucleic acid into cells.

Any AAV Rep protein may be used in the packaging systems disclosed herein. In certain embodiments of the packaging system, the Rep nucleotide sequence encodes an AAV2 Rep protein. Suitable AAV2 Rep proteins include, but are not limited to, rep 78/68 or Rep 68/52. In certain embodiments of the packaging system, the nucleotide sequence encoding an AAV2 Rep protein comprises a nucleotide sequence encoding a protein having a minimal percent sequence identity to the AAV2 Rep amino acid sequence of SEQ ID No. 22, wherein the minimal percent sequence identity across the length of the amino acid sequence of the AAV2 Rep protein is at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%). In certain embodiments of the packaging system, the AAV2 Rep protein has the amino acid sequence depicted in SEQ ID NO. 22.

In certain embodiments of the packaging system, the packaging system further comprises a fourth nucleotide sequence comprising one or more helper virus genes. In certain embodiments of the packaging system, the packaging system further comprises a third vector, e.g., a helper vector, comprising a fourth nucleotide sequence comprising one or more helper genes. The third vector may be a separate third vector, integrated with the first vector, or integrated with the second vector.

In certain embodiments of the packaging system, the helper virus is selected from the group consisting of: adenoviruses, herpesviruses (including Herpes Simplex Virus (HSV)), poxviruses (e.g., vaccinia virus), cytomegalovirus (CMV), and baculoviruses. In certain packaging system embodiments wherein the helper virus is an adenovirus, the adenovirus genome comprises one or more adenovirus RNA genes selected from the group consisting of: el, E2, E4 and VA. In certain packaging system embodiments in which the helper virus is HSV, the HSV genome comprises one or more of the HSV genes selected from the group consisting of: UL5/8/52, ICPO, ICP4, ICP22 and UL30/UL42.

In certain embodiments of the packaging system, the first vector, the second vector, and/or the third vector are contained within one or more plasmids. In certain embodiments, the first vector and the third vector are contained within a first plasmid. In certain embodiments, the second vector and the third vector are contained within a second plasmid.

In certain embodiments of the packaging system, the first, second, and/or third vectors are contained within one or more recombinant helper viruses. In certain embodiments, the first vector and the third vector are contained within a recombinant helper virus. In certain embodiments, the second vector and the third vector are contained within a recombinant helper virus.

In another aspect, the disclosure provides a method for recombinant production of AAV as described herein, wherein the method comprises transfecting or transducing a cell with a packaging system as described herein under conditions operable to encapsulate a rAAV genome in a capsid to form a rAAV as described herein. Exemplary methods for recombinant production of a rAAV comprise transient transfection (e.g., with one or more transfection plasmids containing a first vector and a second vector and optionally a third vector as described herein), viral infection (e.g., with one or more recombinant helper viruses, such as adenovirus, poxvirus (e.g., vaccinia virus), herpes virus (including HSV, cytomegalovirus, or baculovirus, containing a first vector and a second vector and optionally a third vector as described herein), and transfection or infection of a stable producer cell line (e.g., with a stable producer cell, such as a mammalian or insect cell, containing a Rep nucleotide sequence encoding one or more AAV Rep proteins and/or a Cap nucleotide sequence encoding one or more AAV capsid proteins as described herein), and wherein the rAAV genome as described herein is delivered in plasmid or recombinant helper virus form.

Accordingly, the present disclosure provides a packaging system for preparing a recombinant AAV (rAAV), wherein the packaging system comprises a first nucleotide sequence encoding one or more AAV Rep proteins; a second nucleotide sequence encoding a capsid protein of any one of the AAV described herein; a third nucleotide sequence comprising a rAAV genomic sequence of any one of the AAV described herein; and optionally a fourth nucleotide sequence comprising one or more helper virus genes.

V. examples

The following examples demonstrate the efficient expression of antibodies (e.g., anti-C5 antibodies) in a subject using rAAV vectors as disclosed herein. These examples are provided by way of illustration and not by way of limitation.

Example 1: anti-complement C5 antibody rAAV vector

The present example provides anti-C5 antibody expression vectors C5Ab01, C5Ab02, C5Ab03, and C5Ab04 for expressing an anti-C5 antibody in cells transduced with the vectors (e.g., human cells or mouse cells).

a)C5Ab01

As shown in fig. 1, the anti-C5 antibody vector C5Ab01 comprises the following genetic elements from 5 'to 3': a transcriptional regulatory element comprising an EF1 a promoter; a coding sequence encoding a human IgG2 (P1) signal sequence linked to an anti-C5 antibody Heavy Chain (HC); a nucleic acid sequence encoding a 2A ribosome-skipping peptide; a coding sequence encoding an igκ (P2) signal sequence linked to an anti-C5 antibody Light Chain (LC); and SV40 late polyadenylation sequences (LPA). The nucleic acid sequences of these elements are shown in table 1. This vector is capable of expressing an anti-C5 antibody in a cell (e.g., a human cell or a mouse cell) into which the vector is transduced.

b)C5Ab02

As shown in fig. 1, the anti-C5 antibody vector C5Ab02 comprises the following genetic elements from 5 'to 3': transcriptional regulatory elements including a liver-specific LP1 promoter; a coding sequence encoding a human IgG2 (P1) signal sequence linked to an anti-C5 antibody Heavy Chain (HC); a nucleic acid sequence encoding a 2A ribosome-skipping peptide; a coding sequence encoding an igκ (P2) signal sequence linked to an anti-C5 antibody Light Chain (LC); and SV40 late polyadenylation sequences (LPA). The sequences of these elements are shown in table 1. This vector is capable of expressing an anti-C5 antibody in a cell (e.g., a human cell or a mouse cell) into which the vector is transduced.

c)C5Ab03

As shown in fig. 1, the anti-C5 antibody vector C5Ab03 comprises the following genetic elements from 5 'to 3': transcriptional regulatory elements including a liver-specific LP1 promoter; a coding sequence encoding a human IgG2 (P1) signal sequence linked to an anti-C5 antibody Heavy Chain (HC); bovine growth hormone polyadenylation signal (bGHpA); transcriptional regulatory elements including a liver-specific DnG promoter; a coding sequence encoding an igκ (P2) signal sequence linked to an anti-C5 antibody Light Chain (LC); and SV40 late polyadenylation sequences (LPA). The sequences of these elements are shown in table 1. This vector is capable of expressing an anti-C5 antibody in a cell (e.g., a human cell or a mouse cell) into which the vector is transduced.

d)C5Ab04

As shown in fig. 1, the anti-C5 antibody vector C5Ab04 comprises the following genetic elements from 5 'to 3': bovine growth hormone polyadenylation signal (bGHpA); anti-C5 antibody heavy chain coding sequences; a coding sequence encoding an anti-C5 antibody Heavy Chain (HC) linked to a human IgG2 (P1) signal sequence; transcriptional regulatory elements including a liver-specific LP1 promoter; a filling sequence; transcriptional regulatory elements including a liver-specific DnG promoter; a coding sequence encoding an igκ (P2) signal sequence linked to an anti-C5 antibody Light Chain (LC); and SV40 late polyadenylation sequences (LPA). The sequences of these elements are shown in table 1. This vector is capable of expressing an anti-C5 antibody in a cell (e.g., a human cell or a mouse cell) into which the vector is transduced.

Table 1: genetic elements in anti-C5 antibody expression vectors C5Ab01, C5Ab02, C5Ab03 and C5Ab04

Vectors disclosed herein can be packaged in AAV capsids, including but not limited to AAVHSC5, AAVHSC7, AAVHSC8, AAVHSC13, AAVHSC15, or AAVHSC17 capsids.

Example 2: expression of anti-C5 antibodies in mouse models

Abnormal or excessive activity of complement component C5 is associated with several diseases, including paroxysmal sleep hemoglobinuria (PNH), neuromyelitis optica spectrum disorders (NMOSD), and atypical hemolytic uremic syndrome (aHUS). anti-C5 monoclonal antibodies have been shown to be effective in treating these diseases, but patients often require multiple large doses of antibodies to obtain therapeutic benefit. This is due in part to the high concentration of C5 protein in the patient's serum. Thus, high levels of anti-C5 antibody binding and elimination of sufficient C5 are required to produce the desired therapeutic effect. These problems can be overcome if the patient is able to express his own anti-C5 antibody.

To investigate whether a sufficiently high level of anti-C5 antibody could be expressed in organisms, AAV vectors for expressing the anti-C5 antibody from mouse liver were administered to NOD SCID mice. Four separate experiments were performed, test vectors C5Ab02, C5Ab03, and C5Ab04 (described above) packaged in each of AAVHSC13, AAVHSC15, and AAVHSC 17.

Human IgG ELISA protocol

To assess serum human IgG concentration (μg/mL) in the following experiments, simpleStep from Abcam was usedA kit. Briefly, an antibody mixture is prepared by diluting the capture and detection antibodies in an antibody diluent CP. To prepare 3mL of an antibody mixture, 300. Mu.L of 10X capture antibody and 300. Mu.L of 10X detection antibody were combined with2.4mL antibody diluent CP combination. Standards were then prepared at the point of use by serial dilution. The human IgG protein provided in the kit was used for positive control serial dilutions.

For analysis, all reagents were brought to room temperature prior to use. All samples or standards of 50 μl were added to the appropriate wells of the microwell plate. Then 50 μl of the antibody mixture was added to each well. The plates were then sealed and incubated at room temperature for 40 minutes on a plate shaker set at 400 rpm. Each well was then washed with 3X 350. Mu.L of 1 Xwashing buffer PT. After the last wash, the plate was inverted and blotted with a clean paper towel to remove excess liquid. Then 100. Mu.L of TMB developing solution was added to each well and incubated in the dark for about 5 minutes on a plate shaker set at 400 rpm. The optimal incubation time may vary between 5 minutes and 20 minutes. To each well 100 μl of stop solution was added. The plates were shaken on a plate shaker for 1 minute to mix. The Optical Density (OD) was then read at 450 nm. This is the endpoint reading.

Results

In the first experiment, mice received a 1e13 vg/kg dose of vector C5Ab04 packaged in AAVHSC13 or AAVHSC17 capsids. Serum samples were collected after 1 week, 3 weeks, 5 weeks, 7 weeks, 9 weeks, 11 weeks, 15 weeks, 19 weeks and 23 weeks. Serum samples were tested for human IgG concentration (μg/mL) as a reading of anti-C5 antibody levels. Female mice are poor models of AAV-mediated gene transfer, thus segregating data between male and female mice. As shown in fig. 2A, mice that received C5Ab04 packaged in AAVHSC13 or AAVHSC17 capsids showed elevated levels of anti-C5 antibodies over time. Fig. 2B shows the results in fig. 2A, with the Y-axis on a logarithmic scale. In fig. 2A-2B, n=2-3 mice/group.

In a second experiment, mice received a dose of 1e13 vgs/kg of vector C5Ab02 packaged in AAVHSC17 capsid. Data from male and female mice were isolated and multiple serum samples were collected over a 16 week period. Serum samples were tested for human IgG concentration (μg/mL) as a reading of anti-C5 antibody levels. As shown in fig. 2C, mice that received vector C5Ab02 packaged in AAVHSC17 capsids showed elevated levels of anti-C5 antibodies over time. Fig. 2D shows the results in fig. 2C, with the Y-axis on a logarithmic scale. In fig. 2C-2D, n=3 mice/group.

In a third experiment, mice received 1e13 vg/kg dose of vector C5Ab02, C5Ab03, or C5Ab04 packaged in AAVHSC15 or AAVHSC17 capsids, respectively. Data from male mice are shown and multiple serum samples are collected over a 16 week period. Serum samples were tested for human IgG concentration (μg/mL) as a reading of anti-C5 antibody levels. As shown in fig. 2E, mice that received any of the vectors C5Ab02, C5Ab03, or C5Ab04 packaged in AAVHSC15 or AAVHSC17 capsids showed elevated levels of anti-C5 antibodies over time. Fig. 2F and 2G (logarithmic scale on Y-axis) show the results in fig. 2E, presented in line graph format. In figure 2E-2g, n=3 male mice/group.

In a fourth experiment, mice received 5 doses of 5e11 vg/kg, 5e12 vg/kg, 1.4e13 vg/kg, 4.4e13 vg/kg, and 1.8e14 vg/kg of vector C5Ab04 packaged in AAVHSC17 capsid. Data from male mice are shown and multiple serum samples are collected over a 13 week period. Serum samples were tested for human IgG concentration (μg/mL) as a reading of anti-C5 antibody levels. As shown in the dose-response data of fig. 2H, mice that received vector C5Ab04 packaged in AAVHSC17 capsids showed elevated anti-C5 antibody levels over time. Furthermore, increasing the dosage of AAVHSC17 resulted in a corresponding increase in the concentration of anti-C5 antibody, with dosages of 1.4e13 vg/kg, 4.4e13 vg/kg, and 1.8e14 vg/kg achieving the mg concentration of antibody. Fig. 2I shows the results in fig. 2H presented in line graph format with the Y-axis on a logarithmic scale. In fig. 2E-2I, n=3 male mice/group.

The above data were reorganized to compare the efficacy of each vector packaged in each AAVHSC13, AAVHSC15 or AAVHSC 17. As shown in fig. 3A-3C, C5Ab04 consistently produced higher concentrations of anti-C5 antibodies over time. Furthermore, AAVHSC17 capsids consistently produced higher concentrations of anti-C5 antibodies over time. The combination of C5Ab04 packaged in AAVHSC17 capsid produced the highest concentration of anti-C5 antibody 15 weeks after delivery to mice, approximately 2000 μg/mL.

The above data are also used to predict whether the concentration of anti-C5 antibody achieved is consistent with known therapeutic anti-C5 antibodies administered directly to the patient (i.e., not expressed from a vector in the patient). Based on C for the treatment of PNH _max 、C _{Cereal grain} And dosing schedule data, pharmacokinetic modeling of commercially available eculizumab and Lei Fuli bead mab. The standard deviation range is based on the coefficient of variation and the number of patients used for these studies (dashed lines in fig. 4A and 4B). This modeling data was compared to the data from the in vivo mouse experiments described above for AAVHSC13 and AAVHSC17 packaging vectors from the first experiment. Comparison shows that the therapeutic methods described herein can achieve clinically effective levels of the known anti-C5 antibodies Lei Fuli bead mab and eculizumab (fig. 4A). The modeling data were also compared with data from the in vivo mouse experiments described above (first experiment: "NOD-SCID,1E+13vg/kg"; and fourth experiment: "NOD-SCID 1.8E+14vg/kg") and with data from the HuLiv mouse experiment (example 4) below for C5Ab04 packaged in AAVHSC17 capsid (FIG. 4B).

Example 3: in vitro analysis of anti-C5 antibodies expressed in mouse models

Paroxysmal sleep hemoglobinuria (PNH) is characterized by destruction of erythrocytes (hemolytic anemia), blood clots (thrombus), and impaired bone marrow function. To determine whether the expressed anti-C5 antibodies were effective in reducing hemolytic anemia, PNH ex vivo hemolysis assays were performed. Human serum containing human C5 was mixed with serum obtained from NOD SCID mice treated with AAVHSC17 packaged C5Ab04 and activated sheep Red Blood Cells (RBCs). The% hemolysis was compared to a control anti-C5 biomimetic antibody. As shown in fig. 5, serum obtained from NOD SCID mice treated with AAVHSC 13-packaged C5Ab04 or AAVHSC 17-packaged C5Ab04 was able to inhibit hemolysis to a greater extent than the biomimetic control anti-C5 antibody.

Similar ex vivo hemolysis assays were performed comparing sera obtained from NOD SCID mice treated with 1e13 vg/kg dose of AAVHSC13 packaged C5Ab04 or AAVHSC17 packaged C5Ab04 with negative control mouse sera (from the experiment in fig. 2A). Data from male and female mice were isolated and multiple serum samples were collected over a 9 week period after treatment. Human IgG concentrations (μg/mL) were measured in serum samples as a read of anti-C5 antibody levels, and increased anti-C5 antibody levels over time were confirmed in mice treated with C5Ab04 packaged in AAVHSC13 or AAVHSC17 capsids (fig. 6A). The data presented in fig. 6A is a subset of the data presented in fig. 2A and 2B. Serum obtained from mice treated with C5Ab04 packaged in AAVHSC13 or AAVHSC17 capsids was able to inhibit haemolysis (fig. 6B). Fig. 6C shows the results in fig. 6B, wherein% hemolysis was determined from serum samples obtained 19 weeks after administration and presented in line graphs. In fig. 6A-6C, n=3 mice/group.

In a second experiment, an ex vivo hemolysis assay was performed comparing serum obtained from NOD SCID mice treated with C5Ab02 packaged with AAVHSC17 at a dose of 1e13 vg/kg with negative control mouse serum (as in fig. 2C). Data from male and female mice were isolated and multiple serum samples were collected over a 16 week period after treatment. As shown in fig. 6D, serum obtained from treated NOD SCID mice was able to inhibit hemolysis. In fig. 6D, n=3 mice/group.

In a third experiment, an ex vivo hemolysis assay was performed comparing serum obtained from male NOD SCID mice treated with 1E13 vg/kg dose of AAVHSC15 or AAVHSC17 packaged C5Ab02, C5Ab03 or C5Ab04 with negative control mouse serum (as in fig. 2E). Multiple serum samples were collected over a period of 16 weeks after treatment. As shown in fig. 6E, serum obtained from treated NOD SCID mice was able to inhibit hemolysis. In fig. 6E, n=3 mice/group.

In a fourth experiment, an ex vivo hemolysis assay was performed comparing serum obtained from male NOD SCID mice treated with C5Ab04 packaged with AAVHSC17 at a dose of 5e11 vg/kg, 5e12 vg/kg, 1.4e13 vg/kg, 4.4e13 vg/kg, or 1.8e14 vg/kg with negative control mouse serum (as in fig. 2H). Multiple serum samples were collected over a period of 16 weeks after treatment. As shown in fig. 6F, serum obtained from treated NOD SCID mice was able to inhibit hemolysis in a rAAV dose-dependent manner. In fig. 6F, n=3 mice/group.

Hemolysis assay protocol

The hemolysis assay mentioned above was performed using the following protocol. Gelatin fluxolone buffer (Gelatin Veronal buffer, GVBS, sigma, catalog No. G6514) was mixed with mouse serum in 96-well V-bottom plates in each well (with and without EDTA). 10% normal human serum (NHS, sigma, catalog number H4522) was then added to all wells. The plates were then incubated at 37℃for 30 minutes. 1mL of antibody-sensitized sheep red blood cells (Complement Technology, cat# B200, B201 and B202) were then added to each well and shaken for about 30 minutes. The plates were then centrifuged at 1000g for 5 minutes. The supernatant was moved to a new plate and read at 540nm and 615 nm. The 615nm value was then subtracted from the 540nm value to obtain the final reading. In all reported% hemolysis values, all samples, including% hemolysis of buffer only formulation or AAVHSC treated samples, were reported after normalization with 100% erythrocyte lysis control.

Example 4: generation and characterization of HuLiv mouse model

To evaluate the functional activity and durability of the therapeutic methods described herein, the general term in the context of C57Bl/6 is usedFah of knockout mice ^-/- Rag2 ^-/- Il2rg ^-/- Mice were used as a liver humanization model. Mice are immunodeficient and lack the tyrosine catabolic enzyme fumarylacetoacetate hydrolase (Fah). Ablation of mouse hepatocytes was induced by extraction of the protective drug 2- (2-nitro-4-trifluoromethylbenzoyl) -1, 3-cyclohexanedione (NTBC). The mice are then transplanted with human hepatocytes and adenovirus expressing urokinase is administered to enhance reconstitution of the human hepatocytes. With an appropriate CuRx ^TM Nitisinone (20-0026) and a prophylactic treatment (20-0037) regimen of SMX/TMP antibiotics maintained transplantation throughout the life of the animals. The obtained HuLiv mouse is used for liver>70% of human hepatocytes are reconstituted. These HuLiv mice are further described in the following: azuma et al (Nature Biotechnology 25 (8): 903-910 (2007)).

As shown in fig. 7A and 7B, huLiv mice produced human C5 levels comparable to human serum, while producing less mouse C5. This HuLiv model allows for examination of human hepatocytes in vivo for the expression of anti-C5 antibodies and the durability of anti-C5 antibodies in the presence of human C5.

HuLiv mice (n=2) were administered a single dose of 100 μg of control anti-C5 biomimetic antibody or 1e13 vg/kg or 1e14 vg/kg of C5Ab04 packaged in AAVHSC17 capsids. Serum antibody concentrations were measured at week 1, week 3, week 5, week 7, week 9 and week 11. As shown in fig. 8A, administration of C5Ab04 packaged in AAVHSC17 capsids to HuLiv mice resulted in substantially higher concentrations of anti-C5 antibodies for weeks 1, 3, and 5 compared to direct administration of control anti-C5 antibodies. Fig. 8B shows the results in fig. 8A, wherein serum antibody concentrations were determined 11 weeks after administration and presented in line plots with the Y-axis on a logarithmic scale. The ex vivo hemolysis assay was performed using the same methods as described above comparing serum obtained from HuLiv mice treated with C5Ab04 packaged in AAVHSC17 capsids at a dose of 1e13 vg/kg or 1e14 vg/kg with serum obtained from HuLiv mice administered with control anti-C5 antibodies (bio-mimetic pharmaceuticals). Serum samples were collected over a period of 11 weeks after treatment. As shown in fig. 8C, serum obtained from treated HuLiv mice was able to inhibit hemolysis. Mice treated with 1e13 vg/kg or 1e14 vg/kg dose of C5Ab04 packaged in AAVHSC17 capsid showed about 80% protection from hemolysis in an ex vivo hemolysis assay. Up to about 20% of background residual hemolysis can be explained by the presence of mouse complement proteins prepared from a residual population of C57Bl/6 hepatocytes in Huliv mice. Fig. 8D shows mouse C5 levels detected in serum obtained from treated HuLiv mice via enzyme-linked immunosorbent assay.

