WO2024062259A1 - Retroviral vector comprising rre inserted within an intron - Google Patents

Abstract

The present invention relates to retroviral vectors modified to improve transgene expression. In particular, the invention relates to retroviral vectors lacking an endogenous Rev response element (RRE) and comprising an intron, particularly a chimeric intron, into which an RRE has been inserted, as well as methods of production and uses thereof.

Description

RETROVIRAL VECTOR COMPRISING RRE INSERTED WITHIN AN INTRON

FIELD OF THE INVENTION

The present invention relates to retroviral vectors modified to improve transgene expression.

In particular, the invention relates to retroviral vectors lacking an endogenous Rev response element

(RRE) and comprising an intron, particularly a chimeric intron into which an RRE has been inserted, as well as methods of production and uses thereof.

BACKGROUND TO THE INVENTION

The use of nucleic acids as medicine, or gene therapy, is a promising new treatment modality.

The reason many gene therapies currently in use or under development are not effective at curing diseases is because it is difficult to make sufficient protein to reach the therapeutic threshold needed to treat or cure the disease. As such, generating sufficient gene expression is a major barrier to the success of many gene therapies.

The current approach to reaching the large doses needed for gene therapy to be successful is to administer massive amounts of the gene therapy to the patient, over 1 trillion viruses per kg of body mass. For example, Zolgensma is given at l.lxlO¹⁴ viral genomes per kg of body mass. Producing so much virus is expensive, contributing to the $ 1,000,000 USD cost of gene therapies, and giving so much virus to a person can trigger immune responses that threaten the health of the patient and the efficacy of the therapy. To circumvent these problems, research to-date has focused on gain of function mutations resulting in more potent proteins. Such an approach has been used previously in the gene therapies for haemophilia B (the Padua mutation in Factor IX) and lipoprotein lipase deficiency (the S447X variant of lipoprotein lipase). However, such gain-of-function mutations are not available for most gene therapies. Furthermore, even with gain-of-function mutations, high doses of the gene therapy vector were still necessary to make the treatment effective. Therefore, such gain- of-function mutations alone do not adequately address the existing problems associated with producing sufficient quantities of vector, or the unwanted and clinically dangerous side effects associated with the large doses required.

The present inventors have previously developed a lentiviral vector, which has been pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, comprising a promoter and a transgene. Typically, the backbone of the vector is from a simian immunodeficiency virus (SIV), such as SIV1 or African green monkey SIV (SIV-AGM). Preferably the backbone of a viral vector of the invention is from SIV-AGM. The HN and F proteins function, respectively, to attach to sialic acids and mediate cell fusion for vector entry to target cells. The present inventors discovered that this specifically F/HN-pseudotyped lentiviral vector can efficiently transduce airway epithelium, resulting in transgene expression sustained for periods beyond the proposed lifespan of airway epithelial cells. Importantly, the present inventors also found that re-administration does not result in a loss of efficacy. These features make the vectors of the present invention attractive candidates for treating diseases via their use in expressing therapeutic proteins: (i) within the cells of the respiratory tract; (ii) secreted into the lumen of the respiratory tract; and (iii) secreted into the circulatory system. However, even using this state-of-the-art platform technology, the levels of transgene expressed are at the lower predicted threshold required for clinical efficacy. There is therefore an unmet clinical need for new technologies to improve the potency of gene therapies. It is an object of the invention to address one or more of these problems. In particular, it is an object of the invention to provide new nucleic acid cassettes and gene therapy vectors which enable increased production of therapeutic proteins, allowing for lower doses of vector to be administered to patients. SUMMARY OF THE INVENTION At present, there remains a pressing need for technology that more efficiently produces therapeutic proteins for gene therapy, including from the inventors’ own lentiviral platform. To-date, other groups have focused on making each copy of a gene produce more of the therapeutic protein. For example, work on adeno-associated viral vectors (AAV vectors) have introduced an intron into the viral genome. The splicing of introns by a cell leads to the stabilization of mRNA, increasing the amount of mRNA in the cell and resulting in the production of more protein. However, this approach cannot readily be applied to viral vectors based on RNA viruses such as retrovirus and lentiviruses. This is because RNA genomes are subject to the same intron removal steps that mRNA is, such that introns are removed from RNA genomes during manufacturing, reducing the amount of protein an RNA viral vector can make. The present inventors have for the first time demonstrated that it is possible to introduce an intron into a lentiviral genome, and that the introduction of such an intron can increase transgene expression. In particular, the inventors have found that by removal of the endogenous Rev response element of a Simian Immunodeficiency Virus (SIV vector) pseudotyped with a VSV-G or F/HN envelope, and introduction of a β-globin/IgG intron with a precisely inserted SIV RRV into the SIV.VSV-G or SIV.F/HN genome increased AAT transgene expression by 686-fold or 501-fold, respectively, compared with corresponding vectors lacking an intron. The inventors’ innovative approach has the potential to provide several clinically important advantages: (i) allowing gene therapies to more easily reach the required therapeutic window, making them more efficacious; (ii) lowering the dose of a gene therapy agent required for administration to a patient, making the gene therapy safer; and/or (iii) lowering the production costs (as less vector is needed per patient), solving a major challenge for clinical trials, & pharmaceutical companies, and health care providers. Accordingly, the present invention provides a retroviral vector comprising an intron; wherein: (a) the endogenous Rev response element (RRE) of the retroviral genome is deleted; and (b) a retroviral RRE is inserted into the intron within 100 bp 5’ of the splice acceptor’s branch site. The retroviral RRE may be inserted within 20 bp 5’ of the splice acceptor’s branch site. The intron may be a chimeric intron, optionally selected from a β-globin/IgG chimeric intron or a chimeric intron from the CAGGS promoter. The intron may be a viral intron, optionally selected from SV40 intron, CMV Intron A and adenovirus tripartite leader sequence intron. The invention also preferably provides a retroviral vector comprising a chimeric intron; wherein: (a) the endogenous Rev response element (RRE) of the retroviral genome is deleted; and (b) a retroviral RRE is inserted into the chimeric intron. According to any retroviral vector of the invention, the retroviral RRE inserted into the intron may be the endogenous RRE of the retroviral genome. The RRE may be a Simian immunodeficiency virus (SIV) RRE. The RRE may comprise or consist of a nucleic acid sequence having at least 90% identity to SEQ ID NO: 1. According to any retroviral vector of the invention, the intron may be less than 1,000 bp in length, preferably less than 800 bp in length. According to any retroviral vector of the invention, the chimeric intron may be a β-globin/IgG chimeric intron or a chimeric intron from the CAGGS promoter. The chimeric intron may be a β- globin/IgG chimeric intron and the RRE inserted between (i) a splice donor site comprising or consisting of a nucleic acid sequence of TGAGTTTAAGGTAAGT (SEQ ID NO: 2); and (ii) a splice acceptor site comprising or consisting of a nucleic acid sequence of CTCTCCACAG (SEQ ID NO: 3). The β- globin/IgG chimeric intron may comprise or consist of a nucleic acid sequence having at least 90% identity to SEQ ID NO: 4. The intron may be a β-globin/IgG chimeric intron and the RRE an SIV RRE, and optionally wherein the chimeric intron comprising the RRE may comprise or consist of a nucleic acid sequence having at least 90% identity to SEQ ID NO: 5. According to any retroviral vector of the invention, the intron may be between a promoter and a transgene operably linked to said promoter, wherein optionally the promoter is selected from the group consisting of a cytomegalovirus (CMV) promoter, elongation factor 1a (EF1a) promoter, and a hybrid human CMV enhancer/EF1a (hCEF) promoter, preferably a hCEF promoter. The transgene may encode a therapeutic protein, wherein optionally said therapeutic protein is selected from: (a) a secreted therapeutic protein, optionally Alpha-1 Antitrypsin (AAT), Factor VIII, Surfactant Protein B (SFTPB), ADAMTS13, Factor VII, Factor IX, Factor X, Factor XI, von Willebrand Factor, Granulocyte- Macrophage Colony-Stimulating Factor (GM-CSF), Surfactant Protein C (SP-C), decorin, an anti- inflammatory protein and a monoclonal antibody against an infectious agent; or (b) CFTR, ABCA3, DNAH5, DNAH11, DNAI1, DNAI2, CSF2RA, CSF2RB and TRIM-72. Any retroviral vector of the invention may be a lentiviral vector. Said lentiviral vector may be selected from the group consisting of a Human immunodeficiency virus (HIV) vector, a Simian immunodeficiency virus (SIV) vector, a Feline immunodeficiency virus (FIV) vector, an Equine infectious anaemia virus (EIAV) vector, and a Visna/maedi virus vector. Any retroviral vector of the invention may be pseudotyped with haemagglutinin- neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus or G glycoprotein from Vesicular Stomatitis Virus (G-VSV). According to any retroviral vector of the invention, transgene expression may be increased by at least about 2-fold, preferably at least about 5-fold, more preferably at least about 10-fold compared with a corresponding vector which lacks a intron into which a retroviral RRE has been inserted. The invention also provides a nucleic acid comprising or consisting of a intron into which a retroviral RRE has