Example 5: primary hepatocyte screening assay

To rapidly test optimized rAAV vectors, cell line-based assays were developed to assess antibody production and secretion. Human primary hepatocytes were selected as the closest match to in vivo experiments. Spreadable human hepatocytes (catalog number HUCPG) and spreadable C57BL/6 mouse hepatocytes (catalog number MBCP 01) from Lonza were used.

Approximately 500,000 human hepatocytes or 250,000 mouse hepatocytes were plated and then incubated with about 300,000MOI AAVHSC15 or AAVHSC17 packaged C5Ab02, C5Ab03, or C5Ab 04. Media was then collected on day 7 after virus addition and analyzed by western blot and human IgG ELISA. As shown in fig. 9A and 9B, a large number of levels of anti-C5 antibodies were detected.

***

The present invention should not be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

All references (e.g., publications or patents or patent applications) cited herein are hereby incorporated by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the following claims.

Sequence listing

<110> homologous pharmaceutical Co (HOMOLOGY MEDICINES, INC.)

<120> vectorized antibodies and uses thereof

<130> 404217-HMW-042WO (185407)

<150> US 63/075,898

<151> 2020-09-09

<150> US 63/179,990

<151> 2021-04-26

<160> 132

<170> PatentIn version 3.5

<210> 1

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 1

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 2

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 2

Met Thr Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Gln Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 3

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 3

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Gly Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Gly Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 4

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 4

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Ile Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Tyr Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 5

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 5

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Asp

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 6

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 6

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Leu Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Ser Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 7

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 7

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Arg Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 8

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 8

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Val Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 9

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 9

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Arg Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 10

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 10

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Cys Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 11

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 11

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Arg Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Lys Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 12

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 12

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro His Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Asn

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Arg Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Met Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 13

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 13

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Arg Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 14

<211> 145

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 14

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcct 145

<210> 15

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 15

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Arg Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Arg Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 16

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 16

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Ala Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Arg Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 17

<211> 736

<212> PRT

<213> artificial sequence

<220>

<223> adeno-associated virus

<400> 17

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Ile Ala Trp Pro Arg Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Cys Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 18

<211> 145

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 18

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag agagggagtg gccaa 145

<210> 19

<211> 106

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 19

ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60

ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtgg 106

<210> 20

<211> 167

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 20

ctctcccccc tgtcgcgttc gctcgctcgc tggctcgttt gggggggtgg cagctcaaag 60

agctgccaga cgacggccct ctggccgtcg cccccccaaa cgagccagcg agcgagcgaa 120

cgcgacaggg gggagagtgc cacactctca agcaaggggg ttttgta 167

<210> 21

<211> 167

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 21

tacaaaacct ccttgcttga gagtgtggca ctctcccccc tgtcgcgttc gctcgctcgc 60

tggctcgttt gggggggtgg cagctcaaag agctgccaga cgacggccct ctggccgtcg 120

cccccccaaa cgagccagcg agcgagcgaa cgcgacaggg gggagag 167

<210> 22

<211> 621

<212> PRT

<213> adeno-associated Virus 2

<400> 22

Met Pro Gly Phe Tyr Glu Ile Val Ile Lys Val Pro Ser Asp Leu Asp

1 5 10 15

Glu His Leu Pro Gly Ile Ser Asp Ser Phe Val Asn Trp Val Ala Glu

20 25 30

Lys Glu Trp Glu Leu Pro Pro Asp Ser Asp Met Asp Leu Asn Leu Ile

35 40 45

Glu Gln Ala Pro Leu Thr Val Ala Glu Lys Leu Gln Arg Asp Phe Leu

50 55 60

Thr Glu Trp Arg Arg Val Ser Lys Ala Pro Glu Ala Leu Phe Phe Val

65 70 75 80

Gln Phe Glu Lys Gly Glu Ser Tyr Phe His Met His Val Leu Val Glu

85 90 95

Thr Thr Gly Val Lys Ser Met Val Leu Gly Arg Phe Leu Ser Gln Ile

100 105 110

Arg Glu Lys Leu Ile Gln Arg Ile Tyr Arg Gly Ile Glu Pro Thr Leu

115 120 125

Pro Asn Trp Phe Ala Val Thr Lys Thr Arg Asn Gly Ala Gly Gly Gly

130 135 140

Asn Lys Val Val Asp Glu Cys Tyr Ile Pro Asn Tyr Leu Leu Pro Lys

145 150 155 160

Thr Gln Pro Glu Leu Gln Trp Ala Trp Thr Asn Met Glu Gln Tyr Leu

165 170 175

Ser Ala Cys Leu Asn Leu Thr Glu Arg Lys Arg Leu Val Ala Gln His

180 185 190

Leu Thr His Val Ser Gln Thr Gln Glu Gln Asn Lys Glu Asn Gln Asn

195 200 205

Pro Asn Ser Asp Ala Pro Val Ile Arg Ser Lys Thr Ser Ala Arg Tyr

210 215 220

Met Glu Leu Val Gly Trp Leu Val Asp Lys Gly Ile Thr Ser Glu Lys

225 230 235 240

Gln Trp Ile Gln Glu Asp Gln Ala Ser Tyr Ile Ser Phe Asn Ala Ala

245 250 255

Ser Asn Ser Arg Ser Gln Ile Lys Ala Ala Leu Asp Asn Ala Gly Lys

260 265 270

Ile Met Ser Leu Thr Lys Thr Ala Pro Asp Tyr Leu Val Gly Gln Gln

275 280 285

Pro Val Glu Asp Ile Ser Ser Asn Arg Ile Tyr Lys Ile Leu Glu Leu

290 295 300

Asn Gly Tyr Asp Pro Gln Tyr Ala Ala Ser Val Phe Leu Gly Trp Ala

305 310 315 320

Thr Lys Lys Phe Gly Lys Arg Asn Thr Ile Trp Leu Phe Gly Pro Ala

325 330 335

Thr Thr Gly Lys Thr Asn Ile Ala Glu Ala Ile Ala His Thr Val Pro

340 345 350

Phe Tyr Gly Cys Val Asn Trp Thr Asn Glu Asn Phe Pro Phe Asn Asp

355 360 365

Cys Val Asp Lys Met Val Ile Trp Trp Glu Glu Gly Lys Met Thr Ala

370 375 380

Lys Val Val Glu Ser Ala Lys Ala Ile Leu Gly Gly Ser Lys Val Arg

385 390 395 400

Val Asp Gln Lys Cys Lys Ser Ser Ala Gln Ile Asp Pro Thr Pro Val

405 410 415

Ile Val Thr Ser Asn Thr Asn Met Cys Ala Val Ile Asp Gly Asn Ser

420 425 430

Thr Thr Phe Glu His Gln Gln Pro Leu Gln Asp Arg Met Phe Lys Phe

435 440 445

Glu Leu Thr Arg Arg Leu Asp His Asp Phe Gly Lys Val Thr Lys Gln

450 455 460

Glu Val Lys Asp Phe Phe Arg Trp Ala Lys Asp His Val Val Glu Val

465 470 475 480

Glu His Glu Phe Tyr Val Lys Lys Gly Gly Ala Lys Lys Arg Pro Ala

485 490 495

Pro Ser Asp Ala Asp Ile Ser Glu Pro Lys Arg Val Arg Glu Ser Val

500 505 510

Ala Gln Pro Ser Thr Ser Asp Ala Glu Ala Ser Ile Asn Tyr Ala Asp

515 520 525

Arg Tyr Gln Asn Lys Cys Ser Arg His Val Gly Met Asn Leu Met Leu

530 535 540

Phe Pro Cys Arg Gln Cys Glu Arg Met Asn Gln Asn Ser Asn Ile Cys

545 550 555 560

Phe Thr His Gly Gln Lys Asp Cys Leu Glu Cys Phe Pro Val Ser Glu

565 570 575

Ser Gln Pro Val Ser Val Val Lys Lys Ala Tyr Gln Lys Leu Cys Tyr

580 585 590

Ile His His Ile Met Gly Lys Val Pro Asp Ala Cys Thr Ala Cys Asp

595 600 605

Leu Val Asn Val Asp Leu Asp Asp Cys Ile Phe Glu Gln

610 615 620

<210> 23

<211> 57

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 23

atgggctggt cctgcatcat cctgttcctg gtggccaccg ccacaggcgt gcacagc 57

<210> 24

<211> 66

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 24

atggacatga gggtccctgc tcagctgctg gggctcctgc tgctctggct cagcggtgcc 60

agatgt 66

<210> 25

<211> 192

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 25

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gg 192

<210> 26

<211> 255

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 26

aatgactcct ttcggtaagt gcagtggaag ctgtacactg cccaggcaaa gcgtccgggc 60

agcgtaggcg ggcgactcag atcccagcca gtggacttag cccctgtttg ctcctccgat 120

aactggggtg accttggtta atattcacca gcagcctccc ccgttgcccc tctggatcca 180

ctgcttaaat acggacgagg acagggccct gtctcctcag cttcaggcac caccactgac 240

ctgggacagt gaatc 255

<210> 27

<211> 448

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 27

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacagggc cctgtctcct cagcttcagg 420

caccaccact gacctgggac agtgaatc 448

<210> 28

<211> 54

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 28

gaaggcagag gcagcctgct gacctgcggc gacgtcgaag agaaccccgg ccct 54

<210> 29

<211> 93

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 29

ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt 60

ttctctcttt tagattccaa cctttggaac tga 93

<210> 30

<211> 92

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 30

aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag 60

cacctgcctg aaatcacttt ttttcaggtt gg 92

<210> 31

<211> 198

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 31

gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga 60

aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc 120

tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag 180

gtgtgggagg ttttttaa 198

<210> 32

<211> 143

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 32

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag agagggagtg gcc 143

<210> 33

<211> 171

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 33

ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 60

tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 120

tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag g 171

<210> 34

<211> 122

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 34

aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 60

aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 120

ta 122

<210> 35

<211> 133

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 35

tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 60

aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg 120

gaggtttttt aaa 133

<210> 36

<211> 1676

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 36

ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 60

catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 120

acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 180

ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 240

aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 300

ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat 360

tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct 420

cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 480

tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg 540

gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc 600

cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg 660

gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc 720

cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 780

cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa 840

gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg 900

tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct 960

gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg 1020

gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt 1080

gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc 1140

cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg 1200

gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg 1260

ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg 1320

gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc 1380

agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc 1440

cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg 1500

gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc 1560

ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga 1620

ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacag 1676

<210> 37

<211> 12

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 37

agaaagagaa ga 12

<210> 38

<211> 12

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 38

agaagaaaga ga 12

<210> 39

<211> 1168

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 39

cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 60

tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg 120

aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 180

gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa 240

gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt 300

gaattacttc cacctggctc cagtacgtga ttcttgatcc cgagctggag ccaggggcgg 360

gccttgcgct ttaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg 420

gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc 480

tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct 540

tgtaaatgcg ggccaggatc tgcacactgg tatttcggtt tttggggccg cgggcggcga 600

cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc 660

gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc 720

gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc 780

ggaaagatgg ccgcttcccg gccctgctcc agggggctca aaatggagga cgcggcgctc 840

gggagagcgg gcgggtgagt cacccacaca aaggaaaggg gcctttccgt cctcagccgt 900

cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctggag 960

cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca 1020

cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga 1080

atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag 1140

tttttttctt ccatttcagg tgtcgtga 1168

<210> 40

<211> 72

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 40

agatctggca gcggagaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 60

cccggcccta gg 72

<210> 41

<211> 95

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 41

cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg 60

ttattgtgct gtctcatcat tttggcaaag aattc 95

<210> 42

<211> 1873

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 42

gatcttcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt 60

ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca 120

tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata gtaatcaatt 180

acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 240

ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 300

cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 360

actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc 420

aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct 480

acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg 540

ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt 600

ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg 660

gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag 720

agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa 780

aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc 840

cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag 900

cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt 960

ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg 1020

ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc 1080

gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt 1140

gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa 1200

caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcggcggt 1260

cgggctgtaa cccccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg 1320

gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc 1380

aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc 1440

gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt 1500

tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga 1560

aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg 1620

gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc 1680

ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg 1740

cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg 1800

ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt 1860

ttggcaaaga att 1873

<210> 43

<211> 370

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 43

gggggaggct gctggtgaat attaaccaag gtcaccccag ttatcggagg agcaaacagg 60

ggctaagtcc acctcgagcc atggcgatgc tctaatctct ctagacaagg ttcatatttg 120

tatgggttac ttattctctc tttgttgact aagtcaataa tcagaatcag caggtttgca 180

gtcagattgg cagggataag cagcctagct caggagaagt gagtataaaa gccccaggct 240

gggagcagcc atcagctagc gccggcaaga ggtaagggtt taagggatgg ttggttggtg 300

gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac 360

cggtgccacc 370

<210> 44

<211> 913

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 44

ggcatcctaa aaaatattca gtggaaacgt aaaaacatta aagactgatt aaacatcgca 60

gcatgacaca gatttagcaa ctgagcataa ataatttgac tcggatactg ctccaaaatc 120

cgaagaggac caatttcttc caggaggaca actacctcgt cctctgcaga cccctctcct 180

cggcagctga aggagtgtgg ccaatctgcc tccacctccc cgcggacccc ctactctcag 240

gacctcctgc agcaccccaa actggaagtg gccgctgcag acccaaggac gaggggcacg 300

cgggagccgg cagccctagt ggagcggttg gagatgttga ggtgggaggg tcacccaggt 360

ggggtgaggc tggggtaggt agcggagtga acggcttccg aagctctggg ccgcccccag 420

gttggactaa gcaggcgctc tgtcttcgcc cccgcccagg gtgggcgtct cctgaggact 480

ccccgccaca cctgacccga gaccgcgcgc ccagcctaga acgcttcccc gacccagcgt 540

agggccgccg cgactggcgg gcgagggtcg gcgggaggcc tggcgaaccc gggggcggga 600

ccaggcgggc aaggcccggc tgccgcagcg ccgctctgcg cgaggcggct ccgccgcggc 660

ggagggatac ggcgcaccat atatatatcg cggggcgcag actcgcgctc cggcagtggt 720

gctgggagtg tcgtggacgc cgtgccgtta ctcgtagtca ggcggcggcg caggcggcgg 780

cggcggcata gcgcacagcg cgccttagca gcagcagcag cagcagcggc atcggaggta 840

cccccgccgt cgcagccccc gcgctggtgc agccaccctc gctccctctg ctcttcctcc 900

cttcgctcgc acc 913

<210> 45

<211> 137

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 45

aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga 60

gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg 120

cctttctctc cacaggt 137

<210> 46

<211> 380

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 46

ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 60

catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 120

acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 180

ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 240

aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 300

ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat 360

tagtcatcgc tattaccatg 380

<210> 47

<211> 1246

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 47

tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa 60

ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg 120

ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg 180

cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc 240

ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc 300

gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt 360

tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg 420

tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg 480

gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg 540

ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt 600

gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg 660

ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg 720

gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc 780

acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg 840

gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg 900

gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc 960

gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa 1020

tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga 1080

agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc 1140

gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg 1200

ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcgg 1246

<210> 48

<211> 1061

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 48

tagggaggtc ctgcacgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 60

cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 120

tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 180

agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 240

gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 300

agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc 360

ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat 420

gggggcgggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc 480

ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct 540

tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga 600

gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc 660

cggctctgac tgaccgcgtt actaaaacag gtaagtccgg cctccgcgcc gggttttggc 720

gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag 780

cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc 840

ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag 900

ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga 960

ttctgcggag ggatctccgt ggggcggtga acgccgatga tgcctctact aaccatgttc 1020

atgttttctt tttttttcta caggtcctgg gtgacgaaca g 1061

<210> 49

<211> 953

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 49

aattcggtac cctagttatt aatagtaatc aattacgggg tcattagttc atagcccata 60

tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 120

cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 180

ccattgacgt caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt 240

gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 300

ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 360

catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc 420

cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg 480

ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg 540

ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt 600

tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt 660

cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720

ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780

gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840

ttgaggggct ccgggagcta gagcctctgc taaccatgtt catgccttct tctttttcct 900

acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aag 953

<210> 50

<211> 541

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 50

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacagggc cctgtctcct cagcttcagg 420

caccaccact gacctgggac agtgaatcct ctaaggtaaa tataaaattt ttaagtgtat 480

aatgtgttaa actactgatt ctaattgttt ctctctttta gattccaacc tttggaactg 540

a 541

<210> 51

<211> 612

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 51

gccccctttt gcatccagtt tattcctaca tttgtcacac tgttaacagc ccaccccttc 60

caatgagacc agtggtatca gtgagttgtg gagatcagga aaagggctca agagaaaggc 120

agtcaaagcc ctttttctgt ccctgtccca gctgctttaa taagatctcc ataagagaag 180

agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta gtaaaacatg 240

taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata taaaaaaaat 300

ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg acagcagggg gctgtttcat 360

atactgatga cctctttata gccaaccttt gttcatggca gccagcatat gggcatatgt 420

tgccaaactc taaaccaaat actcattctg atgttttaaa tgatttgccc tcccatatgt 480

ccttccgagt gagagacaca aaaaattcca acacactatt gcaatgaaaa taaatttcct 540

ttattagcca gaagtcagat gctcaagggg cttcatgatg tccccataat ttttggcaga 600

gggaaaaaga tc 612

<210> 52

<211> 1347

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 52

caagtgcagc tggttcaaag cggagccgaa gtgaaaaagc ccggagctag cgtgaaggtg 60

tcctgcaagg ccagcggcca catcttcagc aactactgga tccagtgggt gcggcaggcc 120

cctggccaag gcctggaatg gatgggcgag atcctgccag gatctggcca caccgagtac 180

acagagaact tcaaggatag agtgaccatg accagagata cctccacaag caccgtgtac 240

atggaactga gcagcctgag aagcgaggac acagctgtgt actactgcgc cagatacttt 300

tttggctcat cccctaactg gtacttcgac gtgtggggcc aaggcaccct tgtcaccgtc 360

agcagcgcca gtactaaggg acccagcgtg ttcccactgg ccccatgcag cagaagcaca 420

tctgaaagca cagccgccct gggttgtctg gtcaaagact acttccccga acccgtgaca 480

gtgagctgga acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc cgtgctgcag 540

agctctggcc tgtattctct gagtagcgtg gtcaccgttc ctagctccaa cttcggcaca 600

cagacctaca cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga taagaccgtt 660