been inserted, wherein optionally (i) the intron; and/or (ii) the RRE are as defined herein. The invention further provides a plasmid comprising a nucleic acid of the invention. The invention also provides a retroviral vector, nucleic acid and/or plasmid of the invention, which is codon-optimised. The invention further provides a composition comprising a retroviral vector, nucleic acid and/or plasmid of the invention, and a pharmaceutically-acceptable carrier. The invention also provides a host cell comprising a retroviral vector, nucleic acid and/or plasmid of the invention. The invention also provides a retroviral vector, nucleic acid plasmid or composition according as described herein for use in a method of treatment. The invention also provides a method of producing a retroviral vector, said method comprising the following steps: (a) growing cells in suspension; (b) transfecting the cells with one or more plasmids; (c) adding a nuclease; (d) harvesting the lentivirus; (e) adding trypsin; and (f) purification; wherein the one or more plasmids comprises a vector genome plasmid which comprises a nucleic acid of the invention, and optionally (i) a promoter of the invention and/or (ii) a transgene of the invention. The invention also provides a method of differentiating between a retroviral vector and a transgene expressed by said retroviral vector, said method comprising the steps of: (a1) transfecting cells with a retroviral vector of the invention; (b1) culturing the cells to allow transgene expression by the retrovirus; and (c1) quantifying RNA within the cells; or (a2) quantifying RNA within cells of a sample obtained from a patient who has undergone treatment with a retroviral vector, nucleic acid, plasmid or composition of the invention; wherein: (i) the amount of RNA comprising the chimeric intron into which a retroviral RRE has been inserted corresponds to the copy number of the retroviral vector; and (ii) the amount of RNA lacking the chimeric intron into which a retroviral RRE has been inserted corresponds to the amount of transgene mRNA; wherein optionally RNA is quantified by a PCR-based or in situ hybridisation-based assay. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: Rev response element (RRE) intron created by inserting rSIV RRE into a chimeric intron. (A) Schematic of the chimeric intron which is composed of the splice donor from an intron in Hemoglobin subunit B (black), and the splice acceptor from an intron in immunoglobulin gamma (grey). (B) Schematic of the RRE intron, where the rSIV RRE (light grey) was inserted between the splice donor and splice acceptor. Figure 2: A-H show schematic drawings of exemplary plasmids used for production of the vectors of the invention. (A) Shows a schematic of an intron containing lentiviral vector genome plasmid (pDNA1) encoding an Alpha-1-Antitrypsin transgene. An intron has been inserted between the promoter (hCEF) and the RRE has been moved to within the intron. (B) Shows a schematic of a plasmid encoding codon optimized SIV Gag and Pol (pDNA2a) for lentiviral production. (C) Shows a schematic of a plasmid encoding SIV Gag and Pol (pDNA2a) for lentiviral production. (D) Shows a schematic of a plasmid encoding SIV Rev for lentiviral production (pDNA2b). (E) Shows a schematic of a plasmid encoding the fusion protein from Sendai virus (pDNA3a) for lentiviral production. (F) Shows a schematic of a plasmid encoding the hemagglutinin-neuraminidase protein from Sendai virus (pDNA3b) for lentiviral production. (G) Shows a schematic of a no intron lentiviral vector genome plasmid encoding an Alpha- 1-Antitrypsin transgene (pDNA1). The RRE element is 5ʹ of the promoter (hCEF) between the Partial GAG and cPPT sequences. (H) Shows a schematic of a plasmid encoding the VSV glycoprotein (pDNA3) for lentiviral production. Figure 3: RRE Intron enhances AAT expression 10.6x. HEK293T cells were transfected with a plasmid encoding an Alpha-1-Antitrypsin (AAT) transgene without (No Intron) or with (Intron) the RRE intron. Inclusion of the intron significantly enhanced AAT expression (p = 0.0003). Neg – negative control. Each dot represents a different well of transduced HEK293T cells. A Mann-Whitney test was used for statistical analysis. Figure 4: RRE Intron is correctly spliced in HEK293T cells. DNA (A) and RNA (B) was extracted from HEK293Tcells transfected with plasmids encoding an Alpha-1-Antitrypsin (AAT) transgene without (pGM407 - No Intron) or with (pGM991 - Intron) the RRE intron. (A) PCR of DNA extracted from transfected cells verified that pGM991 contains the 760 bp RRE intron. (B) Reverse Transcriptase PCR of RNA extracted from transfected cells verified that the RRE intron was spliced during mRNA maturation. Figure 5: RRE intron is packaged into rSIV.VSV-G lentivirus. DNA was extracted from HEK293T cells transduced with rSIV.VSV-G lentivirus expressing AAT without (vGM290) and with (vGM291) an RRE intron. Non-transduced cells (NTC) were used as controls. PCR of the resulting DNA using primers that bind to either side of the intron reveal that the intron was packaged into vGM291, producing an 1127 bp product, whereas in the absence of an intron, vGM290 produces a smaller 371 bp product. A no template control (n) as well as the lentiviral transfer plasmids for vGM290 (pGM407) and vGM291 (pGM991) were included. Figure 6: RRE intron is successfully spliced by transduced HEK293T cells. RNA was extracted from HEK293T cells transduced with rSIV.VSV-G lentivirus expressing AAT without (vGM290) and with (vGM291) an RRE intron. Non-transduced cells (NTC) were used as controls. RT-PCR of the resulting RNA using primers that bind to either side of the intron reveal that the intron was spliced during expression of vGM291, producing the same 277 bp produced cells transduced with vGM290. A no template control (n) as well as the lentiviral transfer plasmids for vGM290 (pGM407) and vGM291 (pGM991) were included. Figure 7: RRE Intron enhances AAT expression 686-fold in HEK293T cells. HEK293T cells were transduced with rSIV.VSV-G lentivirus expressing AAT without (No Intron) and with (Intron) an RRE intron. Compared to the state of the art no intron virus, the RRE containing virus significantly enhanced AAT expression (p=0.0037, Kruskal-Wallis test with Dunn’s multiple comparison correction). Non- transduced cells (NTC) were used as controls. Figure 8: An RRE Intron increases transgene transcription in HEK293T cells. (A) RT-ddPCR was performed on RNA extracted from transfected HEK293T cells. Inclusion of the intron increased the amount of mRNA (WPRE copies). WPRE transcript copies were standardized to the house keeping gene Beta-2-Microglobulin (B2M) (B). Inclusion of an RRE intron increases the amount of mRNA produced per plasmid copy (quantified by ddPCR from DNA extracted from transfected HEK293T cells). Each dot represents a different well of transduced HEK293T cells. Statistical analysis was performed with a Mann-Whitney test. Figure 9: RRE Intron enhances AAT expression 501-fold in HEK293T cells. HEK293T cells were transduced with rSIV.F/HN lentivirus expressing AAT without (No Intron) and with (Intron) an RRE intron. Compared to the state of the art no intron virus, the RRE containing virus significantly enhanced AAT expression (p=0.0002, Mann-Whitney test). Non-transduced cells (NTC) were used as controls. DETAILED DESCRIPTION OF THE INVENTION Definitions Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Singleton, et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 20 ED., John Wiley and Sons, New York (1994), and Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY, Harper Perennial, NY (1991) provide the skilled person with a general dictionary of many of the terms used in this disclosure. The meaning and scope of the terms should be clear; however, in the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. This disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of this disclosure. The terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims. Unless otherwise indicated, any nucleic acid sequences are written left to right in 5' to 3' orientation; amino acid sequences are written left to right in amino to carboxy orientation, respectively. The headings provided herein are not limitations of the various aspects or embodiments of this disclosure. As used herein, the term "capable of' when used with a verb, encompasses or means the action of the corresponding verb. For example, "capable of interacting" also means interacting, "capable of cleaving" also means cleaves, "capable of binding" also means binds and "capable of specifically targeting…" also means specifically targets. Other definitions of terms may appear throughout the specification. Before the exemplary embodiments are described in more detail, it is to be understood that this disclosure is not limited to particular embodiments described, and as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be defined only by the appended claims. Numeric ranges are inclusive of the numbers defining the range. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within this disclosure. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within this disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in this disclosure. As used herein, the articles "a" and “an” may refer to one or to more than one (e.g. to at least one) of the grammatical object of the article. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. In this application, the use of "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "including", as well as other forms, such as "includes" and "included", is not limiting. “About” may generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5% of a given value or range of values. Preferably, the term “about” shall be understood herein as plus or minus (±) 5%, preferably ± 4%, ± 3%, ± 2%, ± 1%, ± 0.5%, ± 0.1%, of the numerical value of the number with which it is being used. The term "consisting of'' refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the invention. As used herein the term "consisting essentially of'' refers to those elements required for a given invention. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that invention (i.e. inactive or non-immunogenic ingredients). Embodiments described herein as “comprising” one or more features may also be considered as disclosure of the corresponding embodiments “consisting of” and/or “consisting essentially of” such features. Concentrations, amounts, volumes, percentages and other numerical values may be presented herein in a range format. It is also to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. A "vector" or "construct" (sometimes referred to as gene delivery or gene transfer "vehicle") refers to a macromolecule or complex of molecules comprising a polynucleotide to be delivered to a host cell, either in vitro or in vivo. A vector can be a linear or a circular molecule. A vector of the invention may be viral or non-viral. All disclosure herein in relation vectors of the invention applies equally to viral and non-viral vectors unless otherwise stated. All disclosure in relation to viral vectors of the invention applies equally and without reservation to lentiviral (e.g. SIV) vectors, particularly to lentiviral (e.g. SIV) vectors that are pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus (also referred to herein as SIV F/HN or SIV-FHN). As used herein, the terms "viral vector”, “retroviral vector” and “retroviral F/HN vector” are used interchangeably to mean a retroviral vector comprising a retroviral RNA sequence and pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, unless otherwise stated. The terms “lentiviral vector” and “lentiviral F/HN vector” are used interchangeably to mean a lentiviral vector pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, unless otherwise stated. All disclosure herein in relation to retroviral vectors of the invention applies equally and without reservation to lentiviral vectors of the invention and to SIV vectors that are pseudotyped with hemagglutinin- neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus (also referred to herein as SIV F/HN or SIV-FHN). The term “intron” as used herein refers to a nucleic acid sequence within a gene that is located between exons. Introns are transcribed along with the exons but are removed from the primary gene transcript by RNA splicing to leave mature mRNA. The removal of introns typically leads to the stabilization of mRNA, increasing the amount of mRNA in the cell. Rev (regulator of virion) is a trans-acting nuclear protein whose functional expression are required for retroviral replication. Specifically, the rev gene products are required for processing and translation of the gag and env mRNAs, and thus rev regulates the expression of the viral structural proteins. The term “Rev-responsive element” (RRE) refers to a cis-acting anti-repression sequence in env, which is responsive to the rev gene product. mRNAs that contain an RRE can be exported from the nucleus to the cytoplasm for translation and virion packaging. The terms “RRE” and “RRE sequence” are used interchangeably herein. As used herein, the term "plasmid", refers to a common type of non-viral vector. A plasmid is an extra-chromosomal DNA molecule separate from the chromosomal DNA which is capable of replicating independently of the chromosomal DNA. Preferably a plasmid is circular and may be double-stranded. The terms "nucleic acid cassette”, “nucleic acid construct", "expression cassette" and "nucleic acid expression cassette" are used interchangeably to mean a nucleic acid molecule that is capable of directing transcription. A nucleic acid cassette includes, at the least, a promoter or a structure functionally equivalent to a promoter and a nucleic acid sequence to be transcribed. Thus, a nucleic acid cassette includes, at the least, a promoter or a structure functionally equivalent to a promoter and a nucleic acid sequence encoding a protein of interest. In the present invention, a nucleic acid cassette includes, at the least, a promoter or a structure functionally equivalent to a promoter, a nucleic acid sequence encoding a signal peptide and a nucleic acid encoding a therapeutic protein. A nucleic acid cassette may include additional elements, such as an enhancer, and/or a transcription termination signal. As used herein the terms “signal peptide”, “signal sequence”, “targeting sequence”, “leader sequence” and “secretory signal” are used interchangeably to mean heterogenous peptide sequences that are found at the N-terminus of secreted proteins that are instrumental in initiating the secretion process. In particular, signal peptides are found in proteins that are targeted to the endoplasmic reticulum and eventually destined to be either secreted or retained in the cell membrane of the cell, particularly as single-pass membrane proteins. Signal peptides are typically removed to produce the mature form of the protein. Signal peptides are normally short peptides, typically about 5 to about 40 amino acids in length, such as about 5 to about 35, or about 10 to about 35 amino acids in length, preferably about 10 to about 30 or about 15 to about 30 amino acids in length. A signal peptide may comprise a core of hydrophobic amino acids, said core typically being about 4 to about 20, such as about 5 to about 20, about 5 to about 16 or about 5 to about 15 amino acids in length). When present, a signal peptide is typically present at the N-terminus of a protein. As used herein, the terms “transduced” and “modified” are used interchangeably to describe cells which have been modified to express a transgene of interest. Typically the modification occurs through transduction of the cells. The term "endogenous," when used in relation to a Rev response element (RRE) refers to the RRE which is from the same retroviral/lentiviral vector as the retroviral/lentiviral vector of the invention. A wildtype/unmodified vector will comprise an RRE within its genome, typically at a defined standard location. Viral vectors of the present invention typically have genomes which lack their endogenous RRE. The endogenous RRE may be inserted into an intron which is then itself introduced into the retroviral/lentiviral genome. An exogenous RRE is from a different virus to the viral vector of the invention. By way of non- limiting example, the viral vector may be an HIV vector, and the RRE may be a SIV RRE. As used herein, the terms “titre” and “yield” are used interchangeably to mean the amount of lentiviral (e.g. SIV) vector produced by a method of the invention. Titre is the primary benchmark characterising manufacturing efficiency, with higher titres generally indicating that more retroviral/lentiviral (e.g. SIV) vector is manufactured (e.g. using the same amount of reagents). Titre or yield may relate to the number of vector genomes that have integrated into the genome of a target cell (integration titre), which is a measure of “active” virus particles, i.e. the number of particles capable of transducing a cell. Transducing units (TU/mL also referred to as TTU/mL) is a biological readout of the number of host cells that get transduced under certain tissue culture/virus dilutions conditions, and is a measure of the number of “active” virus particles. The total number of (active+inactive) virus particles may also be determined using any appropriate means, such as by measuring either how much Gag is present in the test solution or how many copies of viral RNA are in the test solution. Assumptions are then made that a lentivirus particle contains either 2000 Gag molecules or 2 viral RNA molecules. Once total particle number and a transducing titre/TU have been measured, a particle:infectivity ratio calculated. Amino acids are referred to herein using the name of the amino acid, the three-letter abbreviation or the single letter abbreviation. As used herein, the terms "protein" and "polypeptide" are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha-amino and carboxyl groups of adjacent residues. The terms "protein", and "polypeptide" refer to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogues, regardless of its size or function. "Protein" and "polypeptide" are often used in reference to relatively large polypeptides, whereas the term "peptide" is often used in reference to small polypeptides, but usage of these terms in the art overlaps. The terms "protein" and "polypeptide" are used interchangeably herein when referring to a gene product and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogues of the foregoing. As used herein, the terms “polynucleotides”, "nucleic acid" and "nucleic acid sequence" refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analogue thereof. The nucleic acid can be either single-stranded or double-stranded. A single-stranded nucleic acid can be one nucleic acid strand of a denatured double- stranded DNA Alternatively, it can be a single-stranded nucleic acid not derived from any double- stranded DNA. In one aspect, the nucleic acid can be DNA. In another aspect, the nucleic acid can be RNA Suitable nucleic acid molecules are DNA, including genomic DNA or cDNA. Other suitable nucleic acid molecules are RNA, including siRNA, shRNA, and antisense oligonucleotides. The terms “transgene” and “gene” are also used interchangeably and both terms encompass fragments or variants thereof encoding the target protein. The transgenes of the present invention include nucleic acid sequences that have been removed from their naturally occurring environment, recombinant or cloned DNA isolates, and chemically synthesized analogues or analogues biologically synthesized by heterologous systems. Minor variations in the amino acid sequences of the invention are contemplated as being encompassed by the present invention, providing that the variations in the amino acid sequence(s) maintain at least 60%, at least 70%, more preferably at least 80%, at least 85%, at least 90%, at least 95%, and most preferably at least 97% or at least 99% sequence identity to the amino acid sequence of the invention or a fragment thereof as defined anywhere herein. The term homology is used herein to mean identity. As such, the sequence of a variant or analogue sequence of an amino acid sequence of the invention may differ on the basis of substitution (typically conservative substitution) deletion or insertion. Proteins comprising such variations are referred to herein as variants. Proteins of the invention may include variants in which amino acid residues from one species are substituted for the corresponding residue in another species, either at the conserved or non- conserved positions. Variants of protein molecules disclosed herein may be produced and used in the present invention. Following the lead of computational chemistry in applying multivariate data analysis techniques to the structure/property-activity relationships [see for example, Wold, et al. Multivariate data analysis in chemistry. Chemometrics-Mathematics and Statistics in Chemistry (Ed.: B. Kowalski); D. Reidel Publishing Company, Dordrecht, Holland, 1984 (ISBN 90-277-1846-6] quantitative activity-property relationships of proteins can be derived using well-known mathematical techniques, such as statistical regression, pattern recognition and classification [see for example Norman et al. Applied Regression Analysis. Wiley-lnterscience; 3rd edition (April 1998) ISBN: 0471170828; Kandel, Abraham et al. Computer-Assisted Reasoning in Cluster Analysis. Prentice Hall PTR, (May 11, 1995), ISBN: 0133418847; Krzanowski, Wojtek. Principles of Multivariate Analysis: A User's Perspective (Oxford Statistical Science Series, No 22 (Paper)). Oxford University Press; (December 2000), ISBN: 0198507089; Witten, Ian H. et al Data Mining: Practical Machine Learning Tools and Techniques with Java Implementations. Morgan Kaufmann; (October 11, 1999), ISBN:1558605525; Denison David G. T. (Editor) et al Bayesian Methods for Nonlinear Classification and Regression (Wiley Series in Probability and Statistics). John Wiley & Sons; (July 2002), ISBN: 0471490369; Ghose, Arup K. et al. Combinatorial Library Design and Evaluation Principles, Software, Tools, and Applications in Drug Discovery. ISBN: 0-8247-0487-8]. The properties of proteins can be derived from empirical and theoretical models (for example, analysis of likely contact residues or calculated physicochemical property) of proteins sequence, functional and three-dimensional structures and these properties can be considered individually and in combination. Amino acids are referred to herein using the name of the amino acid, the three-letter abbreviation or the single letter abbreviation. The term “protein", as used herein, includes proteins, polypeptides, and peptides. As used herein, the term “amino acid sequence” is synonymous with the term “polypeptide” and/or the term “protein”. In some instances, the term “amino acid sequence” is synonymous with the term “peptide”. The terms "protein" and "polypeptide" are used interchangeably herein. In the present disclosure and claims, the conventional one-letter and three- letter codes for amino acid residues may be used. The 3-letter code for amino acids as defined in conformity with the IUPACIUB Joint Commission on Biochemical Nomenclature (JCBN). It is also understood that a polypeptide may be coded for by more than one nucleotide sequence due to the degeneracy of the genetic code. Amino acid residues at non-conserved positions may be substituted with conservative or non- conservative residues. In particular, conservative amino acid replacements are contemplated. A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, or histidine), acidic side chains (e.g., aspartic acid or glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, or cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, or tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, or histidine). Thus, if an amino acid in a polypeptide is replaced with another amino acid from the same side chain family, the amino acid substitution is considered to be conservative. The inclusion of conservatively modified variants in a protein of the invention does not exclude other forms of variant, for example polymorphic variants, interspecies homologs, and alleles. “Non-conservative amino acid substitutions” include those in which (i) a residue having an electropositive side chain (e.g., Arg, His or Lys) is substituted for, or by, an electronegative residue (e.g., Glu or Asp), (ii) a hydrophilic residue (e.g., Ser or Thr) is substituted for, or by, a hydrophobic residue (e.g., Ala, Leu, Ile, Phe or Val), (iii) a cysteine or proline is substituted for, or by, any other residue, or (iv) a residue having a bulky hydrophobic or aromatic side chain (e.g., Val, His, Ile or Trp) is substituted for, or by, one having a smaller side chain (e.g., Ala or Ser) or no side chain (e.g., Gly). “Insertions” or “deletions” are typically in the range of about 1, 2, or 3 amino acids. The variation allowed may be experimentally determined by systematically introducing insertions or deletions of amino acids in a protein using recombinant DNA techniques and assaying the resulting recombinant variants for activity. This does not require more than routine experiments for a skilled person. A “fragment” of a polypeptide comprises at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or more of the original polypeptide. The polynucleotides of the present invention may be prepared by any means known in the art. For example, large amounts of the polynucleotides may be produced by replication in a suitable host cell. The natural or synthetic DNA fragments coding for a desired fragment will be incorporated into recombinant nucleic acid constructs, typically DNA constructs, capable of introduction into and replication in a prokaryotic or eukaryotic cell. Usually the DNA constructs will be suitable for autonomous replication in a unicellular host, such as yeast or bacteria, but may also be intended for introduction to and integration within the genome of a cultured insect, mammalian, plant or other eukaryotic cell lines. The polynucleotides of the present invention may also be produced by chemical synthesis, e.g. by the phosphoramidite method or the tri-ester method, and may be performed on commercial automated oligonucleotide synthesizers. A double-stranded fragment may be obtained from the single stranded product of chemical synthesis either by synthesizing the complementary strand and annealing the strand together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence. When applied to a nucleic acid sequence, the term “isolated” in the context of the present invention denotes that the polynucleotide sequence has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences (but may include naturally occurring 5' and 3' untranslated regions such as promoters and terminators), and is in a form suitable for use within genetically engineered protein production systems. Such isolated molecules are those that are separated from their natural environment. In view of the degeneracy of the genetic code, considerable sequence variation is possible among the polynucleotides of the present invention. Degenerate codons encompassing all possible codons for a given amino acid are set forth below: Amino Acid Codons Degenerate Codon Cys TGC TGT TGY Ser AGC AGT TCA TCC TCG TCT WSN Thr ACA ACC ACG ACT ACN Pro CCA CCC CCG CCT CCN Ala GCA GCC GCG GCT GCN Gly GGA GGC GGG GGT GGN Asn AAC AAT AAY Asp GAC GAT GAY Glu GAA GAG GAR Gln CAA CAG CAR His CAC CAT CAY Arg AGA AGG CGA CGC CGG CGT MGN Lys AAA AAG AAR Met ATG ATG Ile ATA ATC ATT ATH Leu CTA CTC CTG CTT TTA TTG YTN Val GTA GTC GTG GTT GTN Phe TTC TTT TTY Tyr TAC TAT TAY Trp TGG TGG Ter TAA TAG TGA TRR Asn/ Asp RAY Glu/ Gln SAR Any NNN One of ordinary skill in the art will appreciate that flexibility exists when determining a degenerate codon, representative of all possible codons encoding each amino acid. For example, some polynucleotides encompassed by the degenerate sequence may encode variant amino acid sequences, but one of ordinary skill in the art can easily identify such variant sequences by reference to the amino acid sequences of the present invention. A “variant” nucleic acid sequence has substantial homology or substantial similarity to a reference nucleic acid sequence (or a fragment thereof). A nucleic acid sequence or fragment thereof is “substantially homologous” (or “substantially identical”) to a reference sequence if, when optimally aligned (with appropriate nucleotide insertions or deletions) with the other nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 70%, 75%, 80%, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or more% of the nucleotide bases. Methods for homology determination of nucleic acid sequences are known in the art. Alternatively, a “variant” nucleic acid sequence is substantially homologous with (or substantially identical to) a reference sequence (or a fragment thereof) if the “variant” and the reference sequence they are capable of hybridizing under stringent (e.g. highly stringent) hybridization conditions. Nucleic acid sequence hybridization will be affected by such conditions as salt concentration (e.g. NaCl), temperature, or organic solvents, in addition to the base composition, length of the complementary strands, and the number of nucleotide base mismatches between the hybridizing nucleic acids, as will be readily appreciated by those skilled in the art. Stringent temperature conditions are preferably employed, and generally include temperatures in excess of 30°C, typically in excess of 37°C and preferably in excess of 45°C. Stringent salt conditions will ordinarily be less than 1000 mM, typically less than 500 mM, and preferably less than 200 mM. The pH is typically between 7.0 and 8.3. The combination of parameters is much more important than any single parameter. Methods of determining nucleic acid percentage sequence identity are known in the art. By way of example, when assessing nucleic acid sequence identity, a sequence having a defined number of contiguous nucleotides may be aligned with a nucleic acid sequence (having the same number of contiguous nucleotides) from the corresponding portion of a nucleic acid sequence of the present invention. Tools known in the art for determining nucleic acid percentage sequence identity include Nucleotide BLAST (as described below). One of ordinary skill in the art appreciates that different species exhibit “preferential codon usage”. As used herein, the term “preferential codon usage” refers to codons that are most frequently used in cells of a certain species, thus favouring one or a few representatives of the possible codons encoding each amino acid. For example, the amino acid threonine (Thr) may be encoded by ACA, ACC, ACG, or ACT, but in mammalian host cells ACC is the most commonly used codon; in other species, different codons may be preferential. Preferential codons for a particular host cell species can be introduced into the polynucleotides of the present invention by a variety of methods known in the art. Introduction of preferential codon sequences into recombinant DNA can, for example, enhance production of the protein by making protein translation more efficient within a particular cell type or species. Thus, according to the invention, in addition to the gag-pol genes any nucleic acid sequence may be codon-optimised for expression in a host or target cell. In particular, the vector genome (or corresponding plasmid), the REV gene (or corresponding