gagagaaagt gctgcgtgga atgcccacca tgtccagctc caccagtcgc cggcccttct 720

gttttcctgt tcccaccaaa gcccaaagac accctgatga tcagccggac ccctgaagtg 780

acatgtgtgg tggtggatgt gtcccaggag gatcctgagg tgcagtttaa ttggtacgtt 840

gacggagtgg aagttcataa tgccaagacc aaaccccgcg aggaacagtt taacagcacc 900

taccgggtgg tgtccgtgct gacagtgctg caccaggact ggctgaacgg caaagagtac 960

aagtgcaagg tgtctaacaa gggcctgcct agcagcatcg agaagaccat ctcaaaggcc 1020

aagggccagc ccagagagcc ccaagtctat acactgcctc cttctcaaga agaaatgaca 1080

aagaaccaag tgtctctgac ctgcctggtg aagggcttct accccagcga catcgccgtc 1140

gaatgggaga gcaacggaca gcctgaaaac aactacaaga cgacccctcc agtgctggac 1200

agcgatggca gcttcttcct gtattcacgg ctgaccgtgg acaagagccg atggcaagag 1260

ggcaacgtgt ttagctgcag cgtgctccac gaagccctgc acagccacta cacccagaag 1320

tccctgagcc tgtctctggg aaaataa 1347

<210> 53

<211> 642

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 53

gatatccaga tgacccagtc tccatctagc ctgtccgcca gcgtgggcga cagagtgacc 60

atcacctgcg gcgccagcga gaacatctat ggcgctctga actggtacca gcagaaacct 120

ggcaaggccc ctaagctgct gatctacggc gccaccaacc tggccgatgg cgtgcctagt 180

agattcagcg gatctggcag cggcacagac ttcaccctga ccatcagcag cctgcaacct 240

gaggactttg ccacatacta ctgccagaac gtgctgaata cacctctgac attcggccaa 300

ggaaccaaag tggaaatcaa gcggaccgtg gccgctccta gcgtgttcat cttccctcct 360

tccgatgaac aactgaagag cggaaccgcc tctgtggtgt gcctgctgaa caacttctac 420

cctagagagg ccaaggtgca gtggaaggtc gacaacgccc tgcagagcgg caacagccag 480

gagagcgtga cggaacagga cagcaaggac agcacctaca gcctgagctc cacccttaca 540

ctgtctaaag ccgactacga gaagcacaag gtgtacgcct gtgaagtgac acaccagggc 600

ctgagcagcc ctgtgaccaa gtcttttaac cggggcgagt gc 642

<210> 54

<211> 173

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 54

actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 60

aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg 120

gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgt 173

<210> 55

<211> 100

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 55

ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg 60

cggtaggcgt gtacggtggg aggtctatat aagcagagct 100

<210> 56

<211> 380

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 56

ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 60

catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 120

acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 180

ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 240

aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 300

ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat 360

tagtcatcgc tattaccatg 380

<210> 57

<211> 1246

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 57

tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa 60

ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg 120

ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg 180

cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc 240

ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc 300

gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt 360

tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg 420

tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg 480

gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg 540

ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt 600

gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg 660

ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg 720

gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc 780

acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg 840

gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg 900

gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc 960

gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa 1020

tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga 1080

agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc 1140

gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg 1200

ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcgg 1246

<210> 58

<211> 201

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 58

gaattcgggc ggagttaggg cggagccaat cagcgtgcgc cgttccgaaa gttgcctttt 60

atggctgggc ggagaatggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg 120

gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatccctgt 180

gatcgtgatc atcacttgtg a 201

<210> 59

<211> 953

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 59

aattcggtac cctagttatt aatagtaatc aattacgggg tcattagttc atagcccata 60

tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 120

cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 180

ccattgacgt caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt 240

gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 300

ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 360

catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc 420

cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg 480

ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg 540

ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt 600

tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt 660

cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720

ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780

gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840

ttgaggggct ccgggagcta gagcctctgc taaccatgtt catgccttct tctttttcct 900

acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aag 953

<210> 60

<211> 1766

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 60

gtaaatttta tggaatgtga atcataattc aatttttcaa catgcgttag gagggacatt 60

tcaaactctt ttttacccta gactttccta ccatcaccca gagtatccag ccaggagggg 120

aggggctaga gacaccagaa gtttagcagg gaggagggcg tagggattcg gggaatgaag 180

ggatgggatt cagactaggg ccaggaccca gggatggaga gaaagagatg agagtggttt 240

gggggcttgg tgacttagag aacagagctg caggctcaga ggcacacagg agtttctggg 300

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 360

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 420

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 480

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 540

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag ggcttaagcg 600

tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 660

gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa 720

agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 780

gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt 840

gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga 900

attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc 960

cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg 1020

gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc 1080

tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg 1140

taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg 1200

gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga 1260

gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc 1320

cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg 1380

aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg 1440

gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg 1500

cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct 1560

tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca 1620

ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat 1680

ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt 1740

tttttcttcc atttcaggtg tcgtga 1766

<210> 61

<211> 82

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 61

gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct 60

gcctgaaatc actttttttc ag 82

<210> 62

<211> 1404

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 62

atgggctggt cctgcatcat cctgttcctg gtggccaccg ccacaggcgt gcacagccaa 60

gtgcagctgg ttcaaagcgg agccgaagtg aaaaagcccg gagctagcgt gaaggtgtcc 120

tgcaaggcca gcggccacat cttcagcaac tactggatcc agtgggtgcg gcaggcccct 180

ggccaaggcc tggaatggat gggcgagatc ctgccaggat ctggccacac cgagtacaca 240

gagaacttca aggatagagt gaccatgacc agagatacct ccacaagcac cgtgtacatg 300

gaactgagca gcctgagaag cgaggacaca gctgtgtact actgcgccag atactttttt 360

ggctcatccc ctaactggta cttcgacgtg tggggccaag gcacccttgt caccgtcagc 420

agcgccagta ctaagggacc cagcgtgttc ccactggccc catgcagcag aagcacatct 480

gaaagcacag ccgccctggg ttgtctggtc aaagactact tccccgaacc cgtgacagtg 540

agctggaaca gcggcgccct gacaagcggc gtgcacacct tcccagccgt gctgcagagc 600

tctggcctgt attctctgag tagcgtggtc accgttccta gctccaactt cggcacacag 660

acctacacct gtaatgtgga ccacaagcct agcaacacca aagtggataa gaccgttgag 720

agaaagtgct gcgtggaatg cccaccatgt ccagctccac cagtcgccgg cccttctgtt 780

ttcctgttcc caccaaagcc caaagacacc ctgatgatca gccggacccc tgaagtgaca 840

tgtgtggtgg tggatgtgtc ccaggaggat cctgaggtgc agtttaattg gtacgttgac 900

ggagtggaag ttcataatgc caagaccaaa ccccgcgagg aacagtttaa cagcacctac 960

cgggtggtgt ccgtgctgac agtgctgcac caggactggc tgaacggcaa agagtacaag 1020

tgcaaggtgt ctaacaaggg cctgcctagc agcatcgaga agaccatctc aaaggccaag 1080

ggccagccca gagagcccca agtctataca ctgcctcctt ctcaagaaga aatgacaaag 1140

aaccaagtgt ctctgacctg cctggtgaag ggcttctacc ccagcgacat cgccgtcgaa 1200

tgggagagca acggacagcc tgaaaacaac tacaagacga cccctccagt gctggacagc 1260

gatggcagct tcttcctgta ttcacggctg accgtggaca agagccgatg gcaagagggc 1320

aacgtgttta gctgcagcgt gctccacgaa gccctgcaca gccactacac ccagaagtcc 1380

ctgagcctgt ctctgggaaa ataa 1404

<210> 63

<211> 708

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 63

atggacatga gggtccctgc tcagctgctg gggctcctgc tgctctggct cagcggtgcc 60

agatgtgata tccagatgac ccagtctcca tctagcctgt ccgccagcgt gggcgacaga 120

gtgaccatca cctgcggcgc cagcgagaac atctatggcg ctctgaactg gtaccagcag 180

aaacctggca aggcccctaa gctgctgatc tacggcgcca ccaacctggc cgatggcgtg 240

cctagtagat tcagcggatc tggcagcggc acagacttca ccctgaccat cagcagcctg 300

caacctgagg actttgccac atactactgc cagaacgtgc tgaatacacc tctgacattc 360

ggccaaggaa ccaaagtgga aatcaagcgg accgtggccg ctcctagcgt gttcatcttc 420

cctccttccg atgaacaact gaagagcgga accgcctctg tggtgtgcct gctgaacaac 480

ttctacccta gagaggccaa ggtgcagtgg aaggtcgaca acgccctgca gagcggcaac 540

agccaggaga gcgtgacgga acaggacagc aaggacagca cctacagcct gagctccacc 600

cttacactgt ctaaagccga ctacgagaag cacaaggtgt acgcctgtga agtgacacac 660

cagggcctga gcagccctgt gaccaagtct tttaaccggg gcgagtgc 708

<210> 64

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 64

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly His Ile Phe Ser Asn Tyr

20 25 30

Trp Ile Gln Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Glu Ile Leu Pro Gly Ser Gly His Thr Glu Tyr Thr Glu Asn Phe

50 55 60

Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Phe Phe Gly Ser Ser Pro Asn Trp Tyr Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys

210 215 220

Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro

260 265 270

Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala

420 425 430

Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 65

<211> 1061

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 65

tagggaggtc ctgcacgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 60

cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 120

tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 180

agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 240

gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 300

agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc 360

ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat 420

gggggcgggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc 480

ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct 540

tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga 600

gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc 660

cggctctgac tgaccgcgtt actaaaacag gtaagtccgg cctccgcgcc gggttttggc 720

gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag 780

cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc 840

ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag 900

ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga 960

ttctgcggag ggatctccgt ggggcggtga acgccgatga tgcctctact aaccatgttc 1020

atgttttctt tttttttcta caggtcctgg gtgacgaaca g 1061

<210> 66

<211> 170

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 66

cgatgctcta atctctctag acaaggttca tatttgtatg ggttacttat tctctctttg 60

ttgactaagt caataatcag aatcagcagg tttgcagtca gattggcagg gataagcagc 120

ctagctcagg agaagtgagt ataaaagccc caggctggga gcagccatca 170

<210> 67

<211> 254

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 67

gggggaggct gctggtgaat attaaccaag gtcaccccag ttatcggagg agcaaacagg 60

ggctaagtcc acctcgagcc atggcgatgc tctaatctct ctagacaagg ttcatatttg 120

tatgggttac ttattctctc tttgttgact aagtcaataa tcagaatcag caggtttgca 180

gtcagattgg cagggataag cagcctagct caggagaagt gagtataaaa gccccaggct 240

gggagcagcc atca 254

<210> 68

<211> 592

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 68

gtaaatttta tggaatgtga atcataattc aatttttcaa catgcgttag gagggacatt 60

tcaaactctt ttttacccta gactttccta ccatcaccca gagtatccag ccaggagggg 120

aggggctaga gacaccagaa gtttagcagg gaggagggcg tagggattcg gggaatgaag 180

ggatgggatt cagactaggg ccaggaccca gggatggaga gaaagagatg agagtggttt 240

gggggcttgg tgacttagag aacagagctg caggctcaga ggcacacagg agtttctggg 300

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 360

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 420

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 480

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 540

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gg 592

<210> 69

<211> 205

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 69

aatgactcct ttcggtaagt gcagtggaag ctgtacactg cccaggcaaa gcgtccgggc 60

agcgtaggcg ggcgactcag atcccagcca gtggacttag cccctgtttg ctcctccgat 120

aactggggtg accttggtta atattcacca gcagcctccc ccgttgcccc tctggatcca 180

ctgcttaaat acggacgagg acagg 205

<210> 70

<211> 423

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 70

gctctaaccc actctgatct cccagggcgg cagtaagtct tcagcatcag gcattttggg 60

gtgactcagt aaatggtaga tcttgctacc agtggaacag ccactaagga ttctgcagtg 120

agagcagagg gccagctaag tggtactctc ccagagactg tctgactcac gccaccccct 180

ccaccttgga cacaggacgc tgtggtttct gagccaggta caatgactcc tttcggtaag 240

tgcagtggaa gctgtacact gcccaggcaa agcgtccggg cagcgtaggc gggcgactca 300

gatcccagcc agtggactta gcccctgttt gctcctccga taactggggt gaccttggtt 360

aatattcacc agcagcctcc cccgttgccc ctctggatcc actgcttaaa tacggacgag 420

gac 423

<210> 71

<211> 72

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 71

gggggaggct gctggtgaat attaaccaag gtcaccccag ttatcggagg agcaaacagg 60

ggctaagtcc ac 72

<210> 72

<211> 913

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 72

tagggaggtc ctgcacagaa ggggaggagg gggcagcagc tgtctgacca ctgttggtct 60

tgcaacttgt gtccccaggt taatttttaa aaagcagtca aaagtccaag tggcccttgg 120

cagcatttac tctctctgtt tgctctggtt aataatctca ggagcacaaa cattcctgga 180

ggcaggagaa gaaatcaaca tcctggactt atcctctggg cctctcccca cccccaggat 240

tgtaactgaa atgcttcact ggtgctcctt ttgttttaag gcattggatc ttcatagcta 300

ctgatcgtgc ccaagcacac agtatctgca gcaaccactt aggcctccag gaatgtggtg 360

accattgacc ctaattcatt ccccttcatg gatcctatgt aaccatcctc caaaaagagc 420

tttcgcaaac tcaaataaac acaggaaagg aagaccttct tatctttgag agtatatgtt 480

tagccctata gctctaaccc actctgatct cccagggcgg cagtaagtct tcagcatcag 540

gcattttggg gtgactcagt aaatggtaga tcttgctacc agtggaacag ccactaagga 600

ttctgcagtg agagcagagg gccagctaag tggtactctc ccagagactg tctgactcac 660

gccaccccct ccaccttgga cacaggacgc tgtggtttct gagccaggta caatgactcc 720

tttcggtaag tgcagtggaa gctgtacact gcccaggcaa agcgtccggg cagcgtaggc 780

gggcgactca gatcccagcc agtggactta gcccctgttt gctcctccga taactggggt 840

gaccttggtt aatattcacc agcagcctcc cccgttgccc ctctggatcc actgcttaaa 900

tacggacgag gac 913

<210> 73

<211> 37

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 73

gtagataagt agcatggcgg gttaatcatt aactaca 37

<210> 74

<211> 46

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 74

ggaggggtgg agtcgtgacg tgaattacgt catagggtta gggagg 46

<210> 75

<211> 182

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 75

gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg 60

gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 120

cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc 180

aa 182

<210> 76

<211> 191

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 76

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag 180

ggttagggag g 191

<210> 77

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 77

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Ala

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Gly Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Asn Val Leu Asn Thr Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 78

<211> 467

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 78

Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly

1 5 10 15

Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys

20 25 30

Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly His Ile Phe

35 40 45

Ser Asn Tyr Trp Ile Gln Trp Val Arg Gln Ala Pro Gly Gln Gly Leu

50 55 60

Glu Trp Met Gly Glu Ile Leu Pro Gly Ser Gly His Thr Glu Tyr Thr

65 70 75 80

Glu Asn Phe Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser

85 90 95

Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val

100 105 110

Tyr Tyr Cys Ala Arg Tyr Phe Phe Gly Ser Ser Pro Asn Trp Tyr Phe

115 120 125

Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr

130 135 140

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser

145 150 155 160

Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu

165 170 175

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His

180 185 190

Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser

195 200 205

Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys

210 215 220

Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu

225 230 235 240

Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala

245 250 255

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

260 265 270

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln

275 280 285

Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val

290 295 300

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr

305 310 315 320

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

325 330 335

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile

340 345 350

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

355 360 365

Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser

370 375 380

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

385 390 395 400

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

405 410 415

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val

420 425 430

Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Leu

435 440 445

His Glu Ala Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

450 455 460

Leu Gly Lys

465

<210> 79

<211> 236

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 79

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Ser Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

20 25 30

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Gly Ala Ser

35 40 45

Glu Asn Ile Tyr Gly Ala Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys

50 55 60

Ala Pro Lys Leu Leu Ile Tyr Gly Ala Thr Asn Leu Ala Asp Gly Val

65 70 75 80

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

85 90 95

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Asn

100 105 110

Val Leu Asn Thr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

115 120 125

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

130 135 140

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

145 150 155 160

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

165 170 175

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

180 185 190

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

195 200 205

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

210 215 220

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 80

<211> 19

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 80

Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly

1 5 10 15

Val His Ser

<210> 81

<211> 22

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 81

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Ser Gly Ala Arg Cys

20

<210> 82

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> synthetic polypeptide

<400> 82

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ile Phe Ser Asn Tyr

20 25 30

Trp Ile Gln Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Glu Ile Leu Pro Gly Ser Gly Ser Thr Glu Tyr Thr Glu Asn Phe

50 55 60

Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Phe Phe Gly Ser Ser Pro Asn Trp Tyr Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys

210 215 220

Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro

260 265 270

Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 83

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 83

caagtgcagc tggttcaaag cggagccgaa gtgaaaaagc ccggagctag cgtgaaggtg 60

tcctgcaagg ccagcggcta tatcttcagc aactactgga tccagtgggt gcggcaggcc 120

cctggccaag gcctggaatg gatgggcgag atcctgccag gatctggctc taccgagtac 180

acagagaact tcaaggatag agtgaccatg accagagata cctccacaag caccgtgtac 240

atggaactga gcagcctgag aagcgaggac acagctgtgt actactgcgc cagatacttt 300

tttggctcat cccctaactg gtacttcgac gtgtggggcc aaggcaccct tgtcaccgtc 360

agcagcgcca gtactaaggg acccagcgtg ttcccactgg ccccatgcag cagaagcaca 420

tctgaaagca cagccgccct gggttgtctg gtcaaagact acttccccga acccgtgaca 480

gtgagctgga acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc cgtgctgcag 540

agctctggcc tgtattctct gagtagcgtg gtcaccgttc ctagctccaa cttcggcaca 600

cagacctaca cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga taagaccgtt 660

gagagaaagt gctgcgtgga atgcccacca tgtccagctc caccagtcgc cggcccttct 720

gttttcctgt tcccaccaaa gcccaaagac accctgatga tcagccggac ccctgaagtg 780

acatgtgtgg tggtggatgt gtcccaggag gatcctgagg tgcagtttaa ttggtacgtt 840

gacggagtgg aagttcataa tgccaagacc aaaccccgcg aggaacagtt taacagcacc 900

taccgggtgg tgtccgtgct gacagtgctg caccaggact ggctgaacgg caaagagtac 960

aagtgcaagg tgtctaacaa gggcctgcct agcagcatcg agaagaccat ctcaaaggcc 1020

aagggccagc ccagagagcc ccaagtctat acactgcctc cttctcaaga agaaatgaca 1080

aagaaccaag tgtctctgac ctgcctggtg aagggcttct accccagcga catcgccgtc 1140

gaatgggaga gcaacggaca gcctgaaaac aactacaaga cgacccctcc agtgctggac 1200

agcgatggca gcttcttcct gtattcacgg ctgaccgtgg acaagagccg atggcaagag 1260

ggcaacgtgt ttagctgcag cgtgatgcac gaagccctgc acaaccacta cacccagaag 1320

tccctgagcc tgtctctggg aaaa 1344

<210> 84

<211> 4181

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 84

gtaaatttta tggaatgtga atcataattc aatttttcaa catgcgttag gagggacatt 60

tcaaactctt ttttacccta gactttccta ccatcaccca gagtatccag ccaggagggg 120

aggggctaga gacaccagaa gtttagcagg gaggagggcg tagggattcg gggaatgaag 180

ggatgggatt cagactaggg ccaggaccca gggatggaga gaaagagatg agagtggttt 240

gggggcttgg tgacttagag aacagagctg caggctcaga ggcacacagg agtttctggg 300

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 360

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 420

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 480

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 540

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag ggcttaagcg 600

tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 660

gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa 720

agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 780

gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt 840

gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga 900

attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc 960

cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg 1020

gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc 1080

tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg 1140

taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg 1200

gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga 1260

gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc 1320

cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg 1380

aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg 1440

gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg 1500

cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct 1560

tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca 1620

ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat 1680

ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt 1740

tttttcttcc atttcaggtg tcgtgaacta gtgccaccat gggctggtcc tgcatcatcc 1800

tgttcctggt ggccaccgcc acaggcgtgc acagccaagt gcagctggtt caaagcggag 1860

ccgaagtgaa aaagcccgga gctagcgtga aggtgtcctg caaggccagc ggccacatct 1920

tcagcaacta ctggatccag tgggtgcggc aggcccctgg ccaaggcctg gaatggatgg 1980

gcgagatcct gccaggatct ggccacaccg agtacacaga gaacttcaag gatagagtga 2040

ccatgaccag agatacctcc acaagcaccg tgtacatgga actgagcagc ctgagaagcg 2100

aggacacagc tgtgtactac tgcgccagat acttttttgg ctcatcccct aactggtact 2160

tcgacgtgtg gggccaaggc acccttgtca ccgtcagcag cgccagtact aagggaccca 2220

gcgtgttccc actggcccca tgcagcagaa gcacatctga aagcacagcc gccctgggtt 2280

gtctggtcaa agactacttc cccgaacccg tgacagtgag ctggaacagc ggcgccctga 2340

caagcggcgt gcacaccttc ccagccgtgc tgcagagctc tggcctgtat tctctgagta 2400

gcgtggtcac cgttcctagc tccaacttcg gcacacagac ctacacctgt aatgtggacc 2460

acaagcctag caacaccaaa gtggataaga ccgttgagag aaagtgctgc gtggaatgcc 2520

caccatgtcc agctccacca gtcgccggcc cttctgtttt cctgttccca ccaaagccca 2580

aagacaccct gatgatcagc cggacccctg aagtgacatg tgtggtggtg gatgtgtccc 2640

aggaggatcc tgaggtgcag tttaattggt acgttgacgg agtggaagtt cataatgcca 2700

agaccaaacc ccgcgaggaa cagtttaaca gcacctaccg ggtggtgtcc gtgctgacag 2760

tgctgcacca ggactggctg aacggcaaag agtacaagtg caaggtgtct aacaagggcc 2820

tgcctagcag catcgagaag accatctcaa aggccaaggg ccagcccaga gagccccaag 2880

tctatacact gcctccttct caagaagaaa tgacaaagaa ccaagtgtct ctgacctgcc 2940

tggtgaaggg cttctacccc agcgacatcg ccgtcgaatg ggagagcaac ggacagcctg 3000

aaaacaacta caagacgacc cctccagtgc tggacagcga tggcagcttc ttcctgtatt 3060

cacggctgac cgtggacaag agccgatggc aagagggcaa cgtgtttagc tgcagcgtgc 3120

tccacgaagc cctgcacagc cactacaccc agaagtccct gagcctgtct ctgggaaaaa 3180

gaaagagaag aggcagcgga gaaggcagag gcagcctgct gacctgcggc gacgtcgaag 3240

agaaccccgg ccctatggac atgagggtcc ctgctcagct gctggggctc ctgctgctct 3300

ggctcagcgg tgccagatgt gatatccaga tgacccagtc tccatctagc ctgtccgcca 3360

gcgtgggcga cagagtgacc atcacctgcg gcgccagcga gaacatctat ggcgctctga 3420

actggtacca gcagaaacct ggcaaggccc ctaagctgct gatctacggc gccaccaacc 3480

tggccgatgg cgtgcctagt agattcagcg gatctggcag cggcacagac ttcaccctga 3540

ccatcagcag cctgcaacct gaggactttg ccacatacta ctgccagaac gtgctgaata 3600

cacctctgac attcggccaa ggaaccaaag tggaaatcaa gcggaccgtg gccgctccta 3660

gcgtgttcat cttccctcct tccgatgaac aactgaagag cggaaccgcc tctgtggtgt 3720

gcctgctgaa caacttctac cctagagagg ccaaggtgca gtggaaggtc gacaacgccc 3780

tgcagagcgg caacagccag gagagcgtga cggaacagga cagcaaggac agcacctaca 3840

gcctgagctc cacccttaca ctgtctaaag ccgactacga gaagcacaag gtgtacgcct 3900

gtgaagtgac acaccagggc ctgagcagcc ctgtgaccaa gtcttttaac cggggcgagt 3960

gctgaattcg aatcgtacct agggatccag acatgataag atacattgat gagtttggac 4020

aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg 4080

ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt 4140

ttatgtttca ggttcagggg gaggtgtggg aggtttttta a 4181

<210> 85

<211> 2956

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 85

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacagggc cctgtctcct cagcttcagg 420

caccaccact gacctgggac agtgaatcct ctaaggtaaa tataaaattt ttaagtgtat 480

aatgtgttaa actactgatt ctaattgttt ctctctttta gattccaacc tttggaactg 540

aactagtgcc accatgggct ggtcctgcat catcctgttc ctggtggcca ccgccacagg 600

cgtgcacagc caagtgcagc tggttcaaag cggagccgaa gtgaaaaagc ccggagctag 660

cgtgaaggtg tcctgcaagg ccagcggcca catcttcagc aactactgga tccagtgggt 720

gcggcaggcc cctggccaag gcctggaatg gatgggcgag atcctgccag gatctggcca 780

caccgagtac acagagaact tcaaggatag agtgaccatg accagagata cctccacaag 840

caccgtgtac atggaactga gcagcctgag aagcgaggac acagctgtgt actactgcgc 900

cagatacttt tttggctcat cccctaactg gtacttcgac gtgtggggcc aaggcaccct 960

tgtcaccgtc agcagcgcca gtactaaggg acccagcgtg ttcccactgg ccccatgcag 1020

cagaagcaca tctgaaagca cagccgccct gggttgtctg gtcaaagact acttccccga 1080

acccgtgaca gtgagctgga acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc 1140

cgtgctgcag agctctggcc tgtattctct gagtagcgtg gtcaccgttc ctagctccaa 1200

cttcggcaca cagacctaca cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga 1260