plasmid), the fusion protein (F) gene (or correspond plasmid) and/or the hemagglutinin-neuraminidase (HN) gene (or corresponding plasmid, or any combination thereof may be codon-optimised. A “fragment” of a polynucleotide of interest comprises a series of consecutive nucleotides from the sequence of said full-length polynucleotide. By way of example, a “fragment” of a polynucleotide of interest may comprise (or consist of) at least 30 consecutive nucleotides from the sequence of said polynucleotide (e.g. at least 35, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800 850, 900, 950 or 1000 consecutive nucleic acid residues of said polynucleotide). A fragment may include at least one antigenic determinant and/or may encode at least one antigenic epitope of the corresponding polypeptide of interest. Typically, a fragment as defined herein retains the same function as the full-length polynucleotide. The terms "decrease", "reduced", "reduction", or "inhibit" are all used herein to mean a decrease by a statistically significant amount. The terms "reduce," "reduction" or "decrease" or "inhibit" typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99% , or more. As used herein, "reduction" or "inhibition" encompasses a complete inhibition or reduction as compared to a reference level. "Complete inhibition" is a 100% inhibition (i.e. abrogation) as compared to a reference level. The terms "increased", "increase", "enhance", or "activate" are all used herein to mean an increase by a statically significant amount. The terms "increased", "increase", "enhance", or "activate" can mean an increase of at least 25%, at least 50% as compared to a reference level, for example an increase of at least about 50%, or at least about 75%, or at least about 80%, or at least about 90%, or at least about 100%, or at least about 150%, or at least about 200%, or at least about 250% or more compared with a reference level, or at least about a 1.5-fold, or at least about a 2-fold, or at least about a 2.5-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 1.5-fold and 10-fold or greater as compared to a reference level. In the context of a yield or titre, an "increase" is an observable or statistically significant increase in such level. The terms "individual”, "subject”, and "patient”, are used interchangeably herein to refer to a mammalian subject for whom diagnosis, prognosis, disease monitoring, treatment, therapy, and/or therapy optimisation is desired. The mammal can be (without limitation) a human, non-human primate, mouse, rat, dog, cat, horse, or cow. In a preferred embodiment, the individual, subject, or patient is a human. An “individual” may be an adult, juvenile or infant. An “individual” may be male or female. A "subject in need" of treatment for a particular condition can be an individual having that condition, diagnosed as having that condition, or at risk of developing that condition. A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment or one or more complications or symptoms related to such a condition, and optionally, have already undergone treatment for a condition as defined herein or the one or more complications or symptoms related to said condition. Alternatively, a subject can also be one who has not been previously diagnosed as having a condition as defined herein or one or more or symptoms or complications related to said condition. For example, a subject can be one who exhibits one or more risk factors for a condition, or one or more or symptoms or complications related to said condition or a subject who does not exhibit risk factors. As used herein, the term “healthy individual” refers to an individual or group of individuals who are in a healthy state, e.g. individuals who have not shown any symptoms of the disease, have not been diagnosed with the disease and/or are not likely to develop the disease e.g. cystic fibrosis (CF) or any other disease described herein). Preferably said healthy individual(s) is not on medication affecting CF and has not been diagnosed with any other disease. The one or more healthy individuals may have a similar sex, age, and/or body mass index (BMI) as compared with the test individual. Application of standard statistical methods used in medicine permits determination of normal levels of expression in healthy individuals, and significant deviations from such normal levels. Herein the terms “control” and “reference population” are used interchangeably. The term “pharmaceutically acceptable” as used herein means approved by a regulatory agency of the Federal or a state government, or listed in the U.S. Pharmacopeia, European Pharmacopeia or other generally recognized pharmacopeia The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that such publications constitute prior art to the claims appended hereto. Disclosure related to the various methods of the invention are intended to be applied equally to other methods, therapeutic uses or methods, the data storage medium or device, the computer program product, and vice versa. Retroviral and Lentiviral Vectors The invention relates to a retroviral/lentiviral (e.g. SIV) vector. Retroviral/lentiviral vectors of the invention, can integrate into the genome of transduced cells and lead to long-lasting expression. The term “retrovirus” refers to any member of the Retroviridae family of RNA viruses that encode the enzyme reverse transcriptase. The term “lentivirus” refers to a family of retroviruses. Thus, all references herein to retroviral vectors of the invention apply equally and without reservation to lentiviral vectors. Further, all references herein to lentiviral vectors of the invention apply equally and without reservation to retroviral vectors. Examples of retroviruses suitable for use in the present invention include gamma retroviruses such as murine leukaemia virus (MLV) and feline leukaemia virus (FLV). Examples of lentiviruses suitable for use in the present invention include Simian immunodeficiency virus (SIV), Human immunodeficiency virus (HIV), Feline immunodeficiency virus (FIV), Equine infectious anaemia virus (EIAV), and Visna/maedi virus. Preferably the invention relates to lentiviral vectors and the production thereof. A particularly preferred lentiviral vector is an SIV vector (including all strains and subtypes), such as a SIV-AGM (originally isolated from African green monkeys, Cercopithecus aethiops). Alternatively the invention relates to HIV vectors. The retroviral/lentiviral (e.g. SIV) vectors of the present invention are typically pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, or with G glycoprotein from Vesicular Stomatitis Virus (referred to as VSV-G or G-VSV). Preferably the lentiviral (e.g. SIV) vectors of the present invention are pseudotyped with HN and F from a respiratory paramyxovirus. Particularly preferably the respiratory paramyxovirus is a Sendai virus (murine parainfluenza virus type 1). The retroviral/lentiviral (e.g. SIV) vectors of the present invention may be pseudotyped with proteins from another virus, provided that the pseudotyping proteins do not negatively impact the manufactured titre of the vector (or even result in an increased titre of the vector) and/or transgene expression (or even result in increased transgene expression). Non-limiting examples of other proteins that may be used to pseudotype retroviral/lentiviral (e.g. SIV) vectors of the present invention include severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein or modified forms thereof. VSV-G and SARS-Cov2 spike protein used for pseudotyping are as those described in UK Patent Application Nos. 2118685.3 and International Application No. PCT/GB2022/050933, each of which is herein incorporated by reference in its entirety. A retroviral/lentiviral (e.g. SIV) vector for use according to the invention may be integrase- competent (IC). Alternatively, the lentiviral (e.g. SIV) vector may be integrase-deficient (ID). Viral vectors of the invention, particularly retroviral/lentiviral (e.g. SIV) vectors as described herein may transduce one or more cell types as described herein to achieve long term transgene expression. The retroviral/lentiviral (e.g. SIV) vectors of the present invention enable high levels of transgene expression. In particular, retroviral/lentiviral (e.g. SIV) vectors of the present invention typically result in high levels (therapeutic levels) of expression of a therapeutic protein. The nucleic acid sequence encoding a therapeutic protein to be included in a viral vector of the invention, particularly a retroviral/lentiviral (e.g. SIV) vector of the invention may be modified to facilitate expression. For example, the transgene sequence may be in CpG-depleted (or CpG-fee) and/or codon-optimised form to facilitate gene expression. Standard techniques for modifying the transgene sequence in this way are known in the art. The genome of the retroviral/lentiviral (e.g. SIV) vector may be fully or partially CpG-depleted (or CpG-fee) and/or codon-optimised. Retroviral/lentiviral (e.g. SIV) vectors, such as those of the invention, can integrate into the genome of transduced cells and lead to long-lasting expression, making them suitable for transduction of stem/progenitor cells. In the lung, several cell types with regenerative capacity have been identified as responsible for maintaining specific cell lineages in the conducting airways and alveoli. These include basal cells and submucosal gland duct cells in the upper airways, club cells and neuroendocrine cells in the bronchiolar airways, bronchioalveolar stem cells in the terminal bronchioles and type II pneumocytes in the alveoli. Therefore, and without being bound by theory, it is believed that said retroviral/lentiviral (e.g. SIV) vectors bring about long term gene expression of the transgene of interest by introducing the transgene into one or more long-lived airway epithelial cells or cell types, such as basal cells and submucosal gland duct cells in the upper airways, club cells and neuroendocrine cells in the bronchiolar airways, bronchioalveolar stem cells in the terminal bronchioles type II pneumocytes in the alveoli, submucosal acinar cells, ionocytes, and type I pneumocytes. As demonstrated herein, the integration of retroviral/lentiviral (e.g. SIV) vectors with modified retroviral/lentiviral (e.g. SIV) RNA sequences of the invention into target cell genomes is unexpectedly not negatively impacted, and in fact may even be increased. Accordingly, the retroviral/lentiviral (e.g. SIV) vectors of the invention may transduce