taagaccgtt gagagaaagt gctgcgtgga atgcccacca tgtccagctc caccagtcgc 1320

cggcccttct gttttcctgt tcccaccaaa gcccaaagac accctgatga tcagccggac 1380

ccctgaagtg acatgtgtgg tggtggatgt gtcccaggag gatcctgagg tgcagtttaa 1440

ttggtacgtt gacggagtgg aagttcataa tgccaagacc aaaccccgcg aggaacagtt 1500

taacagcacc taccgggtgg tgtccgtgct gacagtgctg caccaggact ggctgaacgg 1560

caaagagtac aagtgcaagg tgtctaacaa gggcctgcct agcagcatcg agaagaccat 1620

ctcaaaggcc aagggccagc ccagagagcc ccaagtctat acactgcctc cttctcaaga 1680

agaaatgaca aagaaccaag tgtctctgac ctgcctggtg aagggcttct accccagcga 1740

catcgccgtc gaatgggaga gcaacggaca gcctgaaaac aactacaaga cgacccctcc 1800

agtgctggac agcgatggca gcttcttcct gtattcacgg ctgaccgtgg acaagagccg 1860

atggcaagag ggcaacgtgt ttagctgcag cgtgctccac gaagccctgc acagccacta 1920

cacccagaag tccctgagcc tgtctctggg aaaaagaaag agaagaggca gcggagaagg 1980

cagaggcagc ctgctgacct gcggcgacgt cgaagagaac cccggcccta tggacatgag 2040

ggtccctgct cagctgctgg ggctcctgct gctctggctc agcggtgcca gatgtgatat 2100

ccagatgacc cagtctccat ctagcctgtc cgccagcgtg ggcgacagag tgaccatcac 2160

ctgcggcgcc agcgagaaca tctatggcgc tctgaactgg taccagcaga aacctggcaa 2220

ggcccctaag ctgctgatct acggcgccac caacctggcc gatggcgtgc ctagtagatt 2280

cagcggatct ggcagcggca cagacttcac cctgaccatc agcagcctgc aacctgagga 2340

ctttgccaca tactactgcc agaacgtgct gaatacacct ctgacattcg gccaaggaac 2400

caaagtggaa atcaagcgga ccgtggccgc tcctagcgtg ttcatcttcc ctccttccga 2460

tgaacaactg aagagcggaa ccgcctctgt ggtgtgcctg ctgaacaact tctaccctag 2520

agaggccaag gtgcagtgga aggtcgacaa cgccctgcag agcggcaaca gccaggagag 2580

cgtgacggaa caggacagca aggacagcac ctacagcctg agctccaccc ttacactgtc 2640

taaagccgac tacgagaagc acaaggtgta cgcctgtgaa gtgacacacc agggcctgag 2700

cagccctgtg accaagtctt ttaaccgggg cgagtgctga attcgaatcg tacctaggga 2760

tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa 2820

aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg 2880

caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt 2940

gtgggaggtt ttttaa 2956

<210> 86

<211> 3445

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 86

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacagggc cctgtctcct cagcttcagg 420

caccaccact gacctgggac agtgaatcct ctaaggtaaa tataaaattt ttaagtgtat 480

aatgtgttaa actactgatt ctaattgttt ctctctttta gattccaacc tttggaactg 540

aactagtgcc accatgggct ggtcctgcat catcctgttc ctggtggcca ccgccacagg 600

cgtgcacagc caagtgcagc tggttcaaag cggagccgaa gtgaaaaagc ccggagctag 660

cgtgaaggtg tcctgcaagg ccagcggcca catcttcagc aactactgga tccagtgggt 720

gcggcaggcc cctggccaag gcctggaatg gatgggcgag atcctgccag gatctggcca 780

caccgagtac acagagaact tcaaggatag agtgaccatg accagagata cctccacaag 840

caccgtgtac atggaactga gcagcctgag aagcgaggac acagctgtgt actactgcgc 900

cagatacttt tttggctcat cccctaactg gtacttcgac gtgtggggcc aaggcaccct 960

tgtcaccgtc agcagcgcca gtactaaggg acccagcgtg ttcccactgg ccccatgcag 1020

cagaagcaca tctgaaagca cagccgccct gggttgtctg gtcaaagact acttccccga 1080

acccgtgaca gtgagctgga acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc 1140

cgtgctgcag agctctggcc tgtattctct gagtagcgtg gtcaccgttc ctagctccaa 1200

cttcggcaca cagacctaca cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga 1260

taagaccgtt gagagaaagt gctgcgtgga atgcccacca tgtccagctc caccagtcgc 1320

cggcccttct gttttcctgt tcccaccaaa gcccaaagac accctgatga tcagccggac 1380

ccctgaagtg acatgtgtgg tggtggatgt gtcccaggag gatcctgagg tgcagtttaa 1440

ttggtacgtt gacggagtgg aagttcataa tgccaagacc aaaccccgcg aggaacagtt 1500

taacagcacc taccgggtgg tgtccgtgct gacagtgctg caccaggact ggctgaacgg 1560

caaagagtac aagtgcaagg tgtctaacaa gggcctgcct agcagcatcg agaagaccat 1620

ctcaaaggcc aagggccagc ccagagagcc ccaagtctat acactgcctc cttctcaaga 1680

agaaatgaca aagaaccaag tgtctctgac ctgcctggtg aagggcttct accccagcga 1740

catcgccgtc gaatgggaga gcaacggaca gcctgaaaac aactacaaga cgacccctcc 1800

agtgctggac agcgatggca gcttcttcct gtattcacgg ctgaccgtgg acaagagccg 1860

atggcaagag ggcaacgtgt ttagctgcag cgtgctccac gaagccctgc acagccacta 1920

cacccagaag tccctgagcc tgtctctggg aaaataaaac gttcctcgag gctgtgcctt 1980

ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 2040

ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 2100

gtcattctat tctggggggt ggggtggggc aggacagcaa ggtccggagg gggaggctgc 2160

tggtgaatat taaccaaggt caccccagtt atcggaggag caaacagggg ctaagtccac 2220

ctcgagccat ggcgatgctc taatctctct agacaaggtt catatttgta tgggttactt 2280

attctctctt tgttgactaa gtcaataatc agaatcagca ggtttgcagt cagattggca 2340

gggataagca gcctagctca ggagaagtga gtataaaagc cccaggctgg gagcagccat 2400

cagctagcgc cggcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt 2460

ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat 2520

ggacatgagg gtccctgctc agctgctggg gctcctgctg ctctggctca gcggtgccag 2580

atgtgatatc cagatgaccc agtctccatc tagcctgtcc gccagcgtgg gcgacagagt 2640

gaccatcacc tgcggcgcca gcgagaacat ctatggcgct ctgaactggt accagcagaa 2700

acctggcaag gcccctaagc tgctgatcta cggcgccacc aacctggccg atggcgtgcc 2760

tagtagattc agcggatctg gcagcggcac agacttcacc ctgaccatca gcagcctgca 2820

acctgaggac tttgccacat actactgcca gaacgtgctg aatacacctc tgacattcgg 2880

ccaaggaacc aaagtggaaa tcaagcggac cgtggccgct cctagcgtgt tcatcttccc 2940

tccttccgat gaacaactga agagcggaac cgcctctgtg gtgtgcctgc tgaacaactt 3000

ctaccctaga gaggccaagg tgcagtggaa ggtcgacaac gccctgcaga gcggcaacag 3060

ccaggagagc gtgacggaac aggacagcaa ggacagcacc tacagcctga gctccaccct 3120

tacactgtct aaagccgact acgagaagca caaggtgtac gcctgtgaag tgacacacca 3180

gggcctgagc agccctgtga ccaagtcttt taaccggggc gagtgctgaa ttcgaatcgt 3240

acctagggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat 3300

gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat 3360

tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca 3420

gggggaggtg tgggaggttt tttaa 3445

<210> 87

<211> 4075

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 87

ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg 60

atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg 120

aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gcctcgagga 180

acgttttatt ttcccagaga caggctcagg gacttctggg tgtagtggct gtgcagggct 240

tcgtggagca cgctgcagct aaacacgttg ccctcttgcc atcggctctt gtccacggtc 300

agccgtgaat acaggaagaa gctgccatcg ctgtccagca ctggaggggt cgtcttgtag 360

ttgttttcag gctgtccgtt gctctcccat tcgacggcga tgtcgctggg gtagaagccc 420

ttcaccaggc aggtcagaga cacttggttc tttgtcattt cttcttgaga aggaggcagt 480

gtatagactt ggggctctct gggctggccc ttggcctttg agatggtctt ctcgatgctg 540

ctaggcaggc ccttgttaga caccttgcac ttgtactctt tgccgttcag ccagtcctgg 600

tgcagcactg tcagcacgga caccacccgg taggtgctgt taaactgttc ctcgcggggt 660

ttggtcttgg cattatgaac ttccactccg tcaacgtacc aattaaactg cacctcagga 720

tcctcctggg acacatccac caccacacat gtcacttcag gggtccggct gatcatcagg 780

gtgtctttgg gctttggtgg gaacaggaaa acagaagggc cggcgactgg tggagctgga 840

catggtgggc attccacgca gcactttctc tcaacggtct tatccacttt ggtgttgcta 900

ggcttgtggt ccacattaca ggtgtaggtc tgtgtgccga agttggagct aggaacggtg 960

accacgctac tcagagaata caggccagag ctctgcagca cggctgggaa ggtgtgcacg 1020

ccgcttgtca gggcgccgct gttccagctc actgtcacgg gttcggggaa gtagtctttg 1080

accagacaac ccagggcggc tgtgctttca gatgtgcttc tgctgcatgg ggccagtggg 1140

aacacgctgg gtcccttagt actggcgctg ctgacggtga caagggtgcc ttggccccac 1200

acgtcgaagt accagttagg ggatgagcca aaaaagtatc tggcgcagta gtacacagct 1260

gtgtcctcgc ttctcaggct gctcagttcc atgtacacgg tgcttgtgga ggtatctctg 1320

gtcatggtca ctctatcctt gaagttctct gtgtactcgg tgtggccaga tcctggcagg 1380

atctcgccca tccattccag gccttggcca ggggcctgcc gcacccactg gatccagtag 1440

ttgctgaaga tgtggccgct ggccttgcag gacaccttca cgctagctcc gggctttttc 1500

acttcggctc cgctttgaac cagctgcact tggctgtgca cgcctgtggc ggtggccacc 1560

aggaacagga tgatgcagga ccagcccatg gtggcactag ttcagttcca aaggttggaa 1620

tctaaaagag agaaacaatt agaatcagta gtttaacaca ttatacactt aaaaatttta 1680

tatttacctt agaggattca ctgtcccagg tcagtggtgg tgcctgaagc tgaggagaca 1740

gggccctgtc ctcgtccgta tttaagcagt ggatccagag gggcaacggg ggaggctgct 1800

ggtgaatatt aaccaaggtc accccagtta tcggaggagc aaacaggggc taagtccact 1860

ggctgggatc tgagtcgccc gcctacgctg cccggacgct ttgcctgggc agtgtacagc 1920

ttccactgca cttaccgaaa ggagtcattc ccctctcaca ctacctaaac cacgccagga 1980

caacctctgc tcctctccac cgaaattcca aggggtcgag tggatgttgg aggtggcatg 2040

ggcccagaga ggtctctgac ctctgcccca gctccaaggt cagcaggcag ggagggctgt 2100

gtgtttgctg tttgctgctt gcaatgtttg cccattttag ggccgcggca cgtgcttaag 2160

gccccctttt gcatccagtt tattcctaca tttgtcacac tgttaacagc ccaccccttc 2220

caatgagacc agtggtatca gtgagttgtg gagatcagga aaagggctca agagaaaggc 2280

agtcaaagcc ctttttctgt ccctgtccca gctgctttaa taagatctcc ataagagaag 2340

agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta gtaaaacatg 2400

taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata taaaaaaaat 2460

ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg acagcagggg gctgtttcat 2520

atactgatga cctctttata gccaaccttt gttcatggca gccagcatat gggcatatgt 2580

tgccaaactc taaaccaaat actcattctg atgttttaaa tgatttgccc tcccatatgt 2640

ccttccgagt gagagacaca aaaaattcca acacactatt gcaatgaaaa taaatttcct 2700

ttattagcca gaagtcagat gctcaagggg cttcatgatg tccccataat ttttggcaga 2760

gggaaaaaga tctccggagg gggaggctgc tggtgaatat taaccaaggt caccccagtt 2820

atcggaggag caaacagggg ctaagtccac ctcgagccat ggcgatgctc taatctctct 2880

agacaaggtt catatttgta tgggttactt attctctctt tgttgactaa gtcaataatc 2940

agaatcagca ggtttgcagt cagattggca gggataagca gcctagctca ggagaagtga 3000

gtataaaagc cccaggctgg gagcagccat cagctagcgc cggcaagagg taagggttta 3060

agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca 3120

ctttttttca ggttggaccg gtgccaccat ggacatgagg gtccctgctc agctgctggg 3180

gctcctgctg ctctggctca gcggtgccag atgtgatatc cagatgaccc agtctccatc 3240

tagcctgtcc gccagcgtgg gcgacagagt gaccatcacc tgcggcgcca gcgagaacat 3300

ctatggcgct ctgaactggt accagcagaa acctggcaag gcccctaagc tgctgatcta 3360

cggcgccacc aacctggccg atggcgtgcc tagtagattc agcggatctg gcagcggcac 3420

agacttcacc ctgaccatca gcagcctgca acctgaggac tttgccacat actactgcca 3480

gaacgtgctg aatacacctc tgacattcgg ccaaggaacc aaagtggaaa tcaagcggac 3540

cgtggccgct cctagcgtgt tcatcttccc tccttccgat gaacaactga agagcggaac 3600

cgcctctgtg gtgtgcctgc tgaacaactt ctaccctaga gaggccaagg tgcagtggaa 3660

ggtcgacaac gccctgcaga gcggcaacag ccaggagagc gtgacggaac aggacagcaa 3720

ggacagcacc tacagcctga gctccaccct tacactgtct aaagccgact acgagaagca 3780

caaggtgtac gcctgtgaag tgacacacca gggcctgagc agccctgtga ccaagtcttt 3840

taaccggggc gagtgctgaa ttcgaatcgt acctagggat ccagacatga taagatacat 3900

tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat 3960

ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa 4020

caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaa 4075

<210> 88

<211> 4588

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 88

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag 180

ggttagggag gtcctgcata tgcggccgcg taaattttat ggaatgtgaa tcataattca 240

atttttcaac atgcgttagg agggacattt caaactcttt tttaccctag actttcctac 300

catcacccag agtatccagc caggagggga ggggctagag acaccagaag tttagcaggg 360

aggagggcgt agggattcgg ggaatgaagg gatgggattc agactagggc caggacccag 420

ggatggagag aaagagatga gagtggtttg ggggcttggt gacttagaga acagagctgc 480

aggctcagag gcacacagga gtttctgggc tcaccctgcc cccttccaac ccctcagttc 540

ccatcctcca gcagctgttt gtgtgctgcc tctgaagtcc acactgaaca aacttcagcc 600

tactcatgtc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca cacagccctc 660

cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc ccatgccacc 720

tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt gtcctggcgt 780

ggtttaggta gtgtgagagg gcttaagcgt gaggctccgg tgcccgtcag tgggcagagc 840

gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct 900

agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc 960

ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca 1020

acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct 1080

ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc 1140

ttgatcccga gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc 1200

ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg 1260

cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc 1320

gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat 1380

ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc 1440

gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg 1500

gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct 1560

ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg 1620

gggctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag 1680

gaaaggggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc 1740

gtccaggcac ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga 1800

ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc 1860

ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat 1920

tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaactag 1980

tgccaccatg ggctggtcct gcatcatcct gttcctggtg gccaccgcca caggcgtgca 2040

cagccaagtg cagctggttc aaagcggagc cgaagtgaaa aagcccggag ctagcgtgaa 2100

ggtgtcctgc aaggccagcg gccacatctt cagcaactac tggatccagt gggtgcggca 2160

ggcccctggc caaggcctgg aatggatggg cgagatcctg ccaggatctg gccacaccga 2220

gtacacagag aacttcaagg atagagtgac catgaccaga gatacctcca caagcaccgt 2280

gtacatggaa ctgagcagcc tgagaagcga ggacacagct gtgtactact gcgccagata 2340

cttttttggc tcatccccta actggtactt cgacgtgtgg ggccaaggca cccttgtcac 2400

cgtcagcagc gccagtacta agggacccag cgtgttccca ctggccccat gcagcagaag 2460

cacatctgaa agcacagccg ccctgggttg tctggtcaaa gactacttcc ccgaacccgt 2520

gacagtgagc tggaacagcg gcgccctgac aagcggcgtg cacaccttcc cagccgtgct 2580

gcagagctct ggcctgtatt ctctgagtag cgtggtcacc gttcctagct ccaacttcgg 2640

cacacagacc tacacctgta atgtggacca caagcctagc aacaccaaag tggataagac 2700

cgttgagaga aagtgctgcg tggaatgccc accatgtcca gctccaccag tcgccggccc 2760

ttctgttttc ctgttcccac caaagcccaa agacaccctg atgatcagcc ggacccctga 2820

agtgacatgt gtggtggtgg atgtgtccca ggaggatcct gaggtgcagt ttaattggta 2880

cgttgacgga gtggaagttc ataatgccaa gaccaaaccc cgcgaggaac agtttaacag 2940

cacctaccgg gtggtgtccg tgctgacagt gctgcaccag gactggctga acggcaaaga 3000

gtacaagtgc aaggtgtcta acaagggcct gcctagcagc atcgagaaga ccatctcaaa 3060

ggccaagggc cagcccagag agccccaagt ctatacactg cctccttctc aagaagaaat 3120

gacaaagaac caagtgtctc tgacctgcct ggtgaagggc ttctacccca gcgacatcgc 3180

cgtcgaatgg gagagcaacg gacagcctga aaacaactac aagacgaccc ctccagtgct 3240

ggacagcgat ggcagcttct tcctgtattc acggctgacc gtggacaaga gccgatggca 3300

agagggcaac gtgtttagct gcagcgtgct ccacgaagcc ctgcacagcc actacaccca 3360

gaagtccctg agcctgtctc tgggaaaaag aaagagaaga ggcagcggag aaggcagagg 3420

cagcctgctg acctgcggcg acgtcgaaga gaaccccggc cctatggaca tgagggtccc 3480

tgctcagctg ctggggctcc tgctgctctg gctcagcggt gccagatgtg atatccagat 3540

gacccagtct ccatctagcc tgtccgccag cgtgggcgac agagtgacca tcacctgcgg 3600

cgccagcgag aacatctatg gcgctctgaa ctggtaccag cagaaacctg gcaaggcccc 3660

taagctgctg atctacggcg ccaccaacct ggccgatggc gtgcctagta gattcagcgg 3720

atctggcagc ggcacagact tcaccctgac catcagcagc ctgcaacctg aggactttgc 3780

cacatactac tgccagaacg tgctgaatac acctctgaca ttcggccaag gaaccaaagt 3840

ggaaatcaag cggaccgtgg ccgctcctag cgtgttcatc ttccctcctt ccgatgaaca 3900

actgaagagc ggaaccgcct ctgtggtgtg cctgctgaac aacttctacc ctagagaggc 3960

caaggtgcag tggaaggtcg acaacgccct gcagagcggc aacagccagg agagcgtgac 4020

ggaacaggac agcaaggaca gcacctacag cctgagctcc acccttacac tgtctaaagc 4080

cgactacgag aagcacaagg tgtacgcctg tgaagtgaca caccagggcc tgagcagccc 4140

tgtgaccaag tcttttaacc ggggcgagtg ctgaattcga atcgtaccta gggatccaga 4200

catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 4260

ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 4320

acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga 4380

ggttttttaa gcttgtttaa acgtacgtag ataagtagca tggcgggtta atcattaact 4440

acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg 4500

aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg 4560

agcgagcgcg cagagaggga gtggccaa 4588

<210> 89

<211> 3375

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 89

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag 180

ggttagggag gtcctgcata tgcggccgca cgtgccgcgg ccctaaaatg ggcaaacatt 240

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 300

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 360

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gggaatgact 420

cctttcggta agtgcagtgg aagctgtaca ctgcccaggc aaagcgtccg ggcagcgtag 480

gcgggcgact cagatcccag ccagtggact tagcccctgt ttgctcctcc gataactggg 540

gtgaccttgg ttaatattca ccagcagcct cccccgttgc ccctctggat ccactgctta 600

aatacggacg aggacagggc cctgtctcct cagcttcagg caccaccact gacctgggac 660

agtgaatcct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt 720

ctaattgttt ctctctttta gattccaacc tttggaactg aactagtgcc accatgggct 780

ggtcctgcat catcctgttc ctggtggcca ccgccacagg cgtgcacagc caagtgcagc 840

tggttcaaag cggagccgaa gtgaaaaagc ccggagctag cgtgaaggtg tcctgcaagg 900

ccagcggcca catcttcagc aactactgga tccagtgggt gcggcaggcc cctggccaag 960

gcctggaatg gatgggcgag atcctgccag gatctggcca caccgagtac acagagaact 1020

tcaaggatag agtgaccatg accagagata cctccacaag caccgtgtac atggaactga 1080

gcagcctgag aagcgaggac acagctgtgt actactgcgc cagatacttt tttggctcat 1140

cccctaactg gtacttcgac gtgtggggcc aaggcaccct tgtcaccgtc agcagcgcca 1200

gtactaaggg acccagcgtg ttcccactgg ccccatgcag cagaagcaca tctgaaagca 1260

cagccgccct gggttgtctg gtcaaagact acttccccga acccgtgaca gtgagctgga 1320

acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc cgtgctgcag agctctggcc 1380

tgtattctct gagtagcgtg gtcaccgttc ctagctccaa cttcggcaca cagacctaca 1440

cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga taagaccgtt gagagaaagt 1500