one or more cells or cell lines with regenerative potential within the lung (including the airways and respiratory tract) to achieve long term gene expression. For example, the retroviral/lentiviral (e.g. SIV) vectors may transduce basal cells, such as those in the upper airways/respiratory tract. Basal cells have a central role in processes of epithelial maintenance and repair following injury. In addition, basal cells are widely distributed along the human respiratory epithelium, with a relative distribution ranging from 30% (larger airways) to 6% (smaller airways). The retroviral/lentiviral (e.g. SIV) vectors of the invention may be used to transduce isolated and expanded stem/progenitor cells ex vivo prior administration to a patient. Preferably, the retroviral/lentiviral (e.g. SIV) vectors of the invention are used to transduce cells within the lung (or airways/respiratory tract) in vivo. The retroviral/lentiviral (e.g. SIV) vectors of the invention demonstrate remarkable resistance to shear forces with only modest reduction in transduction ability when passaged through clinically- relevant delivery devices such as bronchoscopes, spray bottles and nebulisers. The retroviral/lentiviral (e.g. SIV) vectors of the present invention enable high levels of transgene expression, resulting in high levels (therapeutic levels) of expression of a therapeutic protein. The retroviral/lentiviral (e.g. SIV) vectors of the present invention typically provide high expression levels of a transgene when administered to a patient. The terms high expression and therapeutic expression are used interchangeably herein. Expression may be measured by any appropriate method (qualitative or quantitative, preferably quantitative), and concentrations given in any appropriate unit of measurement, for example ng/ml or nM. Expression of a transgene of interest may be given relative to the expression of the corresponding endogenous (defective) gene in a patient. Expression may be measured in terms of mRNA or protein expression. The expression of the transgene of the invention, such as a functional CFTR gene, may be quantified relative to the endogenous gene, such as the endogenous (dysfunctional) CFTR genes in terms of mRNA copies per cell or any other appropriate unit. Expression levels of a transgene and/or the encoded therapeutic protein of the invention may be measured in the lung tissue, epithelial lining fluid and/or serum/plasma as appropriate. A high and/or therapeutic expression level may therefore refer to the concentration in the lung, epithelial lining fluid and/or serum/plasma. A retroviral/lentiviral (e.g. SIV) vector of the invention enables long-term transgene expression, resulting in long-term expression of a therapeutic protein. As described herein, the phrases “long-term expression”, “sustained expression”, “long-lasting expression” and “persistent expression” are used interchangeably. The retroviral/lentiviral (e.g. SIV) vectors of the present invention enable long-term transgene expression, resulting in long-term expression of a therapeutic protein, particularly by airway cells, as described herein. Long-term expression according to the present invention means expression of a therapeutic gene and/or protein, preferably at therapeutic levels, for at least 45 days, at least 60 days, at least 90 days, at least 120 days, at least 180 days, at least 250 days, at least 360 days, at least 450 days, at least 730 days or more. Preferably long-term expression means expression for at least 90 days, at least 120 days, at least 180 days, at least 250 days, at least 360 days, at least 450 days, at least 720 days or more, more preferably at least 360 days, at least 450 days, at least 720 days or more. This long-term expression may be achieved by repeated doses or by a single dose. In particular, a retroviral/lentiviral (e.g. SIV) vector of the invention may drive (increased) long- lasting expression of a therapeutic protein in an airway cell in vivo in a patient. Preferably, a retroviral/lentiviral (e.g. SIV) vector of the invention drives expression of a therapeutic protein in an airway cell for at least 45 days, more preferably at least 90 days. Repeated doses may be administered twice-daily, daily, twice-weekly, weekly, monthly, every two months, every three months, every four months, every six months, yearly, every two years, or more. Dosing may be continued for as long as required, for example, for at least six months, at least one year, two years, three years, four years, five years, ten years, fifteen years, twenty years, or more, up to for the lifetime of the patient to be treated. The retroviral/lentiviral (e.g. SIV) vectors of the invention exhibit enhanced expression of the therapeutic protein. Accordingly, the retroviral/lentiviral (e.g. SIV) vectors of the invention are capable of producing long-lasting, repeatable, high-level expression, particularly in airway cells, without inducing an undue immune response. Preferably, the invention relates to F/HN retroviral/lentiviral vectors comprising a promoter and a transgene, particularly SIV F/HN vectors. The viral vectors of the invention may be made using any suitable process known in the art. In particular, retroviral/lentiviral (e.g. SIV) vectors of the invention may be made using the methods disclosed in International Application No. PCT/GB2022/050524 which is herein incorporated by reference in its entirety. The viral vectors of the invention, particularly the retroviral/lentiviral (e.g. SIV) vectors of the invention may comprise a central polypurine tract (cPPT) and/or the Woodchuck hepatitis virus posttranscriptional regulatory elements (WPRE). An exemplary WPRE sequence is provided by SEQ ID NO: 39. A retroviral/lentiviral (e.g. SIV) vector of the invention has been modified to (i) delete the endogenous RRE as described herein; and (ii) to introduce one or more intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted. In other words, a retroviral/lentiviral (e.g. SIV) vector of the invention has a genome that has been modified to (i) delete the endogenous RRE as described herein; and (ii) to introduce one or more intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted. Any reference herein to a retroviral/lentiviral (e.g. SIV) vector of the invention which comprises an intron with an RRE inserted into the intron applies equally and without reservation to a retroviral/lentiviral (e.g. SIV) vector of the invention genome which comprises an intron with an RRE inserted into the intron. As described herein, typically an intron comprising an RRE which is introduced into a retroviral/lentiviral (e.g. SIV) vector of the invention is appropriately spliced by target cells, aiding in the maturation of a stable mRNA molecule. This results in an increased expression of the coding regions of the retroviral/lentiviral (e.g. SIV) genome, including the transgene. Thus, the introduction of an intron comprising an RRE within retroviral/lentiviral (e.g. SIV) vector results in increased expression of the transgene, which may encode a therapeutic protein. The introduction of an intron comprising an RRE into a retroviral/lentiviral (e.g. SIV) vector may increase expression of the therapeutic protein compared with expression of the therapeutic protein from a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE-comprising intron. Thus, a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron typically exhibits increased transgene expression compared with transgene expression from a corresponding retroviral/lentiviral (e.g. SIV) vector lacking said RRE-comprising intron. By way of non-limiting example, a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an AAT transgene (SERPINA1) and β- globulin/IgG chimeric intron with an inserted RRE (e.g. the RRE-comprising β-globulin/IgG chimeric intron of SEQ ID NO: 5) may increase AAT expression compared with a corresponding retroviral/lentiviral (e.g. SIV) vector which comprises the AAT transgene but which lacks the β- globulin/IgG chimeric intron with an inserted RRE (e.g. the RRE-comprising β-globulin/IgG chimeric intron of SEQ ID NO: 5). The increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron may be as defined herein. In particular, the increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron may an increase of at least about 5-fold, an increase of at least about 10-fold, an increase of at least about 50-fold, an increase of at least about 100-fold, an increase of at least about 200-fold, an increase of at least about 500-fold, an increase of at least about 600-fold or more, typically compared with expression of the therapeutic protein from a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE-comprising intron. Preferably, the increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron is at least about 10-fold, more preferably at least about 100-fold, even more preferably at least about 500-fold, typically compared with expression of the therapeutic protein from a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE- comprising intron. By way of non-limiting example, when the RRE-comprising intron is a β-globulin/IgG chimeric intron comprising an RRE, such as the β-globulin/IgG chimeric RRE-comprising intron of SEQ ID NO: 5, said intron may increase transgene expression by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising said β-globulin/IgG chimeric RRE-comprising intron by at least 600-fold, such as by about 686-fold compared with the expression of the transgene by a corresponding retroviral/lentiviral (e.g. SIV) vector without said β-globulin/IgG chimeric RRE-comprising intron. The increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron may be as defined herein. In particular, the increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron may an increase of at least about 100%, an increase of at least about 500%, an increase of at least about 1000%, an increase of at least about 5000%, an increase of at least about 10000%, an increase of at least about 20000%, an increase of at least about 50000%, an increase of at least about 60000% or more, typically compared with expression of the therapeutic protein from a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE- comprising intron. Preferably, the increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron is at least about 1000%, more preferably at least about 10000%, even more preferably at least about 50000%, typically compared with expression of the therapeutic protein from a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE-comprising