gctgcgtgga atgcccacca tgtccagctc caccagtcgc cggcccttct gttttcctgt 1560

tcccaccaaa gcccaaagac accctgatga tcagccggac ccctgaagtg acatgtgtgg 1620

tggtggatgt gtcccaggag gatcctgagg tgcagtttaa ttggtacgtt gacggagtgg 1680

aagttcataa tgccaagacc aaaccccgcg aggaacagtt taacagcacc taccgggtgg 1740

tgtccgtgct gacagtgctg caccaggact ggctgaacgg caaagagtac aagtgcaagg 1800

tgtctaacaa gggcctgcct agcagcatcg agaagaccat ctcaaaggcc aagggccagc 1860

ccagagagcc ccaagtctat acactgcctc cttctcaaga agaaatgaca aagaaccaag 1920

tgtctctgac ctgcctggtg aagggcttct accccagcga catcgccgtc gaatgggaga 1980

gcaacggaca gcctgaaaac aactacaaga cgacccctcc agtgctggac agcgatggca 2040

gcttcttcct gtattcacgg ctgaccgtgg acaagagccg atggcaagag ggcaacgtgt 2100

ttagctgcag cgtgctccac gaagccctgc acagccacta cacccagaag tccctgagcc 2160

tgtctctggg aaaaagaaag agaagaggca gcggagaagg cagaggcagc ctgctgacct 2220

gcggcgacgt cgaagagaac cccggcccta tggacatgag ggtccctgct cagctgctgg 2280

ggctcctgct gctctggctc agcggtgcca gatgtgatat ccagatgacc cagtctccat 2340

ctagcctgtc cgccagcgtg ggcgacagag tgaccatcac ctgcggcgcc agcgagaaca 2400

tctatggcgc tctgaactgg taccagcaga aacctggcaa ggcccctaag ctgctgatct 2460

acggcgccac caacctggcc gatggcgtgc ctagtagatt cagcggatct ggcagcggca 2520

cagacttcac cctgaccatc agcagcctgc aacctgagga ctttgccaca tactactgcc 2580

agaacgtgct gaatacacct ctgacattcg gccaaggaac caaagtggaa atcaagcgga 2640

ccgtggccgc tcctagcgtg ttcatcttcc ctccttccga tgaacaactg aagagcggaa 2700

ccgcctctgt ggtgtgcctg ctgaacaact tctaccctag agaggccaag gtgcagtgga 2760

aggtcgacaa cgccctgcag agcggcaaca gccaggagag cgtgacggaa caggacagca 2820

aggacagcac ctacagcctg agctccaccc ttacactgtc taaagccgac tacgagaagc 2880

acaaggtgta cgcctgtgaa gtgacacacc agggcctgag cagccctgtg accaagtctt 2940

ttaaccgggg cgagtgctga attcgaatcg tacctaggga tccagacatg ataagataca 3000

ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 3060

tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca 3120

acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaacctg 3180

caggtctaga tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct 3240

agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc 3300

aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag 3360

agagggagtg gccaa 3375

<210> 90

<211> 3863

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 90

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag 180

ggttagggag gtcctgcata tgcggccgca cgtgccgcgg ccctaaaatg ggcaaacatt 240

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 300

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 360

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gggaatgact 420

cctttcggta agtgcagtgg aagctgtaca ctgcccaggc aaagcgtccg ggcagcgtag 480

gcgggcgact cagatcccag ccagtggact tagcccctgt ttgctcctcc gataactggg 540

gtgaccttgg ttaatattca ccagcagcct cccccgttgc ccctctggat ccactgctta 600

aatacggacg aggacagggc cctgtctcct cagcttcagg caccaccact gacctgggac 660

agtgaatcct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt 720

ctaattgttt ctctctttta gattccaacc tttggaactg aactagtgcc accatgggct 780

ggtcctgcat catcctgttc ctggtggcca ccgccacagg cgtgcacagc caagtgcagc 840

tggttcaaag cggagccgaa gtgaaaaagc ccggagctag cgtgaaggtg tcctgcaagg 900

ccagcggcca catcttcagc aactactgga tccagtgggt gcggcaggcc cctggccaag 960

gcctggaatg gatgggcgag atcctgccag gatctggcca caccgagtac acagagaact 1020

tcaaggatag agtgaccatg accagagata cctccacaag caccgtgtac atggaactga 1080

gcagcctgag aagcgaggac acagctgtgt actactgcgc cagatacttt tttggctcat 1140

cccctaactg gtacttcgac gtgtggggcc aaggcaccct tgtcaccgtc agcagcgcca 1200

gtactaaggg acccagcgtg ttcccactgg ccccatgcag cagaagcaca tctgaaagca 1260

cagccgccct gggttgtctg gtcaaagact acttccccga acccgtgaca gtgagctgga 1320

acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc cgtgctgcag agctctggcc 1380

tgtattctct gagtagcgtg gtcaccgttc ctagctccaa cttcggcaca cagacctaca 1440

cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga taagaccgtt gagagaaagt 1500

gctgcgtgga atgcccacca tgtccagctc caccagtcgc cggcccttct gttttcctgt 1560

tcccaccaaa gcccaaagac accctgatga tcagccggac ccctgaagtg acatgtgtgg 1620

tggtggatgt gtcccaggag gatcctgagg tgcagtttaa ttggtacgtt gacggagtgg 1680

aagttcataa tgccaagacc aaaccccgcg aggaacagtt taacagcacc taccgggtgg 1740

tgtccgtgct gacagtgctg caccaggact ggctgaacgg caaagagtac aagtgcaagg 1800

tgtctaacaa gggcctgcct agcagcatcg agaagaccat ctcaaaggcc aagggccagc 1860

ccagagagcc ccaagtctat acactgcctc cttctcaaga agaaatgaca aagaaccaag 1920

tgtctctgac ctgcctggtg aagggcttct accccagcga catcgccgtc gaatgggaga 1980

gcaacggaca gcctgaaaac aactacaaga cgacccctcc agtgctggac agcgatggca 2040

gcttcttcct gtattcacgg ctgaccgtgg acaagagccg atggcaagag ggcaacgtgt 2100

ttagctgcag cgtgctccac gaagccctgc acagccacta cacccagaag tccctgagcc 2160

tgtctctggg aaaataaaac gttcctcgag gctgtgcctt ctagttgcca gccatctgtt 2220

gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2280

taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 2340

ggggtggggc aggacagcaa ggtccggagg gggaggctgc tggtgaatat taaccaaggt 2400

caccccagtt atcggaggag caaacagggg ctaagtccac ctcgagccat ggcgatgctc 2460

taatctctct agacaaggtt catatttgta tgggttactt attctctctt tgttgactaa 2520

gtcaataatc agaatcagca ggtttgcagt cagattggca gggataagca gcctagctca 2580

ggagaagtga gtataaaagc cccaggctgg gagcagccat cagctagcgc cggcaagagg 2640

taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg 2700

cctgaaatca ctttttttca ggttggaccg gtgccaccat ggacatgagg gtccctgctc 2760

agctgctggg gctcctgctg ctctggctca gcggtgccag atgtgatatc cagatgaccc 2820

agtctccatc tagcctgtcc gccagcgtgg gcgacagagt gaccatcacc tgcggcgcca 2880

gcgagaacat ctatggcgct ctgaactggt accagcagaa acctggcaag gcccctaagc 2940

tgctgatcta cggcgccacc aacctggccg atggcgtgcc tagtagattc agcggatctg 3000

gcagcggcac agacttcacc ctgaccatca gcagcctgca acctgaggac tttgccacat 3060

actactgcca gaacgtgctg aatacacctc tgacattcgg ccaaggaacc aaagtggaaa 3120

tcaagcggac cgtggccgct cctagcgtgt tcatcttccc tccttccgat gaacaactga 3180

agagcggaac cgcctctgtg gtgtgcctgc tgaacaactt ctaccctaga gaggccaagg 3240

tgcagtggaa ggtcgacaac gccctgcaga gcggcaacag ccaggagagc gtgacggaac 3300

aggacagcaa ggacagcacc tacagcctga gctccaccct tacactgtct aaagccgact 3360

acgagaagca caaggtgtac gcctgtgaag tgacacacca gggcctgagc agccctgtga 3420

ccaagtcttt taaccggggc gagtgctgaa ttcgaatcgt acctagggat ccagacatga 3480

taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 3540

tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 3600

ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt 3660

tttaagcttg tttaaacgta cgtagataag tagcatggcg ggttaatcat taactacaag 3720

gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc 3780

gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga 3840

gcgcgcagag agggagtggc caa 3863

<210> 91

<211> 4573

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 91

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag 180

ggttagggag gtcctgcata tgcggccgcg tcatagggtt agggaggtcc tgcacacgtg 240

acgcgtcctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac 300

aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg 360

tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagcct 420

cgaggaacgt tttattttcc cagagacagg ctcagggact tctgggtgta gtggctgtgc 480

agggcttcgt ggagcacgct gcagctaaac acgttgccct cttgccatcg gctcttgtcc 540

acggtcagcc gtgaatacag gaagaagctg ccatcgctgt ccagcactgg aggggtcgtc 600

ttgtagttgt tttcaggctg tccgttgctc tcccattcga cggcgatgtc gctggggtag 660

aagcccttca ccaggcaggt cagagacact tggttctttg tcatttcttc ttgagaagga 720

ggcagtgtat agacttgggg ctctctgggc tggcccttgg cctttgagat ggtcttctcg 780

atgctgctag gcaggccctt gttagacacc ttgcacttgt actctttgcc gttcagccag 840

tcctggtgca gcactgtcag cacggacacc acccggtagg tgctgttaaa ctgttcctcg 900

cggggtttgg tcttggcatt atgaacttcc actccgtcaa cgtaccaatt aaactgcacc 960

tcaggatcct cctgggacac atccaccacc acacatgtca cttcaggggt ccggctgatc 1020

atcagggtgt ctttgggctt tggtgggaac aggaaaacag aagggccggc gactggtgga 1080

gctggacatg gtgggcattc cacgcagcac tttctctcaa cggtcttatc cactttggtg 1140

ttgctaggct tgtggtccac attacaggtg taggtctgtg tgccgaagtt ggagctagga 1200

acggtgacca cgctactcag agaatacagg ccagagctct gcagcacggc tgggaaggtg 1260

tgcacgccgc ttgtcagggc gccgctgttc cagctcactg tcacgggttc ggggaagtag 1320

tctttgacca gacaacccag ggcggctgtg ctttcagatg tgcttctgct gcatggggcc 1380

agtgggaaca cgctgggtcc cttagtactg gcgctgctga cggtgacaag ggtgccttgg 1440

ccccacacgt cgaagtacca gttaggggat gagccaaaaa agtatctggc gcagtagtac 1500

acagctgtgt cctcgcttct caggctgctc agttccatgt acacggtgct tgtggaggta 1560

tctctggtca tggtcactct atccttgaag ttctctgtgt actcggtgtg gccagatcct 1620

ggcaggatct cgcccatcca ttccaggcct tggccagggg cctgccgcac ccactggatc 1680

cagtagttgc tgaagatgtg gccgctggcc ttgcaggaca ccttcacgct agctccgggc 1740

tttttcactt cggctccgct ttgaaccagc tgcacttggc tgtgcacgcc tgtggcggtg 1800

gccaccagga acaggatgat gcaggaccag cccatggtgg cactagttca gttccaaagg 1860

ttggaatcta aaagagagaa acaattagaa tcagtagttt aacacattat acacttaaaa 1920

attttatatt taccttagag gattcactgt cccaggtcag tggtggtgcc tgaagctgag 1980

gagacagggc cctgtcctcg tccgtattta agcagtggat ccagaggggc aacgggggag 2040

gctgctggtg aatattaacc aaggtcaccc cagttatcgg aggagcaaac aggggctaag 2100

tccactggct gggatctgag tcgcccgcct acgctgcccg gacgctttgc ctgggcagtg 2160

tacagcttcc actgcactta ccgaaaggag tcattcccct ctcacactac ctaaaccacg 2220

ccaggacaac ctctgctcct ctccaccgaa attccaaggg gtcgagtgga tgttggaggt 2280

ggcatgggcc cagagaggtc tctgacctct gccccagctc caaggtcagc aggcagggag 2340

ggctgtgtgt ttgctgtttg ctgcttgcaa tgtttgccca ttttagggcc gcggcacgtg 2400

cttaaggccc ccttttgcat ccagtttatt cctacatttg tcacactgtt aacagcccac 2460

cccttccaat gagaccagtg gtatcagtga gttgtggaga tcaggaaaag ggctcaagag 2520

aaaggcagtc aaagcccttt ttctgtccct gtcccagctg ctttaataag atctccataa 2580

gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa 2640

aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatataaa 2700

aaaaatctaa cctcaagtca aggcttttct atggaataag gaatggacag cagggggctg 2760

tttcatatac tgatgacctc tttatagcca acctttgttc atggcagcca gcatatgggc 2820

atatgttgcc aaactctaaa ccaaatactc attctgatgt tttaaatgat ttgccctccc 2880

atatgtcctt ccgagtgaga gacacaaaaa attccaacac actattgcaa tgaaaataaa 2940

tttcctttat tagccagaag tcagatgctc aaggggcttc atgatgtccc cataattttt 3000

ggcagaggga aaaagatctc cggaggggga ggctgctggt gaatattaac caaggtcacc 3060

ccagttatcg gaggagcaaa caggggctaa gtccacctcg agccatggcg atgctctaat 3120

ctctctagac aaggttcata tttgtatggg ttacttattc tctctttgtt gactaagtca 3180

ataatcagaa tcagcaggtt tgcagtcaga ttggcaggga taagcagcct agctcaggag 3240

aagtgagtat aaaagcccca ggctgggagc agccatcagc tagcgccggc aagaggtaag 3300

ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg 3360

aaatcacttt ttttcaggtt ggaccggtgc caccatggac atgagggtcc ctgctcagct 3420

gctggggctc ctgctgctct ggctcagcgg tgccagatgt gatatccaga tgacccagtc 3480

tccatctagc ctgtccgcca gcgtgggcga cagagtgacc atcacctgcg gcgccagcga 3540

gaacatctat ggcgctctga actggtacca gcagaaacct ggcaaggccc ctaagctgct 3600

gatctacggc gccaccaacc tggccgatgg cgtgcctagt agattcagcg gatctggcag 3660

cggcacagac ttcaccctga ccatcagcag cctgcaacct gaggactttg ccacatacta 3720

ctgccagaac gtgctgaata cacctctgac attcggccaa ggaaccaaag tggaaatcaa 3780

gcggaccgtg gccgctccta gcgtgttcat cttccctcct tccgatgaac aactgaagag 3840

cggaaccgcc tctgtggtgt gcctgctgaa caacttctac cctagagagg ccaaggtgca 3900

gtggaaggtc gacaacgccc tgcagagcgg caacagccag gagagcgtga cggaacagga 3960

cagcaaggac agcacctaca gcctgagctc cacccttaca ctgtctaaag ccgactacga 4020

gaagcacaag gtgtacgcct gtgaagtgac acaccagggc ctgagcagcc ctgtgaccaa 4080

gtcttttaac cggggcgagt gctgaattcg aatcgtacct agggatccag acatgataag 4140

atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg 4200

tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa 4260

caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta 4320

agcttgttta aacgtacgta gataagtagc atggcgggtt aatcattaac tacacctgca 4380

ggtctagata cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag 4440

tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 4500

aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag 4560

agggagtggc caa 4573

<210> 92

<211> 2142

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 92

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacagggc cctgtctcct cagcttcagg 420

caccaccact gacctgggac agtgaatcct ctaaggtaaa tataaaattt ttaagtgtat 480

aatgtgttaa actactgatt ctaattgttt ctctctttta gattccaacc tttggaactg 540

aactagtgcc accatgggct ggtcctgcat catcctgttc ctggtggcca ccgccacagg 600

cgtgcacagc caagtgcagc tggttcaaag cggagccgaa gtgaaaaagc ccggagctag 660

cgtgaaggtg tcctgcaagg ccagcggcca catcttcagc aactactgga tccagtgggt 720

gcggcaggcc cctggccaag gcctggaatg gatgggcgag atcctgccag gatctggcca 780

caccgagtac acagagaact tcaaggatag agtgaccatg accagagata cctccacaag 840

caccgtgtac atggaactga gcagcctgag aagcgaggac acagctgtgt actactgcgc 900

cagatacttt tttggctcat cccctaactg gtacttcgac gtgtggggcc aaggcaccct 960

tgtcaccgtc agcagcgcca gtactaaggg acccagcgtg ttcccactgg ccccatgcag 1020

cagaagcaca tctgaaagca cagccgccct gggttgtctg gtcaaagact acttccccga 1080

acccgtgaca gtgagctgga acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc 1140

cgtgctgcag agctctggcc tgtattctct gagtagcgtg gtcaccgttc ctagctccaa 1200

cttcggcaca cagacctaca cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga 1260