intron. By way of non-limiting example, when the RRE-comprising intron is a β-globulin/IgG chimeric intron comprising an RRE, such as the β-globulin/IgG chimeric RRE-comprising intron of SEQ ID NO: 5, said intron may increase transgene expression by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising said β- globulin/IgG chimeric RRE-comprising intron by at least 60000%, such as by about 68600% compared with the expression of the transgene by a corresponding retroviral/lentiviral (e.g. SIV) vector without said β-globulin/IgG chimeric RRE-comprising intron. As described herein, typically an intron comprising an RRE which is introduced into a retroviral/lentiviral (e.g. SIV) vector of the invention is appropriately spliced by target cells, aiding in the maturation of a stable mRNA molecule. This results in an increased number of mRNA molecules (i.e. increased mRNA copy number) produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid. Thus, the introduction of an intron comprising an RRE within retroviral/lentiviral (e.g. SIV) vector results in increased expression of the transgene, which may encode a therapeutic protein. The introduction of an intron comprising an RRE into a retroviral/lentiviral (e.g. SIV) vector may increase the number of mRNA molecules produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid compared with the number of mRNA molecules produced from a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid without the RRE- comprising intron. Thus, a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE- comprising intron typically results in an increase in the number of mRNA molecules produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid compared with the number of mRNA molecules produced from a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid lacking said RRE-comprising intron. By way of non-limiting example, a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an AAT transgene (SERPINA1) and β- globulin/IgG chimeric intron with an inserted RRE (e.g. the RRE-comprising β-globulin/IgG chimeric intron of SEQ ID NO: 5) may result in an increased number of mRNA molecules produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid compared with a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid which comprises the AAT transgene but which lacks the β-globulin/IgG chimeric intron with an inserted RRE (e.g. the RRE- comprising β-globulin/IgG chimeric intron of SEQ ID NO: 5). The increase in number of mRNA molecules produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid may be as defined herein. In particular, the increase in expression of the therapeutic protein by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron may an increase of at least about 2-fold, at least about 5-fold, at least about 7-fold, at least about 10-fold, at least about 12-fold, at least about 15-fold or more, typically compared with the number of mRNA molecules produced by a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE-comprising intron. Preferably, the increase in number of mRNA molecules produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid is least about 10-fold, more preferably at least about 12-fold, even more preferably at least about 13-fold, typically compared with the number of mRNA molecules produced by a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid without the RRE- comprising intron. By way of non-limiting example, when the RRE-comprising intron is a β- globulin/IgG chimeric intron comprising an RRE, such as the β-globulin/IgG chimeric RRE-comprising intron of SEQ ID NO: 5, said intron may increase the number of mRNA molecules produced by the retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid by at least 12-fold, such as by about 13.7-fold compared with the number of mRNA molecules produced by a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid without said β-globulin/IgG chimeric RRE-comprising intron. As described herein, typically an intron comprising an RRE which is introduced into a retroviral/lentiviral (e.g. SIV) vector of the invention is appropriately spliced by target cells, aiding in the maturation of a stable mRNA molecule. This results in an increased number of mRNA molecules (i.e. increased mRNA copy number) produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid. Thus, the introduction of an intron comprising an RRE within retroviral/lentiviral (e.g. SIV) vector results in increased expression of the transgene, which may encode a therapeutic protein. The introduction of an intron comprising an RRE into a retroviral/lentiviral (e.g. SIV) vector may increase the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid compared with the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid from a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid without the RRE-comprising intron. Thus, a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron typically results in an increase in the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid compared with the number of mRNA molecules produced from a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid lacking said RRE-comprising intron. By way of non-limiting example, a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an AAT transgene (SERPINA1) and β-globulin/IgG chimeric intron with an inserted RRE (e.g. the RRE-comprising β- globulin/IgG chimeric intron of SEQ ID NO: 5) may result in an increased number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid compared with a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid which comprises the AAT transgene but which lacks the β-globulin/IgG chimeric intron with an inserted RRE (e.g. the RRE-comprising β-globulin/IgG chimeric intron of SEQ ID NO: 5). The increase in number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid may be as defined herein. In particular, the increase in number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid of the invention comprising an RRE-comprising intron may an increase of at least about 10-fold, at least about 20-fold, at least about 30-fold, at least about 40-fold, at least about 45- fold or more, typically compared with the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid from a corresponding retroviral/lentiviral (e.g. SIV) vector without the RRE-comprising intron. Preferably, the increase in number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid is least about 20-fold, more preferably at least about 30-fold, even more preferably at least about 40-fold, typically compared with the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid from a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid without the RRE-comprising intron. By way of non-limiting example, when the RRE-comprising intron is a β-globulin/IgG chimeric intron comprising an RRE, such as the β-globulin/IgG chimeric RRE-comprising intron of SEQ ID NO: 5, said intron may increase the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid by at least 40-fold, such as by about 42.2-fold compared with the number of mRNA molecules produced per copy of retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid by a corresponding retroviral/lentiviral (e.g. SIV) vector and/or vector genome plasmid without said β-globulin/IgG chimeric RRE-comprising intron. The invention also provides host cells comprising a retroviral/lentiviral (e.g. SIV) vector of the invention. Typically a host cell is a mammalian cell, particularly a human cell or cell line. Non-limiting examples of host cells include HEK293 cells (such as HEK293F or HEK293T cells) and 293T/17 cells. Commercial cell lines suitable for the production of virus are also readily available (as described herein). Rev Response Elements (RRE) The retroviral/lentiviral (e.g. SIV) vectors of the invention have been designed such that the (RNA) genome of said retroviral/lentiviral (e.g. SIV) vector comprises an intron that is not removed during manufacture of the vector, such that the final retroviral/lentiviral (e.g. SIV) vector comprises said intron, resulting in increased expression of the transgene upon transduction of target cells. To facilitate retention of the intron within the genome of the retroviral/lentiviral (e.g. SIV) vector, the endogenous RRE of the retroviral/lentiviral (e.g. SIV) vector is deleted from its position within the wildtype/unmodified retroviral/lentiviral (e.g. SIV) genome and an RRE inserted into the intron sequence. Deletion of the endogenous RRE from its position within the wildtype/unmodified retroviral/lentiviral (e.g. SIV) genome may be complete or a partial deletion, provided that if the deletion is partial, the activity of the remaining RRE sequence is decreased or completely ablated. Without being bound by theory, it is believed that partial deletion of the RRE sequence is sufficient provided that the activity of the remaining RRE sequence is insufficient for gene expression from the retroviral/lentiviral (e.g. SIV) genome, placing pressure on the retroviral/lentiviral (e.g. SIV) vector to rely on the activity of the RRE within the intron, and hence to retain the RRE inserted into the intron. Reference herein to deletion of the endogenous RRE therefore encompasses both complete and partial deletion of the endogenous RRE. Standard techniques are known in the art for the deletion of nucleic acid sequences from a nucleic acid (e.g. plasmid), and may be readily used by one of ordinary skill in the art to delete the endogenous RRE. Any RRE may be inserted into the intron to be included in the genome of the retroviral/lentiviral (e.g. SIV) vector, provided said RRE is able to facilitate retroviral/lentiviral (e.g. SIV) gene expression in the absence of the endogenous RRE in its standard position within the wildtype/unmodified retroviral/lentiviral (e.g. SIV) genome. Typically the inserted RRE is a viral RRE, particularly a retroviral RRE, even more particularly a lentiviral RRE. Standard techniques are known in the art for the insertion of nucleic acid sequences into a nucleic acid (e.g. plasmid), and may be readily used by one of ordinary skill in the art to insert an RRE into an intron according to the present invention. The RRE to be inserted into an intron may be the endogenous RRE of the retroviral/lentiviral (e.g. SIV) vector. Thus, the endogenous RRE may be deleted from within the wildtype/unmodified retroviral/lentiviral (e.g. SIV) genome and inserted into an intron which is itself introduced into a retroviral/lentiviral (e.g. SIV) vector of the invention. In other words, the endogenous RRE