taagaccgtt gagagaaagt gctgcgtgga atgcccacca tgtccagctc caccagtcgc 1320

cggcccttct gttttcctgt tcccaccaaa gcccaaagac accctgatga tcagccggac 1380

ccctgaagtg acatgtgtgg tggtggatgt gtcccaggag gatcctgagg tgcagtttaa 1440

ttggtacgtt gacggagtgg aagttcataa tgccaagacc aaaccccgcg aggaacagtt 1500

taacagcacc taccgggtgg tgtccgtgct gacagtgctg caccaggact ggctgaacgg 1560

caaagagtac aagtgcaagg tgtctaacaa gggcctgcct agcagcatcg agaagaccat 1620

ctcaaaggcc aagggccagc ccagagagcc ccaagtctat acactgcctc cttctcaaga 1680

agaaatgaca aagaaccaag tgtctctgac ctgcctggtg aagggcttct accccagcga 1740

catcgccgtc gaatgggaga gcaacggaca gcctgaaaac aactacaaga cgacccctcc 1800

agtgctggac agcgatggca gcttcttcct gtattcacgg ctgaccgtgg acaagagccg 1860

atggcaagag ggcaacgtgt ttagctgcag cgtgctccac gaagccctgc acagccacta 1920

cacccagaag tccctgagcc tgtctctggg aaaataaaac gttcctcgag gctgtgcctt 1980

ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 2040

ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 2100

gtcattctat tctggggggt ggggtggggc aggacagcaa gg 2142

<210> 93

<211> 1297

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 93

gggggaggct gctggtgaat attaaccaag gtcaccccag ttatcggagg agcaaacagg 60

ggctaagtcc acctcgagcc atggcgatgc tctaatctct ctagacaagg ttcatatttg 120

tatgggttac ttattctctc tttgttgact aagtcaataa tcagaatcag caggtttgca 180

gtcagattgg cagggataag cagcctagct caggagaagt gagtataaaa gccccaggct 240

gggagcagcc atcagctagc gccggcaaga ggtaagggtt taagggatgg ttggttggtg 300

gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac 360

cggtgccacc atggacatga gggtccctgc tcagctgctg gggctcctgc tgctctggct 420

cagcggtgcc agatgtgata tccagatgac ccagtctcca tctagcctgt ccgccagcgt 480

gggcgacaga gtgaccatca cctgcggcgc cagcgagaac atctatggcg ctctgaactg 540

gtaccagcag aaacctggca aggcccctaa gctgctgatc tacggcgcca ccaacctggc 600

cgatggcgtg cctagtagat tcagcggatc tggcagcggc acagacttca ccctgaccat 660

cagcagcctg caacctgagg actttgccac atactactgc cagaacgtgc tgaatacacc 720

tctgacattc ggccaaggaa ccaaagtgga aatcaagcgg accgtggccg ctcctagcgt 780

gttcatcttc cctccttccg atgaacaact gaagagcgga accgcctctg tggtgtgcct 840

gctgaacaac ttctacccta gagaggccaa ggtgcagtgg aaggtcgaca acgccctgca 900

gagcggcaac agccaggaga gcgtgacgga acaggacagc aaggacagca cctacagcct 960

gagctccacc cttacactgt ctaaagccga ctacgagaag cacaaggtgt acgcctgtga 1020

agtgacacac cagggcctga gcagccctgt gaccaagtct tttaaccggg gcgagtgctg 1080

aattcgaatc gtacctaggg atccagacat gataagatac attgatgagt ttggacaaac 1140

cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 1200

atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 1260

gtttcaggtt cagggggagg tgtgggaggt tttttaa 1297

<210> 94

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 94

caagtgcagc tggttcaaag cggagccgaa gtgaaaaagc ccggagctag cgtgaaggtg 60

tcctgcaagg ccagcggcta tatcttcagc aactactgga tccagtgggt gcggcaggcc 120

cctggccaag gcctggaatg gatgggcgag atcctgccag gatctggctc taccgagtac 180

acagagaact tcaaggatag agtgaccatg accagagata cctccacaag caccgtgtac 240

atggaactga gcagcctgag aagcgaggac acagctgtgt actactgcgc cagatacttt 300

tttggctcat cccctaactg gtacttcgac gtgtggggcc aaggcaccct tgtcaccgtc 360

agcagcgcca gtactaaggg acccagcgtg ttcccactgg ccccatgcag cagaagcaca 420

tctgaaagca cagccgccct gggttgtctg gtcaaagact acttccccga acccgtgaca 480

gtgagctgga acagcggcgc cctgacaagc ggcgtgcaca ccttcccagc cgtgctgcag 540

agctctggcc tgtattctct gagtagcgtg gtcaccgttc ctagctccaa cttcggcaca 600

cagacctaca cctgtaatgt ggaccacaag cctagcaaca ccaaagtgga taagaccgtt 660

gagagaaagt gctgcgtgga atgcccacca tgtccagctc caccagtcgc cggcccttct 720

gttttcctgt tcccaccaaa gcccaaagac accctgatga tcagccggac ccctgaagtg 780

acatgtgtgg tggtggatgt gtcccaggag gatcctgagg tgcagtttaa ttggtacgtt 840

gacggagtgg aagttcataa tgccaagacc aaaccccgcg aggaacagtt taacagcacc 900

taccgggtgg tgtccgtgct gacagtgctg caccaggact ggctgaacgg caaagagtac 960

aagtgcaagg tgtctaacaa gggcctgcct agcagcatcg agaagaccat ctcaaaggcc 1020

aagggccagc ccagagagcc ccaagtctat acactgcctc cttctcaaga agaaatgaca 1080

aagaaccaag tgtctctgac ctgcctggtg aagggcttct accccagcga catcgccgtc 1140

gaatgggaga gcaacggaca gcctgaaaac aactacaaga cgacccctcc agtgctggac 1200

agcgatggca gcttcttcct gtattcacgg ctgaccgtgg acaagagccg atggcaagag 1260

ggcaacgtgt ttagctgcag cgtgatgcac gaagccctgc acaaccacta cacccagaag 1320

tccctgagcc tgtctctggg aaaa 1344

<210> 95

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 95

caagtacagc ttgtacaaag tggcgctgaa gtaaaaaagc caggggcttc agtgaaagtg 60

tcctgcaagg cctcgggcta catcttctcc aactactgga tccagtgggt gcgccaggcc 120

ccgggccaag ggctggagtg gatgggagag attttacctg gcagtggctc cacagagtac 180

acagagaact tcaaggacag agtcaccatg acccgggaca ccagcacatc aactgtctac 240

atggagctga gctccctccg atcagaagac acagctgtct actactgtgc ccgctacttc 300

tttggcagca gccccaattg gtattttgat gtgtggggcc aaggcacctt ggtcaccgtc 360

tcctcagcat caacaaaagg tccttctgtc ttccccctgg ccccctgcag ccgcagcaca 420

tcagaatcca cagcagccct gggctgcctg gtaaaagact acttcccaga acctgtcact 480

gtctcctgga acagtggagc cctgacatct ggtgtccaca ccttccctgc tgtcctccag 540

agctctggcc tctacagcct ctcctcagtg gtcacagttc catcctccaa ctttggcaca 600

cagacctaca cctgcaatgt ggaccacaag ccttccaaca ccaaggtgga caagactgta 660

gagaggaagt gctgtgtgga atgcccaccc tgcccagctc cacctgttgc tgggccttct 720

gtcttcctct tccctccaaa gcccaaagac accctcatga tcagccgcac cccagaagtg 780

acctgtgtgg tggtggatgt ctcccaggaa gaccctgagg tgcagttcaa ctggtatgta 840

gatggtgttg aagttcataa tgccaagacc aagccccggg aggagcagtt caacagcacc 900

tacagagtgg tgtctgtgct gacagtgctg caccaggact ggctgaatgg aaaagaatac 960

aagtgcaaag tttccaacaa gggcctgcct tcctccattg agaagaccat ctccaaagcc 1020

aagggacagc cccgggagcc ccaagtctac acacttcctc cttctcaaga agaaatgaca 1080

aagaaccaag tgtctctgac ctgcctggtg aagggcttct acccaagtga cattgctgta 1140

gaatgggaga gcaatggaca gcctgaaaac aactacaaaa cgaccccacc tgtgctggac 1200

tctgatggca gcttcttctt gtattcacgc ctgactgtgg acaagagccg ctggcaagaa 1260

ggaaatgtat tttcctgctc tgtgatgcat gaggccctgc acaatcacta cacccagaag 1320

tcactcagcc tctctctggg aaaa 1344

<210> 96

<211> 57

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 96

atgggctgga gctgcatcat cctcttcctg gtggccacgg ccacaggtgt ccacagc 57

<210> 97

<211> 1401

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 97

atgggctgga gctgcatcat cctcttcctg gtggccacgg ccacaggtgt ccacagccaa 60

gtacagcttg tacaaagtgg cgctgaagta aaaaagccag gggcttcagt gaaagtgtcc 120

tgcaaggcct cgggctacat cttctccaac tactggatcc agtgggtgcg ccaggccccg 180

ggccaagggc tggagtggat gggagagatt ttacctggca gtggctccac agagtacaca 240

gagaacttca aggacagagt caccatgacc cgggacacca gcacatcaac tgtctacatg 300

gagctgagct ccctccgatc agaagacaca gctgtctact actgtgcccg ctacttcttt 360

ggcagcagcc ccaattggta ttttgatgtg tggggccaag gcaccttggt caccgtctcc 420

tcagcatcaa caaaaggtcc ttctgtcttc cccctggccc cctgcagccg cagcacatca 480

gaatccacag cagccctggg ctgcctggta aaagactact tcccagaacc tgtcactgtc 540

tcctggaaca gtggagccct gacatctggt gtccacacct tccctgctgt cctccagagc 600

tctggcctct acagcctctc ctcagtggtc acagttccat cctccaactt tggcacacag 660

acctacacct gcaatgtgga ccacaagcct tccaacacca aggtggacaa gactgtagag 720

aggaagtgct gtgtggaatg cccaccctgc ccagctccac ctgttgctgg gccttctgtc 780

ttcctcttcc ctccaaagcc caaagacacc ctcatgatca gccgcacccc agaagtgacc 840

tgtgtggtgg tggatgtctc ccaggaagac cctgaggtgc agttcaactg gtatgtagat 900

ggtgttgaag ttcataatgc caagaccaag ccccgggagg agcagttcaa cagcacctac 960

agagtggtgt ctgtgctgac agtgctgcac caggactggc tgaatggaaa agaatacaag 1020

tgcaaagttt ccaacaaggg cctgccttcc tccattgaga agaccatctc caaagccaag 1080

ggacagcccc gggagcccca agtctacaca cttcctcctt ctcaagaaga aatgacaaag 1140

aaccaagtgt ctctgacctg cctggtgaag ggcttctacc caagtgacat tgctgtagaa 1200

tgggagagca atggacagcc tgaaaacaac tacaaaacga ccccacctgt gctggactct 1260

gatggcagct tcttcttgta ttcacgcctg actgtggaca agagccgctg gcaagaagga 1320

aatgtatttt cctgctctgt gatgcatgag gccctgcaca atcactacac ccagaagtca 1380

ctcagcctct ctctgggaaa a 1401

<210> 98

<211> 642

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 98

gacatccaga tgacacagag cccttccagc ctctctgcct cggtggggga cagagtcacc 60

atcacctgtg gggcctcgga gaacatctat ggtgctctga actggtacca gcagaaacct 120

gggaaggccc ccaagctgct catctatggg gccaccaacc tggctgatgg tgtgccctcg 180

cgcttctctg gatctggaag tggcacagac ttcaccctca ccatctccag cctgcaacct 240

gaggactttg ccacctacta ctgccagaat gtgctgaaca cacccctcac ttttggacaa 300

gggaccaaag tagaaatcaa gaggacagtg gcggcgccca gtgtcttcat cttccctcca 360

agcgatgaac aactaaagtc tggaacagcc tcggtggtct gcctgctgaa caacttctac 420

ccaagagagg ccaaggtgca gtggaaagtt gataatgccc tgcagagtgg aaacagccag 480

gagagtgtca ctgaacaaga cagcaaggac agcacctaca gcctcagcag caccctcact 540

ttatcaaaag ctgactatga gaagcacaaa gtttatgcct gtgaagtaac tcaccagggc 600

ctcagctccc ctgtcaccaa gtccttcaac cgtggggagt gt 642

<210> 99

<211> 66

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 99

atggacatga gagttcctgc acagctgctg gggctcctgc tgctctggct ctctggtgcc 60

cgctgt 66

<210> 100

<211> 708

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 100

atggacatga gagttcctgc acagctgctg gggctcctgc tgctctggct ctctggtgcc 60

cgctgtgaca tccagatgac acagagccct tccagcctct ctgcctcggt gggggacaga 120

gtcaccatca cctgtggggc ctcggagaac atctatggtg ctctgaactg gtaccagcag 180

aaacctggga aggcccccaa gctgctcatc tatggggcca ccaacctggc tgatggtgtg 240

ccctcgcgct tctctggatc tggaagtggc acagacttca ccctcaccat ctccagcctg 300

caacctgagg actttgccac ctactactgc cagaatgtgc tgaacacacc cctcactttt 360

ggacaaggga ccaaagtaga aatcaagagg acagtggcgg cgcccagtgt cttcatcttc 420

cctccaagcg atgaacaact aaagtctgga acagcctcgg tggtctgcct gctgaacaac 480

ttctacccaa gagaggccaa ggtgcagtgg aaagttgata atgccctgca gagtggaaac 540

agccaggaga gtgtcactga acaagacagc aaggacagca cctacagcct cagcagcacc 600

ctcactttat caaaagctga ctatgagaag cacaaagttt atgcctgtga agtaactcac 660

cagggcctca gctcccctgt caccaagtcc ttcaaccgtg gggagtgt 708

<210> 101

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 101

caagtgcagc tggttcaaag tggggctgaa gtaaaaaagc ctggggcttc tgtgaaggtg 60

tcctgcaagg cctcgggcta catcttctcc aactactgga tccagtgggt ccggcaggcc 120

cctgggcaag gcctggagtg gatgggagag attcttcctg gaagtggatc cacagagtac 180

acagaaaact tcaaggacag agtcaccatg accagggaca cctccacatc cacggtgtac 240

atggaacttt ccagcctgag atctgaggac actgctgtct actactgtgc ccgctacttc 300

tttggcagca gccccaattg gtattttgat gtctggggcc aagggacctt ggtcactgtc 360

tcctcagcat ccacaaaagg accctcagtc ttccctctgg ccccctgctc ccgctccaca 420

tcagaaagca cagctgccct gggttgtctt gtaaaagatt attttccaga acctgtcact 480

gtgtcctgga acagtggggc cctgacttca ggagtccaca ccttcccagc tgtgctgcag 540

agcagtggcc tctattcttt atcatctgtg gtcactgttc cttccagcaa ctttggcaca 600

cagacctaca cctgtaatgt ggaccacaaa ccttccaaca caaaagtgga caaaactgtt 660

gaaagaaaat gctgtgtgga atgtccaccc tgccctgctc caccagttgc tgggcccagt 720

gtcttcctct tccctcccaa gcccaaggac accctcatga tcagccgcac cccagaagtg 780

acctgtgtgg tggtggatgt ttctcaagaa gatcctgagg tgcagtttaa ttggtatgtc 840

gatggtgttg aagttcataa tgccaagaca aaaccccggg aggagcagtt taacagcacc 900

tacagagttg tttctgtcct cactgtgctg caccaggact ggctgaatgg aaaagaatat 960

aaatgcaaag tgagcaacaa aggcctgccc agcagcattg agaagaccat ctcaaaggcc 1020

aagggccagc cccgggagcc ccaagtatat actcttccac ccagccaaga agaaatgaca 1080

aagaaccaag tatctctcac ctgcctggtg aaaggatttt atccttctga tattgctgta 1140

gaatgggaga gtaatggaca gccagaaaac aactacaaga cgacgccgcc ggtgctggac 1200

agtgatggca gcttcttcct ctattcacgc ctcactgtgg acaagagccg ctggcaagaa 1260

ggaaatgtct tctcctgctc tgtcatgcat gaagccctgc acaaccacta cacacagaag 1320

tccctgagcc tctccctggg gaag 1344

<210> 102

<211> 57

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 102

atgggctgga gctgcatcat cctcttcctg gtggccacag ccacaggtgt ccacagc 57

<210> 103

<211> 1401

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 103

atgggctgga gctgcatcat cctcttcctg gtggccacag ccacaggtgt ccacagccaa 60

gtgcagctgg ttcaaagtgg ggctgaagta aaaaagcctg gggcttctgt gaaggtgtcc 120

tgcaaggcct cgggctacat cttctccaac tactggatcc agtgggtccg gcaggcccct 180

gggcaaggcc tggagtggat gggagagatt cttcctggaa gtggatccac agagtacaca 240

gaaaacttca aggacagagt caccatgacc agggacacct ccacatccac ggtgtacatg 300

gaactttcca gcctgagatc tgaggacact gctgtctact actgtgcccg ctacttcttt 360

ggcagcagcc ccaattggta ttttgatgtc tggggccaag ggaccttggt cactgtctcc 420

tcagcatcca caaaaggacc ctcagtcttc cctctggccc cctgctcccg ctccacatca 480

gaaagcacag ctgccctggg ttgtcttgta aaagattatt ttccagaacc tgtcactgtg 540

tcctggaaca gtggggccct gacttcagga gtccacacct tcccagctgt gctgcagagc 600

agtggcctct attctttatc atctgtggtc actgttcctt ccagcaactt tggcacacag 660

acctacacct gtaatgtgga ccacaaacct tccaacacaa aagtggacaa aactgttgaa 720

agaaaatgct gtgtggaatg tccaccctgc cctgctccac cagttgctgg gcccagtgtc 780

ttcctcttcc ctcccaagcc caaggacacc ctcatgatca gccgcacccc agaagtgacc 840

tgtgtggtgg tggatgtttc tcaagaagat cctgaggtgc agtttaattg gtatgtcgat 900

ggtgttgaag ttcataatgc caagacaaaa ccccgggagg agcagtttaa cagcacctac 960

agagttgttt ctgtcctcac tgtgctgcac caggactggc tgaatggaaa agaatataaa 1020

tgcaaagtga gcaacaaagg cctgcccagc agcattgaga agaccatctc aaaggccaag 1080

ggccagcccc gggagcccca agtatatact cttccaccca gccaagaaga aatgacaaag 1140

aaccaagtat ctctcacctg cctggtgaaa ggattttatc cttctgatat tgctgtagaa 1200

tgggagagta atggacagcc agaaaacaac tacaagacga cgccgccggt gctggacagt 1260

gatggcagct tcttcctcta ttcacgcctc actgtggaca agagccgctg gcaagaagga 1320

aatgtcttct cctgctctgt catgcatgaa gccctgcaca accactacac acagaagtcc 1380

ctgagcctct ccctggggaa g 1401

<210> 104

<211> 642

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 104

gatatccaga tgacacagag ccccagctcc ttatcagcct cggtggggga cagggtcacc 60

atcacctgtg gggcctcgga gaacatctat ggggctctga actggtacca gcagaaacca 120

gggaaggccc ccaagctgct gatctatggg gccaccaacc tggctgatgg ggtgccttca 180

cgcttctctg gcagtggcag tggaactgac ttcaccctga ccatcagcag cttacaacca 240

gaggattttg ccacctacta ctgccagaat gtcctcaaca cacctctcac ttttggacaa 300

ggaaccaaag tggaaatcaa gagaacagtg gctgcaccca gtgtgttcat cttcccacca 360

agcgatgaac aactaaagag tggaacagca tctgtggtgt gcctgctcaa caacttctac 420

ccaagagagg ccaaggtgca gtggaaggtt gacaatgccc tgcagagtgg aaacagtcaa 480

gaatctgtca ctgaacaaga cagcaaggac agcacctaca gcctgagctc caccttaact 540

ttatcaaaag ctgactatga gaagcacaag gtctatgcct gtgaagtgac acaccagggc 600

ctctccagcc ctgtcaccaa atcttttaac agaggagaat gt 642

<210> 105

<211> 66

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 105

atggacatga gagtccctgc acagctgctg gggctcctgc tgctctggct ctctggggcc 60

cgctgt 66

<210> 106

<211> 708

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 106

atggacatga gagtccctgc acagctgctg gggctcctgc tgctctggct ctctggggcc 60

cgctgtgata tccagatgac acagagcccc agctccttat cagcctcggt gggggacagg 120

gtcaccatca cctgtggggc ctcggagaac atctatgggg ctctgaactg gtaccagcag 180

aaaccaggga aggcccccaa gctgctgatc tatggggcca ccaacctggc tgatggggtg 240

ccttcacgct tctctggcag tggcagtgga actgacttca ccctgaccat cagcagctta 300

caaccagagg attttgccac ctactactgc cagaatgtcc tcaacacacc tctcactttt 360

ggacaaggaa ccaaagtgga aatcaagaga acagtggctg cacccagtgt gttcatcttc 420

ccaccaagcg atgaacaact aaagagtgga acagcatctg tggtgtgcct gctcaacaac 480

ttctacccaa gagaggccaa ggtgcagtgg aaggttgaca atgccctgca gagtggaaac 540

agtcaagaat ctgtcactga acaagacagc aaggacagca cctacagcct gagctccacc 600

ttaactttat caaaagctga ctatgagaag cacaaggtct atgcctgtga agtgacacac 660

cagggcctct ccagccctgt caccaaatct tttaacagag gagaatgt 708

<210> 107

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 107

caagtgcaat tggtccagtc aggtgcagag gttaagaagc ctggggcatc tgttaaggtt 60

tcctgcaagg catcaggtta cattttcagc aactactgga ttcaatgggt gaggcaggca 120

ccaggtcaag gattggaatg gatgggggaa atactgcctg ggtcaggatc cacagagtac 180

acagagaact tcaaagatag ggtcaccatg actagagaca catctactag cactgtttac 240

atggagctca gtagccttag gtcagaagac actgctgtct actactgtgc cagatatttc 300

tttggcagca gccctaattg gtactttgat gtatggggcc agggcaccct ggtgactgtg 360

agcagtgctt ccacaaaggg cccatcagtc ttcccattgg caccttgtag caggagcact 420

tcagagagca cagctgcact gggttgcttg gtgaaggact acttcccaga accagtgaca 480

gtgtcctgga acagtggtgc acttacatca ggagtgcaca ccttccctgc agtactccaa 540

tcaagtggcc tttacagcct ctccagcgtt gtcacagtcc cctcatctaa ctttggaact 600

cagacctata cctgtaatgt ggaccacaag ccttccaaca ccaaagtgga caagacagta 660

gaaaggaaat gctgtgtgga gtgcccaccg tgcccagccc caccagttgc aggcccaagt 720

gtgttcctct tcccccccaa gcccaaagac accctgatga tcagtaggac ccctgaggtg 780

acctgtgttg tggtggatgt gagccaggag gatcctgagg tgcagtttaa ttggtacgtt 840

gacggagtgg aagttcataa tgccaaaact aagcctaggg aggagcagtt caatagcacc 900

tacagggtgg tgtctgttct tacagtcctg caccaagact ggctgaatgg caaagaatac 960

aagtgcaaag tcagcaacaa ggggctgcct agctctattg agaagaccat cagcaaagcc 1020

aaaggacagc ctagagaacc ccaggtgtat accttgcctc cctcccaaga agagatgacc 1080

aagaaccaag tgagcctgac ttgccttgtg aagggcttct acccttcaga tatagctgtt 1140

gagtgggaga gcaatggcca gccagaaaac aactacaaaa ccaccccacc tgtattggat 1200

agtgatggaa gctttttctt gtacagcagg ctgactgttg ataagagcag gtggcaggag 1260

ggcaatgtgt tcagttgttc tgtgatgcat gaggccctgc acaatcacta cacccagaag 1320

agtctgtccc tttctctggg caag 1344

<210> 108

<211> 57

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 108

atgggctggt catgtatcat cctgttcctg gtagccactg ccacaggggt tcatagc 57

<210> 109

<211> 1401

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 109

atgggctggt catgtatcat cctgttcctg gtagccactg ccacaggggt tcatagccaa 60

gtgcaattgg tccagtcagg tgcagaggtt aagaagcctg gggcatctgt taaggtttcc 120

tgcaaggcat caggttacat tttcagcaac tactggattc aatgggtgag gcaggcacca 180

ggtcaaggat tggaatggat gggggaaata ctgcctgggt caggatccac agagtacaca 240

gagaacttca aagatagggt caccatgact agagacacat ctactagcac tgtttacatg 300

gagctcagta gccttaggtc agaagacact gctgtctact actgtgccag atatttcttt 360

ggcagcagcc ctaattggta ctttgatgta tggggccagg gcaccctggt gactgtgagc 420

agtgcttcca caaagggccc atcagtcttc ccattggcac cttgtagcag gagcacttca 480

gagagcacag ctgcactggg ttgcttggtg aaggactact tcccagaacc agtgacagtg 540

tcctggaaca gtggtgcact tacatcagga gtgcacacct tccctgcagt actccaatca 600

agtggccttt acagcctctc cagcgttgtc acagtcccct catctaactt tggaactcag 660

acctatacct gtaatgtgga ccacaagcct tccaacacca aagtggacaa gacagtagaa 720

aggaaatgct gtgtggagtg cccaccgtgc ccagccccac cagttgcagg cccaagtgtg 780

ttcctcttcc cccccaagcc caaagacacc ctgatgatca gtaggacccc tgaggtgacc 840

tgtgttgtgg tggatgtgag ccaggaggat cctgaggtgc agtttaattg gtacgttgac 900

ggagtggaag ttcataatgc caaaactaag cctagggagg agcagttcaa tagcacctac 960

agggtggtgt ctgttcttac agtcctgcac caagactggc tgaatggcaa agaatacaag 1020

tgcaaagtca gcaacaaggg gctgcctagc tctattgaga agaccatcag caaagccaaa 1080

ggacagccta gagaacccca ggtgtatacc ttgcctccct cccaagaaga gatgaccaag 1140

aaccaagtga gcctgacttg ccttgtgaag ggcttctacc cttcagatat agctgttgag 1200

tgggagagca atggccagcc agaaaacaac tacaaaacca ccccacctgt attggatagt 1260

gatggaagct ttttcttgta cagcaggctg actgttgata agagcaggtg gcaggagggc 1320

aatgtgttca gttgttctgt gatgcatgag gccctgcaca atcactacac ccagaagagt 1380

ctgtcccttt ctctgggcaa g 1401

<210> 110

<211> 642

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 110

gatatccaga tgacccagag ccccagctcc ctgtctgcat ctgtaggtga cagggtcacc 60

attacctgtg gagcatcaga aaacatctat ggggccttga actggtatca gcagaagcca 120

ggcaaagccc caaagctgtt gatatatggt gccaccaact tggcagatgg tgtgccaagc 180

agattcagtg gatcaggcag tggcacagat ttcacactga ccattagcag cctgcaacct 240

gaagactttg ctacctacta ctgccagaat gttctgaaca cccccctgac ctttggccag 300

ggcaccaagg tggagatcaa gaggactgtt gctgcccctt ctgtattcat cttcccaccc 360

agtgatgagc aattgaagtc aggcactgca tcagtggtgt gtcttcttaa caacttctac 420

cccagagagg ccaaggtaca atggaaggtt gacaatgcac ttcagagtgg aaacagccag 480

gagtcagtca ctgaacagga cagcaaggat agcacataca gcctgtctag caccctgact 540

ctgagcaagg ctgactatga gaagcataag gtgtatgcct gtgaggttac ccaccaggga 600

ctgagcagcc ctgtgacaaa aagcttcaat aggggggagt gc 642

<210> 111

<211> 66

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 111

atggacatga gggtgccagc acagctgctg ggcctcctgc tgctgtggct gagtggtgca 60

agatgt 66

<210> 112

<211> 708

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 112

atggacatga gggtgccagc acagctgctg ggcctcctgc tgctgtggct gagtggtgca 60

agatgtgata tccagatgac ccagagcccc agctccctgt ctgcatctgt aggtgacagg 120

gtcaccatta cctgtggagc atcagaaaac atctatgggg ccttgaactg gtatcagcag 180

aagccaggca aagccccaaa gctgttgata tatggtgcca ccaacttggc agatggtgtg 240

ccaagcagat tcagtggatc aggcagtggc acagatttca cactgaccat tagcagcctg 300

caacctgaag actttgctac ctactactgc cagaatgttc tgaacacccc cctgaccttt 360

ggccagggca ccaaggtgga gatcaagagg actgttgctg ccccttctgt attcatcttc 420

ccacccagtg atgagcaatt gaagtcaggc actgcatcag tggtgtgtct tcttaacaac 480

ttctacccca gagaggccaa ggtacaatgg aaggttgaca atgcacttca gagtggaaac 540

agccaggagt cagtcactga acaggacagc aaggatagca catacagcct gtctagcacc 600

ctgactctga gcaaggctga ctatgagaag cataaggtgt atgcctgtga ggttacccac 660

cagggactga gcagccctgt gacaaaaagc ttcaataggg gggagtgc 708

<210> 113

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 113

caagtacagc ttgtacaaag tggcgctgaa gtaaaaaagc caggggcttc agtgaaagtg 60

tcctgcaagg cctcgggcca catcttctcc aactactgga tccagtgggt gcgccaggcc 120

ccgggccaag ggctggagtg gatgggagag attttacctg gcagtggcca cacagagtac 180

acagagaact tcaaggacag agtcaccatg acccgggaca ccagcacatc aactgtctac 240

atggagctga gctccctccg atcagaagac acagctgtct actactgtgc ccgctacttc 300

tttggcagca gccccaattg gtattttgat gtgtggggcc aaggcacctt ggtcaccgtc 360

tcctcagcat caacaaaagg tccttctgtc ttccccctgg ccccctgcag ccgcagcaca 420

tcagaatcca cagcagccct gggctgcctg gtaaaagact acttcccaga acctgtcact 480

gtctcctgga acagtggagc cctgacatct ggtgtccaca ccttccctgc tgtcctccag 540

agctctggcc tctacagcct ctcctcagtg gtcacagttc catcctccaa ctttggcaca 600

cagacctaca cctgcaatgt ggaccacaag ccttccaaca ccaaggtgga caagactgta 660

gagaggaagt gctgtgtgga atgcccaccc tgcccagctc cacctgttgc tgggccttct 720

gtcttcctct tccctccaaa gcccaaagac accctcatga tcagccgcac cccagaagtg 780

acctgtgtgg tggtggatgt ctcccaggaa gaccctgagg tgcagttcaa ctggtatgta 840

gatggtgttg aagttcataa tgccaagacc aagccccggg aggagcagtt caacagcacc 900

tacagagtgg tgtctgtgct gacagtgctg caccaggact ggctgaatgg aaaagaatac 960

aagtgcaaag tttccaacaa gggcctgcct tcctccattg agaagaccat ctccaaagcc 1020

aagggacagc cccgggagcc ccaagtctac acacttcctc cttctcaaga agaaatgaca 1080

aagaaccaag tgtctctgac ctgcctggtg aagggcttct acccaagtga cattgctgta 1140

gaatgggaga gcaatggaca gcctgaaaac aactacaaaa cgaccccacc tgtgctggac 1200

tctgatggca gcttcttctt gtattcacgc ctgactgtgg acaagagccg ctggcaagaa 1260

ggaaatgtat tttcctgctc tgtgctccat gaggccctgc acagccacta cacccagaag 1320

tcactcagcc tctctctggg aaaa 1344

<210> 114

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 114

caagtgcagc tggttcaaag tggggctgaa gtaaaaaagc ctggggcttc tgtgaaggtg 60

tcctgcaagg cctcgggcca catcttctcc aactactgga tccagtgggt ccggcaggcc 120

cctgggcaag gcctggagtg gatgggagag attcttcctg gaagtggaca cacagagtac 180

acagaaaact tcaaggacag agtcaccatg accagggaca cctccacatc cacggtgtac 240

atggaacttt ccagcctgag atctgaggac actgctgtct actactgtgc ccgctacttc 300

tttggcagca gccccaattg gtattttgat gtctggggcc aagggacctt ggtcactgtc 360

tcctcagcat ccacaaaagg accctcagtc ttccctctgg ccccctgctc ccgctccaca 420

tcagaaagca cagctgccct gggttgtctt gtaaaagatt attttccaga acctgtcact 480

gtgtcctgga acagtggggc cctgacttca ggagtccaca ccttcccagc tgtgctgcag 540

agcagtggcc tctattcttt atcatctgtg gtcactgttc cttccagcaa ctttggcaca 600

cagacctaca cctgtaatgt ggaccacaaa ccttccaaca caaaagtgga caaaactgtt 660

gaaagaaaat gctgtgtgga atgtccaccc tgccctgctc caccagttgc tgggcccagt 720

gtcttcctct tccctcccaa gcccaaggac accctcatga tcagccgcac cccagaagtg 780

acctgtgtgg tggtggatgt ttctcaagaa gatcctgagg tgcagtttaa ttggtatgtc 840

gatggtgttg aagttcataa tgccaagaca aaaccccggg aggagcagtt taacagcacc 900

tacagagttg tttctgtcct cactgtgctg caccaggact ggctgaatgg aaaagaatat 960

aaatgcaaag tgagcaacaa aggcctgccc agcagcattg agaagaccat ctcaaaggcc 1020

aagggccagc cccgggagcc ccaagtatat actcttccac ccagccaaga agaaatgaca 1080

aagaaccaag tatctctcac ctgcctggtg aaaggatttt atccttctga tattgctgta 1140

gaatgggaga gtaatggaca gccagaaaac aactacaaga cgacgccgcc ggtgctggac 1200

agtgatggca gcttcttcct ctattcacgc ctcactgtgg acaagagccg ctggcaagaa 1260

ggaaatgtct tctcctgctc tgtcctccat gaagccctgc acagccacta cacacagaag 1320

tccctgagcc tctccctggg gaag 1344

<210> 115

<211> 1344

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 115

caagtgcaat tggtccagtc aggtgcagag gttaagaagc ctggggcatc tgttaaggtt 60

tcctgcaagg catcaggtca cattttcagc aactactgga ttcaatgggt gaggcaggca 120

ccaggtcaag gattggaatg gatgggggaa atactgcctg ggtcaggaca cacagagtac 180

acagagaact tcaaagatag ggtcaccatg actagagaca catctactag cactgtttac 240

atggagctca gtagccttag gtcagaagac actgctgtct actactgtgc cagatatttc 300

tttggcagca gccctaattg gtactttgat gtatggggcc agggcaccct ggtgactgtg 360

agcagtgctt ccacaaaggg cccatcagtc ttcccattgg caccttgtag caggagcact 420

tcagagagca cagctgcact gggttgcttg gtgaaggact acttcccaga accagtgaca 480

gtgtcctgga acagtggtgc acttacatca ggagtgcaca ccttccctgc agtactccaa 540

tcaagtggcc tttacagcct ctccagcgtt gtcacagtcc cctcatctaa ctttggaact 600

cagacctata cctgtaatgt ggaccacaag ccttccaaca ccaaagtgga caagacagta 660

gaaaggaaat gctgtgtgga gtgcccaccg tgcccagccc caccagttgc aggcccaagt 720

gtgttcctct tcccccccaa gcccaaagac accctgatga tcagtaggac ccctgaggtg 780

acctgtgttg tggtggatgt gagccaggag gatcctgagg tgcagtttaa ttggtacgtt 840

gacggagtgg aagttcataa tgccaaaact aagcctaggg aggagcagtt caatagcacc 900

tacagggtgg tgtctgttct tacagtcctg caccaagact ggctgaatgg caaagaatac 960

aagtgcaaag tcagcaacaa ggggctgcct agctctattg agaagaccat cagcaaagcc 1020

aaaggacagc ctagagaacc ccaggtgtat accttgcctc cctcccaaga agagatgacc 1080

aagaaccaag tgagcctgac ttgccttgtg aagggcttct acccttcaga tatagctgtt 1140

gagtgggaga gcaatggcca gccagaaaac aactacaaaa ccaccccacc tgtattggat 1200

agtgatggaa gctttttctt gtacagcagg ctgactgttg ataagagcag gtggcaggag 1260

ggcaatgtgt tcagttgttc tgtgctccat gaggccctgc acagccacta cacccagaag 1320

agtctgtccc tttctctggg caag 1344

<210> 116

<211> 410

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 116

tgcatgtata atttctacag aacctattag aaaggatcac ccagcctctg cttttgtaca 60

actttccctt aaaaaactgc caattccact gctgtttggc ccaatagtga gaactttttc 120

ctgctgcctc ttggtgcttt tgcctatggc ccctattctg cctgctgaag acactcttgc 180

cagcatggac ttaaacccct ccagctctga caatcctctt tctcttttgt tttacatgaa 240

gggtctggca gccaaagcaa tcactcaaag ttcaaacctt atcatttttt gctttgttcc 300

tcttggcctt ggttttgtac atcagctttg aaaataccat cccagggtta atgctggggt 360

taatttataa ctaagagtgc tctagttttg caatacagga catgctataa 410

<210> 117

<211> 222

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 117

ctttctcttt tgttttacat gaagggtctg gcagccaaag caatcactca aagttcaaac 60

cttatcattt tttgctttgt tcctcttggc cttggttttg tacatcagct ttgaaaatac 120

catcccaggg ttaatgctgg ggttaattta taactaagag tgctctagtt ttgcaataca 180

ggacatgcta taaaaatgga aagatgttgc tttctgagag at 222

<210> 118

<211> 402

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 118

accagtggaa cagccactaa ggattctgca gtgagagcag agggccagct aagtggtact 60

ctcccagaga ctgtctgact cacgccaccc cctccacctt ggacacagga cgctgtggtt 120

tctgagccag gtacaatgac tcctttcggt aagtgcagtg gaagctgtac actgcccagg 180

caaagcgtcc gggcagcgta ggcgggcgac tcagatccca gccagtggac ttagcccctg 240

tttgctcctc cgataactgg ggtgaccttg gttaatattc accagcagcc tcccccgttg 300

cccctctgga tccactgctt aaatacggac gaggacaggg ccctgtctcc tcagcttcag 360

gcaccaccac tgacctggga cagtgaatcg taagtatgcc tt 402

<210> 119

<211> 202

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 119

acctattaag aatatttcat agaacgaatg ttccgatgct ctaatctctc tagacaaggt 60

tcatatttgt atgggttact tattctctct ttgttgacta agtcaataat cagaatcagc 120

aggtttgcag tcagattggc agggataagc agcctagctc aggagaagtg agtataaaag 180

ccccaggctg ggagcagcca tc 202

<210> 120

<211> 392

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 120

ttctgatatc tatttaactg atttcaccca aatgctttga acctgggaat gtacctctcc 60

ccctccccca cccccaacag gagtgagaca agggccaggg ctattgcccc tgctgactca 120

atattggcta atcactgcct agaactgata aggtgatcaa atgaccaggt gccttcaacc 180

tttaccctgg tagaagcctc ttattcacct cttttcctgc cagagccctc cattgggagg 240

ggacgggcgg aagctgtttt ctgaatttgt tttactgggg gtagggtatg ttcagtgatc 300

gtccctgtca cctgacaggg ggtgggtaaa cagacaggta tatagcccct tcctctccag 360

ccagggcagg cacagacacc aaggacagag ac 392

<210> 121

<211> 210

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 121

agttccagat ggtaaatata cacaagggat ttagtcaaac aattttttgg caagaatatt 60

atgaattttg taatcggttg gcagccaatg aaatacaaag atgagtctag ttaataatct 120

acaattattg gttaaagaag tatattagtg ctaatttccc tccgtttgtc ctagcttttc 180

tcttctgtca accccacacg cctttggcac 210

<210> 122

<211> 113

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 122

tgttgcttaa atgtttgttg actaagtcaa taatcagaat cagcaaatta aatatttaac 60

taaggaaact aggcaaggtt catatttatt cctagcagag gactcagata taa 113

<210> 123

<211> 325

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 123

ctgcaggctc agaggcacac aggagtttct gggctcaccc tgcccccttc caacccctca 60

gttcccatcc tccagcagct gtttgtgtgc tgcctctgaa gtccacactg aacaaacttc 120

agcctactca tgtccctaaa atgggcaaac attgcaagca gcaaacagca aacacacagc 180

cctccctgcc tgctgacctt ggagctgggg cagaggtcag agacctctct gggcccatgc 240

cacctccaac atccactcga ccccttggaa tttcggtgga gaggagcaga ggttgtcctg 300

gcgtggttta ggtagtgtga gaggg 325

<210> 124

<211> 299

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 124

gtttttgggc ccgccctgcc cccttccgac ctcttagttc ctatcctcca gcagctgttt 60

gtgtgctgcc tctgaagtcc accctgaatg accttcagcc tgttcccgtc cctgatatgg 120

gcaaacattg caagcagcaa acagcaaaca catagccctc cctgcgtgct gaccttggag 180

ctgcggcaga ggtcagagac ctctcagggc ccataccact tccaacatcc ccttgatctc 240

ttggattttg gtggagaggg gcagaggttg tcctggcctg gttaggtagt gtgagaggg 299

<210> 125

<211> 54

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 125

gaaggcagag gcagcctgct gacctgtgga gatgtggaag agaacccagg ccct 54

<210> 126

<211> 9

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 126

ggctctgga 9

<210> 127

<211> 75

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 127

agaaagagaa gaggctctgg agaaggcaga ggcagcctgc tgacctgtgg agatgtggaa 60

gagaacccag gccct 75

<210> 128

<211> 57

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 128

gctactaact tcagcctgct gaagcaggct ggagatgtgg aggagaaccc tggacct 57

<210> 129

<211> 78

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 129

agaaagagaa gaggctctgg agctactaac ttcagcctgc tgaagcaggc tggagatgtg 60

gaggagaacc ctggacct 78

<210> 130

<211> 66

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 130

atggacatga gggtgccagc acagctgctg ggcctcctgc tgctgtggct gagtggtgca 60

aggtgt 66

<210> 131

<211> 642

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 131

gatatccaga tgacccagag ccccagctcc ctgtctgcat ctgtaggtga cagggtcacc 60

attacctgtg gagcatcaga aaacatctat ggggccttga actggtatca gcagaagcca 120

ggtaaagccc caaagctgtt gatatatggt gccaccaact tggcagatgg tgtgccaagc 180

agattcagtg gatcaggcag tggcacagat ttcacactga ccattagcag cctgcaacct 240

gaagactttg ctacctacta ctgccagaat gttctgaaca cccccctgac ctttggccag 300

ggcaccaagg tggagatcaa gaggactgtt gctgcccctt ctgtattcat cttcccaccc 360

agtgatgagc aattgaagtc aggcactgca tcagtggtgt gtcttcttaa caacttctac 420

cccagagagg ccaaggtaca atggaaggtt gacaatgcac ttcagagtgg aaacagccag 480

gagtcagtca ctgaacagga cagcaaggat agcacataca gcctgtctag caccctgact 540

ctgagcaagg ctgactatga gaagcataag gtgtatgcct gtgaggttac ccaccaggga 600

ctgagcagcc ctgtgacaaa aagcttcaat aggggggagt gc 642

<210> 132

<211> 708

<212> DNA

<213> artificial sequence

<220>

<223> synthetic nucleic acid

<400> 132

atggacatga gggtgccagc acagctgctg ggcctcctgc tgctgtggct gagtggtgca 60

aggtgtgata tccagatgac ccagagcccc agctccctgt ctgcatctgt aggtgacagg 120

gtcaccatta cctgtggagc atcagaaaac atctatgggg ccttgaactg gtatcagcag 180

aagccaggta aagccccaaa gctgttgata tatggtgcca ccaacttggc agatggtgtg 240

ccaagcagat tcagtggatc aggcagtggc acagatttca cactgaccat tagcagcctg 300

caacctgaag actttgctac ctactactgc cagaatgttc tgaacacccc cctgaccttt 360

ggccagggca ccaaggtgga gatcaagagg actgttgctg ccccttctgt attcatcttc 420

ccacccagtg atgagcaatt gaagtcaggc actgcatcag tggtgtgtct tcttaacaac 480

ttctacccca gagaggccaa ggtacaatgg aaggttgaca atgcacttca gagtggaaac 540

agccaggagt cagtcactga acaggacagc aaggatagca catacagcct gtctagcacc 600

ctgactctga gcaaggctga ctatgagaag cataaggtgt atgcctgtga ggttacccac 660

cagggactga gcagccctgt gacaaaaagc ttcaataggg gggagtgc 708

Claims

1. A recombinant adeno-associated virus (rAAV) genome, the rAAV genome comprising:

(a) A first expression cassette comprising, from 5 'to 3':

a first liver-specific transcription regulatory element,

A first polyadenylation sequence; and

(b) A second expression cassette comprising, from 5 'to 3':

a second liver-specific transcription regulatory element,

2. The rAAV genome of claim 1, wherein the first transcriptional regulatory element and/or the second transcriptional regulatory element comprises a promoter element selected from the group consisting of: human albumin promoter, human transthyretin (TTR) promoter, human thyroxine-binding globulin (TBG) promoter, human ApoH promoter, human SERPINA1 (hAAT) promoter and liver-specific regulatory modules thereof, such as human ApoE/C-I liver control region (HCR) 1 or 2.

3. The rAAV genome of claim 1 or 2, wherein the first transcription regulatory element and/or the second transcription regulatory element comprises a promoter element comprising a nucleic acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs 25, 27, 66, 68, 69, 116 and 117.

4. A rAAV genome according to any one of claims 1 to 3, wherein the transcriptional regulatory element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 27.

5. The rAAV genome according to any one of claims 1 to 4, wherein the transcriptional regulatory element comprises the nucleotide sequence set forth in SEQ ID No. 27.

6. The rAAV genome according to any one of claims 1 to 5, wherein the nucleotide sequence of the transcriptional regulatory element consists of the nucleotide sequence set forth in SEQ ID No. 27.

7. A rAAV genome according to any one of claims 1 to 3, wherein the transcriptional regulatory element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 67.

8. The rAAV genome of any one of claims 1-4, wherein the transcriptional regulatory element comprises the nucleotide sequence set forth in SEQ ID No. 67.

9. The rAAV genome according to any one of claims 1 to 5, wherein the nucleotide sequence of the transcriptional regulatory element consists of the nucleotide sequence set forth in SEQ ID No. 67.

10. The rAAV genome of any one of claims 1-9, wherein the first expression cassette and/or the second expression cassette further comprises an intron element that is located 5 'relative to the first coding sequence and/or the second coding sequence and 3' relative to the transcriptional regulatory element.

11. The rAAV genome of claim 10, wherein the intronic element is an exogenous intronic element, optionally wherein the exogenous intronic element is an SV40 intronic element or a mouse picovirus (MVM) intronic element.

12. The rAAV genome of claim 11, wherein the SV40 intron element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence shown in SEQ ID No. 29.

13. The rAAV genome of claim 11 or 12, wherein the SV40 intron element comprises the nucleotide sequence shown in SEQ ID No. 29.

14. The rAAV genome according to any one of claims 11 to 13, wherein the nucleotide sequence of the SV40 intron element consists of the nucleotide sequence shown in SEQ ID No. 29.

15. The rAAV genome of any one of claims 11-14, wherein the MVM intron element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 30.

16. The rAAV genome of any one of claims 11-14, wherein the MVM intron element comprises the nucleotide sequence set forth in SEQ ID No. 30.

17. The rAAV genome of any one of claims 11-16, wherein the nucleotide sequence of the MVM intron element consists of the nucleotide sequence set forth in SEQ ID No. 30.

18. The rAAV genome of any one of claims 1-17, wherein the first transcriptional regulatory element and the second transcriptional regulatory element are the same.

19. The rAAV genome of any one of claims 1-18, wherein the first transcriptional regulatory element comprises an HCR 1 element, an hAAT promoter, and an SV40 intron element, and the second transcriptional regulatory element comprises a SERPINA1 liver-specific regulatory module, a TTR promoter, and an MVM intron element.

20. The rAAV genome of any one of claims 1-19, wherein the first transcriptional regulatory element comprises a nucleic acid sequence of SEQ ID No. 50 and the second transcriptional regulatory element comprises a nucleic acid sequence of SEQ ID No. 43.

21. The rAAV genome of any one of claims 1-20, wherein the first expression cassette and/or the second expression cassette further comprises a polyadenylation sequence 3' relative to the first coding sequence and/or the second coding sequence.

22. The rAAV genome of claim 21, wherein the polyadenylation sequence is an exogenous polyadenylation sequence, optionally wherein the exogenous polyadenylation sequence is an SV40 polyadenylation sequence or a Bovine Growth Hormone (BGH) polyadenylation sequence.

23. The rAAV genome of claim 22, wherein the SV40 polyadenylation sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence shown in SEQ ID No. 31.

24. The rAAV genome of claim 22 or 23, wherein the SV40 polyadenylation sequence comprises the nucleotide sequence shown in SEQ ID No. 31.

25. The rAAV genome of any one of claims 22-24, wherein the nucleotide sequence of the SV40 polyadenylation sequence consists of the nucleotide sequence shown in SEQ ID No. 31.

26. The rAAV genome of any one of claims 22-25, wherein the BGH polyadenylation sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 33.

27. The rAAV genome of any one of claims 22-26, wherein the BGH polyadenylation sequence comprises the nucleotide sequence set forth in SEQ ID No. 33.

28. The rAAV genome of any one of claims 22-27, wherein the nucleotide sequence of the BGH polyadenylation sequence consists of the nucleotide sequence set forth in SEQ ID No. 33.

29. The rAAV genome of any one of claims 21-28, wherein the first expression cassette and the second expression cassette comprise the same polyadenylation sequence.

30. The rAAV genome of any one of claims 21-29, wherein the first expression cassette comprises the SV40 polyadenylation sequence.

31. The rAAV genome of any one of claims 21-30, wherein the second expression cassette comprises the BGH polyadenylation sequence.

32. The rAAV genome of any one of claims 21-31, wherein the first polyadenylation sequence comprises the nucleic acid sequence of SEQ ID No. 31 and the second polyadenylation sequence comprises the nucleic acid sequence of SEQ ID No. 33.

33. The rAAV genome of any one of claims 1-32, wherein the first expression cassette and the second expression cassette are in the same orientation in the rAAV genome.

34. The rAAV genome of any one of claims 1-32, wherein the first expression cassette and the second expression cassette are in opposite orientations in the rAAV genome.

35. The rAAV genome of any one of the preceding claims, wherein the first expression cassette and the second expression cassette are in opposite orientations, the first polyadenylation sequence and the second polyadenylation sequence being distally located in the rAAV genome.

36. The rAAV genome of claim 35, wherein the rAAV genome further comprises a stuffer sequence interposed between the first transcriptional regulatory element and the second transcriptional regulatory element.

37. The rAAV genome of claim 36, wherein the stuffer sequence comprises a β -globin polyadenylation sequence.

38. The rAAV genome of claim 37, wherein the beta globin polyadenylation sequence comprises the nucleic acid sequence of SEQ ID No. 51.

39. The rAAV genome of claim 35, wherein the rAAV genome comprises, from 5 'to 3':

(a) The first polyadenylation sequence comprising the nucleic acid sequence of SEQ ID NO. 33;

(b) The first coding sequence;

(c) The first liver-specific transcription regulatory element comprising the nucleic acid sequence of SEQ ID NO. 27;

(d) A stuffer sequence comprising the nucleic acid sequence of SEQ ID NO. 51;

(e) Said second liver-specific transcriptional regulatory element comprising the nucleic acid sequence of SEQ ID NO. 67;

(f) The second coding sequence; and

(g) The second transcribed polyadenylation sequence comprising the nucleic acid sequence of SEQ ID NO. 31.

40. The rAAV genome of claim 35, wherein the rAAV genome comprises, from 5 'to 3': a reverse complement of the first expression cassette; a filling sequence; and the second expression cassette.

41. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

a nucleotide sequence which is at least 90% identical to the nucleotide sequence shown in SEQ ID NO. 27,

the first code sequence is used to encode a first code sequence,

a nucleotide sequence at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 33;

(b) The stuffer sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 51 or an inverse complement thereof; and

(c) The second expression cassette comprises, from 5 'to 3':

a nucleotide sequence which is at least 90% identical to the nucleotide sequence shown in SEQ ID NO. 67,

The second code sequence is used to encode a second code sequence,

a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 31.

42. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

the first code sequence is used to encode a first code sequence,

a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 31;

(c) The second expression cassette comprises, from 5 'to 3':

the second code sequence is used to encode a second code sequence,

a nucleotide sequence which is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO. 33.

43. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

a nucleotide sequence which is at least 90% identical to the nucleotide sequence shown in SEQ ID NO. 25,

a nucleotide sequence which is at least 90% identical to the nucleotide sequence shown in SEQ ID NO. 26,

The first code sequence is used to encode a first code sequence,

the first polyadenylation sequence;

(c) The second expression cassette comprises, from 5 'to 3':

a nucleotide sequence which is at least 90% identical to the nucleotide sequence shown in SEQ ID NO. 119,

a nucleotide sequence which is at least 90% identical to the nucleotide sequence shown in SEQ ID NO. 45,

the second code sequence is used to encode a second code sequence,

44. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

the first code sequence is used to encode a first code sequence,

(c) The second expression cassette comprises, from 5 'to 3':

the second code sequence is used to encode a second code sequence,

the first polyadenylation sequence.

45. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

the nucleotide sequence shown in SEQ ID NO. 27,

the first code sequence is used to encode a first code sequence,

the nucleotide sequence shown in SEQ ID NO. 33;

(b) The stuffer sequence comprises the nucleotide sequence set forth in SEQ ID NO. 51 or an inverse complement thereof; and

(c) The second expression cassette comprises, from 5 'to 3':

the nucleotide sequence shown in SEQ ID NO. 67,

the second code sequence is used to encode a second code sequence,

the nucleotide sequence shown in SEQ ID NO. 31.

46. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

the nucleotide sequence shown in SEQ ID NO. 67,

the first code sequence is used to encode a first code sequence,

the nucleotide sequence shown in SEQ ID NO. 31;

(c) The second expression cassette comprises, from 5 'to 3':

The nucleotide sequence shown in SEQ ID NO. 27,

the second code sequence is used to encode a second code sequence,

the nucleotide sequence shown in SEQ ID NO. 33.

47. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

the nucleotide sequence shown in SEQ ID NO. 25,

the nucleotide sequence shown in SEQ ID NO. 26,

the first code sequence is used to encode a first code sequence,

the first polyadenylation sequence;

(c) The second expression cassette comprises, from 5 'to 3':

the nucleotide sequence shown in SEQ ID NO. 119,

the nucleotide sequence shown in SEQ ID NO. 45,

the second code sequence is used to encode a second code sequence,

the nucleotide sequence shown in SEQ ID NO. 31.

48. The rAAV genome according to claim 40, wherein:

(a) The first expression cassette comprises, from 5 'to 3':

the nucleotide sequence shown in SEQ ID NO. 119,

the nucleotide sequence shown in SEQ ID NO. 45,

the first code sequence is used to encode a first code sequence,

the nucleotide sequence shown in SEQ ID NO. 31;

(c) The second expression cassette comprises, from 5 'to 3':

The nucleotide sequence shown in SEQ ID NO. 25,

the nucleotide sequence shown in SEQ ID NO. 26,

the second code sequence is used to encode a second code sequence,

the first polyadenylation sequence.

49. A rAAV genome comprising a bicistronic expression cassette comprising, from 5 'to 3':

(a) A liver-specific transcriptional regulatory element; a first coding sequence encoding a first polypeptide comprising an antibody heavy chain operably linked to a first signal sequence; a ribosome-hopping sequence encoding a ribosome-hopping peptide; a second coding sequence encoding a second polypeptide comprising an antibody light chain operably linked to a second signal sequence; a polyadenylation sequence; or (b)

50. The rAAV genome of claim 49, wherein the transcriptional regulatory element comprises a promoter element selected from the group consisting of: human albumin promoter, human transthyretin (TTR) promoter, human thyroxine-binding globulin (TBG) promoter, human ApoH promoter, human SERPINA1 (hAAT) promoter and liver-specific regulatory modules thereof, such as human ApoE/C-I liver control region (HCR) 1 or 2.

51. The rAAV genome of claim 49 or 50, wherein the transcriptional regulatory element comprises a promoter element comprising a nucleic acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs 25, 27, 66, 68, 69, 116 and 117.

52. The rAAV genome of any one of claims 49-51, wherein the transcriptional regulatory element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 27.

53. The rAAV genome of any one of claims 49-52, wherein the transcriptional regulatory element comprises the nucleotide sequence set forth in SEQ ID No. 27.

54. The rAAV genome according to any one of claims 49 to 53, wherein the nucleotide sequence of the transcriptional regulatory element consists of the nucleotide sequence set forth in SEQ ID No. 27.

55. The rAAV genome of claim 49 or 50, wherein the transcriptional regulatory element comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 67.

56. The rAAV genome of claim 55, wherein the transcriptional regulatory element comprises the nucleotide sequence set forth in SEQ ID No. 67.

57. The rAAV genome according to claim 55, wherein the nucleotide sequence of the transcriptional regulatory element consists of the nucleotide sequence set forth in SEQ ID No. 67.

58. The rAAV genome of any one of claims 49-57, wherein the bicistronic expression cassette further comprises an intron element 5 'to the first coding sequence and/or the second coding sequence and 3' to the transcriptional regulatory element.

59. The rAAV genome of claim 58, wherein the intronic element is an exogenous intronic element, optionally wherein the exogenous intronic element is an SV40 intronic element or a mouse picornavirus (MVM) intronic element.

60. The rAAV genome of claim 59, wherein the SV40 intron elements comprise a nucleotide sequence that is at least 90% identical to the nucleotide sequence depicted in SEQ ID NO. 29.

61. The rAAV genome of claim 59 or 60, wherein the SV40 intron element comprises the nucleotide sequence shown in SEQ ID No. 29.

62. The rAAV genome of any one of claims 59 to 61, wherein the nucleotide sequence of the SV40 intron element consists of the nucleotide sequence shown in SEQ ID No. 29.

63. The rAAV genome of claim 59, wherein the MVM intron element comprises a nucleotide sequence at least 90% identical to the nucleotide sequence depicted in SEQ ID NO. 30.

64. The rAAV genome of claim 59, wherein the MVM intron element comprises the nucleotide sequence depicted in SEQ ID NO. 30.

65. The rAAV genome of claim 59, wherein the nucleotide sequence of the MVM intron element consists of the nucleotide sequence depicted in SEQ ID NO. 30.

66. The rAAV genome of any one of claims 49-65, wherein the transcriptional regulatory element comprises:

(a) HCR 1 element, hAAT promoter and SV40 intron element; or (b)

(b) SERPINA1 liver-specific regulatory modules, TTR promoter, and MVM intron elements.

67. The rAAV genome of any one of claims 49-66, wherein the transcriptional regulatory element comprises a nucleic acid sequence of SEQ ID No. 50 or a nucleic acid sequence of SEQ ID No. 43.

68. The rAAV genome of any one of claims 49-67, wherein the polyadenylation sequence is an exogenous polyadenylation sequence, optionally wherein the exogenous polyadenylation sequence is an SV40 polyadenylation sequence or a Bovine Growth Hormone (BGH) polyadenylation sequence.

69. The rAAV genome of claim 68, wherein the SV40 polyadenylation sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence shown in SEQ ID No. 31.

70. The rAAV genome of claim 68 or 69, wherein the SV40 polyadenylation sequence comprises the nucleotide sequence shown in SEQ ID No. 31.

71. The rAAV genome according to any one of claims 68 to 70, wherein the nucleotide sequence of the SV40 polyadenylation sequence consists of the nucleotide sequence shown in SEQ ID No. 31.

72. The rAAV genome of any one of claims 68-71, wherein the BGH polyadenylation sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID No. 33.

73. The rAAV genome of any one of claims 68-72, wherein the BGH polyadenylation sequence comprises the nucleotide sequence set forth in SEQ ID No. 33.

74. The rAAV genome according to any of claims 68-73, wherein the nucleotide sequence of the BGH polyadenylation sequence consists of the nucleotide sequence set forth in SEQ ID No. 33.

75. The rAAV genome of any one of claims 1-74, wherein the first signal sequence and/or the second signal sequence is a naturally occurring signal sequence.

76. The rAAV genome of any one of claims 1-74, wherein the first signal sequence and/or the second signal sequence is an antibody signal sequence, optionally a human IgG2 or IgK signal sequence.

77. The rAAV genome of any one of claims 1-74, wherein the first signal sequence and/or the second signal sequence is a non-naturally occurring signal sequence.

78. The rAAV genome of any one of claims 1-74, wherein the first signal sequence and/or the second signal sequence comprises the amino acid sequence of SEQ ID No. 80.

79. The rAAV genome of any one of claims 1-74, wherein the first signal sequence and/or the second signal sequence comprises the amino acid sequence of SEQ ID No. 81.

80. The rAAV genome of any one of claims 1-79, wherein the first signal sequence comprises the amino acid sequence of SEQ ID No. 80 and the second signal sequence comprises the amino acid sequence of SEQ ID No. 81.

81. The rAAV genome of any one of claims 1-80, wherein the first coding sequence and/or the second coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 23, 96, 102, or 108.

82. The rAAV genome of any one of claims 1-80, wherein the first coding sequence and/or the second coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 24, 99, 105, 111, or 130.

83. The rAAV genome of any one of claims 1-82, wherein the first coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 23, 96, 102, or 108 and the second coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 24, 99, 105, 111, or 130.

84. The rAAV genome of any one of claims 1-83, wherein the antibody specifically binds to complement C5.

85. The rAAV genome of any one of claims 1-84, wherein the antibody heavy chain comprises the amino acid sequence of SEQ ID No. 64.

86. The rAAV genome of any one of claims 1-84, wherein the antibody heavy chain comprises the amino acid sequence of SEQ ID No. 82.

87. The rAAV genome of any one of claims 1-84, wherein the antibody light chain comprises the amino acid sequence of SEQ ID No. 77.

88. The rAAV genome of any one of claims 1-84, wherein the first coding sequence and/or the second coding sequence has been optimized for expression in a human cell.

89. The rAAV genome of any one of claims 1-84, wherein the first coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 52, 113, 114, or 115.

90. The rAAV genome of any one of claims 1-84, wherein the first coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 83, 94, 95, 101, or 107.

91. The rAAV genome of any one of claims 1-84, wherein the second coding sequence comprises any one of the nucleic acid sequences set forth in SEQ ID NOs 53, 98, 104, 110, or 131.

92. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 53.

93. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 63.

94. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 98.

95. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 99.

96. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 100.

97. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 104.

98. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 105.

99. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 106.

100. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 110.

101. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 111.

102. The rAAV genome of any one of claims 1-91, wherein the first coding sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs 52, 62, 83, 94, 95, 96, 97, 101, 102, 103, 107, 108, 109, 113, 114, and 115, and the second coding sequence comprises the nucleotide sequence set forth in SEQ ID NO 112.

103. The rAAV genome of any one of claims 1-102, wherein the rAAV genome is a single-stranded rAAV genome.

104. The rAAV genome of any one of claims 1-103, wherein the rAAV genome is a self-complementing rAAV genome.

105. The rAAV genome of any one of claims 1-104, wherein the rAAV genome comprises a nucleic acid sequence of SEQ ID No. 84.

106. The rAAV genome of any one of claims 1-104, wherein the rAAV genome comprises the nucleic acid sequence of SEQ ID NO: 85.

107. The rAAV genome of any one of claims 1-104, wherein the rAAV genome comprises the nucleic acid sequence of SEQ ID No. 86.

108. The rAAV genome of any one of claims 1-104, wherein the rAAV genome comprises the nucleic acid sequence of SEQ ID No. 87.

109. The rAAV genome of any one of claims 1-108, wherein the rAAV genome further comprises a 5 'inverted terminal repeat (5' itr) nucleotide sequence 5 'relative to the first polyadenylation sequence and a 3' inverted terminal repeat (3 'itr) nucleotide sequence 3' relative to the second polyadenylation sequence.

110. The rAAV genome of claim 109, wherein the 5'itr nucleotide sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence set forth in SEQ ID NO:14 and/or the 3' itr nucleotide sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence set forth in SEQ ID NO: 18.

111. The rAAV genome of any one of claims 1-110, wherein the rAAV genome comprises the nucleic acid sequence of SEQ ID No. 88.

112. The rAAV genome of any one of claims 1-110, wherein the rAAV genome comprises the nucleic acid sequence of SEQ ID No. 89.

113. The rAAV genome of any one of claims 1-110, wherein the rAAV genome comprises the nucleic acid sequence of SEQ ID No. 90.

114. The rAAV genome of any one of claims 1-110, wherein the rAAV genome comprises a nucleic acid sequence of SEQ ID No. 91.

115. A recombinant adeno-associated virus (rAAV), comprising:

(a) AAV capsids, the AAV capsids comprising AAV capsid proteins; and

(b) The rAAV genome of any one of the preceding claims.

116. The rAAV of claim 115, wherein the capsid protein is selected from the group consisting of: AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, and AAV9.

117. The rAAV of claim 115 or 116, wherein the AAV capsid protein comprises an amino acid sequence at least 95% identical to amino acid sequence of amino acids 203-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, or 17.

118. The rAAV of any one of claims 115-117, wherein: the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G.

119. The rAAV of claim 118, wherein:

120. The rAAV of claim 118, wherein the capsid protein comprises the amino acid sequence of amino acids 203-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, or 17.

121. The rAAV of any one of claims 115-120, wherein the AAV capsid protein comprises an amino acid sequence that is at least 95% identical to amino acid sequence of amino acids 138-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, or 17.

122. The rAAV of claim 121, wherein: the amino acid in the capsid protein corresponding to amino acid 151 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 160 of SEQ ID NO. 16 is D; the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G.

123. The rAAV of claim 122, wherein:

124. The rAAV of claim 122, wherein the capsid protein comprises the amino acid sequence of amino acids 138-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 15, 16, or 17.

125. The rAAV of any one of claims 115-124, wherein the AAV capsid protein comprises an amino acid sequence that is at least 95% identical to amino acid sequence of amino acids 1-736 of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, or 17.

126. The rAAV of claim 125, wherein: the amino acid in the capsid protein corresponding to amino acid 2 of SEQ ID NO. 16 is T; the amino acid in the capsid protein corresponding to amino acid 65 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 68 of SEQ ID NO. 16 is V; the amino acid in the capsid protein corresponding to amino acid 77 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 119 of SEQ ID NO. 16 is L; the amino acid in the capsid protein corresponding to amino acid 151 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 160 of SEQ ID NO. 16 is D; the amino acid in the capsid protein corresponding to amino acid 206 of SEQ ID NO. 16 is C; the amino acid in the capsid protein corresponding to amino acid 296 of SEQ ID NO. 16 is H; the amino acid in the capsid protein corresponding to amino acid 312 of SEQ ID NO. 16 is Q; the amino acid in the capsid protein corresponding to amino acid 346 of SEQ ID NO. 16 is A; the amino acid in the capsid protein corresponding to amino acid 464 of SEQ ID NO. 16 is N; the amino acid in the capsid protein corresponding to amino acid 468 of SEQ ID NO. 16 is S; the amino acid in the capsid protein corresponding to amino acid 501 of SEQ ID NO. 16 is I; the amino acid in the capsid protein corresponding to amino acid 505 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 590 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 626 of SEQ ID NO. 16 is G or Y; the amino acid in the capsid protein corresponding to amino acid 681 of SEQ ID NO. 16 is M; the amino acid in the capsid protein corresponding to amino acid 687 of SEQ ID NO. 16 is R; the amino acid in the capsid protein corresponding to amino acid 690 of SEQ ID NO. 16 is K; the amino acid in the capsid protein corresponding to amino acid 706 of SEQ ID NO. 16 is C; or the amino acid in the capsid protein corresponding to amino acid 718 of SEQ ID NO. 16 is G.

127. The rAAV of claim 126, wherein:

128. The rAAV of claim 127, wherein the capsid protein comprises the amino acid sequence of amino acids 1-736 of SEQ ID NOs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, or 17.

129. A polynucleotide comprising the nucleic acid sequence set forth in SEQ ID NO. 85-93.

130. A pharmaceutical composition comprising a rAAV according to any one of claims 115 to 128 or the polynucleotide of claim 129.

131. A packaging system for preparing a rAAV, wherein the packaging system comprises:

(a) A first nucleotide sequence encoding one or more AAV Rep proteins;

(b) A second nucleotide sequence encoding a capsid protein of a rAAV according to any one of claims 115 to 128; and

(c) A third nucleotide sequence comprising the rAAV genomic sequence of the rAAV of any one of claims 1 to 114.

132. The packaging system of claim 131, wherein the packaging system comprises a first vector comprising the first nucleotide sequence and the second nucleotide sequence, and a second vector comprising the third nucleotide sequence.

133. The packaging system of claim 131 or 132, further comprising a fourth nucleotide sequence comprising one or more helper virus genes.

134. The packaging system of claim 133, wherein the fourth nucleotide sequence is included within a third vector.

135. The packaging system of claim 133 or 134, wherein the fourth nucleotide sequence comprises one or more genes from a virus selected from the group consisting of: adenovirus, herpes virus, vaccinia virus and Cytomegalovirus (CMV).

136. The packaging system of any one of claims 131-135, wherein the first, second, and/or third vector is a plasmid.

137. A method for recombinantly producing a rAAV, the method comprising introducing the packaging system of any one of claims 131 to 135 into a cell under conditions whereby the rAAV is produced.

138. The rAAV of any one of claims 115-138, the pharmaceutical composition of claim 130, or the polynucleotide of claim 129 for use as a medicament.

139. The rAAV of any one of claims 115-138, the pharmaceutical composition of claim 130, or the polynucleotide of claim 129 for use in treating a complement C5-associated disease.

140. The rAAV of any one of claims 115-138, the pharmaceutical composition of claim 130, or the polynucleotide of claim 129 for use in a method of treating a subject having a complement C5-associated disease, the method comprising administering to the subject an effective amount of the rAAV, the pharmaceutical composition, or the polynucleotide.

141. A method of producing an antibody in a subject, the method comprising administering to the subject a pharmaceutical composition according to claim 130.

142. The method of claim 141, wherein the pharmaceutical composition is administered intravenously.

143. A method of treating a complement C5-associated disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the rAAV of any one of claims 115-128, the pharmaceutical composition of claim 130, or the polynucleotide of claim 129.

144. The method of claim 143, wherein the complement C5-associated disease is selected from the group consisting of: geographic Atrophy (GA), gillan-barre syndrome, myasthenia gravis, systemic Lupus Erythematosus (SLE) nephritis, proliferative nephritis, asthma, rheumatoid arthritis, sepsis, paroxysmal sleep hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), and age-related macular degeneration (AMD).

145. The method of claim 143 or 144, wherein the rAAV, the pharmaceutical composition, or the polynucleotide is administered intravenously.