is moved from its position within the wildtype/unmodified retroviral/lentiviral (e.g. SIV) genome, and inserted into an intron. By way of non-limiting example, in an SIV vector of the invention, the endogenous SIV RRE has been deleted from within the wildtype/unmodified SIV genome, and an intron into which the endogenous SIV RRE has been inserted is itself introduced into the SIV vector. The RRE to be inserted into an intron may be an exogenous RRE. In the case of a retroviral vector of the invention, an exogenous RRE may be an RRE from a different retrovirus. In the case of a lentiviral vector of the invention, an exogenous RRE may be an RRE from a different lentivirus. By way of non-limiting example, an HIV vector of the invention may have its endogenous HIV RRE deleted and an intron comprising a SIV RRE introduced into the HIV genome. Preferably the RRE sequence inserted into an intron within a retroviral/lentiviral (e.g. SIV) vector of the invention is the same as the endogenous RRE sequence which is deleted from the retroviral/lentiviral (e.g. SIV) genome. Thus, preferably, the RRE is from the same virus as the viral vector, but is inserted into the (chimeric) intron, rather than being present in its standard location within the viral genome. By way of non-limiting example, the viral vector may be an SIV vector in which the SIV RRE has been deleted from the genome and a (chimeric) intron introduced into which an SIV RRE has been inserted. By way of further non-limiting example, the viral vector may be an HIV vector, and the RRE may be an HIV RRE, but the HIV RRE is inserted within a (chimeric) intron, rather than in the standard location of the HIV RRE within the HIV genome. The RRE to be inserted into the intron may be less than 1,000 bp, such as less than 900 bp or less than 800 bp may be preferred. Without being bound by theory, it is believed that smaller RRE- comprising introns may be more suitable for general applicability, allowing for greater flexibility in terms of the additional elements to be included within the retroviral/lentiviral (e.g. SIV) genome. Particularly preferred are RRE of between about 750 bp to about 800 bp, such as about 760 bp, such as the exemplified SIV RRE of the invention. The RRE to be inserted into the intron may be an SIV RRE. The SIV RRE may comprise or consist of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 1. Preferably, the SIV RRE comprises or consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 1. More preferably, the SIV RRE consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 1. Still more preferably, the SIV RRE comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 1. The RRE to be inserted into the intron may be an HIV RRE. The HIV RRE may comprise or consist of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 50. Preferably, the HIV RRE comprises or consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 50. More preferably, the HIV RRE consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 50. Still more preferably, the HIV RRE comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 50. According to the invention, typically the RRE inserted into the intron does not form part of the mature mRNA expressed within the host/target cells, as the RRE will be spliced out as part of the intron. Introns A retroviral/lentiviral (e.g. SIV) vector of the invention has been modified to (i) delete the endogenous RRE as described herein; and (ii) to introduce one or more intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise one or more intron, such as one, two, three, four, five or more introns. Typically a retroviral/lentiviral (e.g. SIV) vector of the invention comprises one or two introns, preferably one intron. Wildtype/unmodified retroviral/lentiviral (e.g. SIV) vector genomes do not comprise introns. Therefore any and all references herein to retroviral/lentiviral (e.g. SIV) vectors/ vector genomes comprising one or more introns refers to retroviral/lentiviral (e.g. SIV) vectors/ vector genomes into which one or more introns have been introduced. In other words, any intron comprised in a retroviral/lentiviral (e.g. SIV) vector/vector genome of the invention is an intron which has been introduced into said retroviral/lentiviral (e.g. SIV) vector/vector genome, as described herein. The size of the one or more intron to be inserted is not particularly limited, provided that the retroviral/lentiviral (e.g. SIV) vector/vector genome comprising the one or more intron is within the packing limit for the retroviral/lentiviral (e.g. SIV) vector/vector genome. As the retroviral/lentiviral (e.g. SIV) vector/vector genome will comprise other elements (including the genome backbone, transgene and transgene promoter) in addition to the intron, the upper size limit of the one or more intron to be inserted may be calculated as: UL_i = VPL – GE where UL_i is the upper size limit of intron, VPL is the retroviral/lentiviral (e.g. SIV) packing limit, and GE is the total size of other retroviral/lentiviral (e.g. SIV) genome elements. Retroviral/lentiviral (e.g. SIV) vectors typically have a packing limit of approximately 10 kb. Therefore, typically the size of the one or more intron to be inserted is less than about 5,000 bp, accounting for the other elements that must be present within the retroviral/lentiviral (e.g. SIV) genome. Introns of less than 1,000 bp, such as less than 900 bp or less than 800 bp may be preferred. Without being bound by theory, it is believed that smaller introns of this type may be more suitable for general applicability, allowing for greater flexibility in terms of the additional elements to be included within the retroviral/lentiviral (e.g. SIV) genome. Particularly preferred are introns of between about 750 bp to about 800 bp, such as about 770 bp, such as the exemplified β-globulin/IgG chimeric intron of the invention. The one or more intron may be introduced at any position within the retroviral/lentiviral (e.g. SIV) genome. Typically the one or more intron is introduced at a position within the retroviral/lentiviral (e.g. SIV) genome that does not disrupt the function of the retroviral/lentiviral (e.g. SIV) genome or any part thereof. By way of non-limiting example, the one or more intron may be inserted at any position within the retroviral/lentiviral (e.g. SIV) genome, provided that translation of the transgene is not decreased. Preferably the intron is introduced between a transgene and the promoter operably linked to said transgene. Thus, preferably a retroviral/lentiviral (e.g. SIV) vector/genome of the invention comprises an intron between the transgene and the promoter operably linked to said transgene. According to the invention, the RRE-comprising intron does not comprise the transgene to be expressed. The sequence of the intron to be introduced is not particularly limited. Indeed, as exemplified herein, the present inventors have shown that the retroviral RRE remains functional in different contexts, and thus that RRE function does not depend on the specific intron sequence. Further, it has been known the art for decades that non-chimeric introns can be divided and combined with other DNA sequences (see for example, Choi, T. et al. (1990) Molecular and Cellular Biology, 11:6, 3070- 3074, which is herein incorporated by reference). Thus, any appropriate intron may be used according to the present invention. In other words, any intron may have an RRE sequence inserted, and said intron/RRE introduced into a retroviral/lentiviral (e.g. SIV) vector/genome of the invention. The intron may be a naturally occurring, recombinant, or artificial, such as a chimeric intron. The intron may be a viral intron. Non-limiting examples of introns include: SV40 intron, Ef1-α intron 1, CMB intron A and adenovirus tripartite leader sequence intron. The intron may be a chimeric intron or a non-chimeric intron. Preferably the intron is a chimeric intron, with the chimeric β-globulin/IgG intron exemplified herein being particularly preferred. Typically the intron is a chimeric intron, such that retroviral/lentiviral (e.g. SIV) vectors of the invention comprise a chimeric intron. A chimeric intron is an artificial intron which comprises or consists of sequences from two or more different introns. Non-limiting examples of chimeric introns include a β-globulin/IgG chimeric intron and the chimeric intron from the CAGGS promoter. The latter comprises the splice donor from chicken β-actin and the splice acceptor from rabbit β-globulin. Preferably, a chimeric intron according to the present invention is a β-globulin/IgG chimeric intron, such as that exemplified herein. Particularly preferred is a β-globulin/IgG chimeric intron which comprises or consists of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 4. Preferably, the β-globulin/IgG chimeric intron comprises or consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 4. More preferably, the β-globulin/IgG chimeric intron consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 4. Still more preferably, the β-globulin/IgG chimeric intron comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 4. A chimeric intron from the CAGGS promoter may comprise or consist of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 48. Preferably, the chimeric intron from the CAGGS promoter comprises or consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 48. More preferably, the chimeric intron from the CAGGS promoter consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 48. Still more preferably, the chimeric intron from the CAGGS promoter comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 48. The RRE is introduced at a position within the intron which does not disrupt the splice donor and/or acceptor sites of the intron, to allow for the intron to be spliced correctly within a target cell. The splice donor and/or acceptor sites of a particular intron may be readily determined using routine methods and techniques, for example as described in Desmet et al. (Nucleic Acids Res. 2009 May; 37(9): e67) and Baten et al. (BMC Bioinformatics volume 7, Article number: S15 (2006)), both of which are herein incorporated by reference in their entirety. In particular, the inventors have found that placing an RRE within 200 bp 5' of the splice acceptor’s branch site (and polypyrimidine tract), such as within 100 bp 5' of the splice acceptor’s branch site, within 50 bp 5' of the splice acceptor’s branch site or within 20 bp 5' of the splice acceptor’s branch site, is advantageous as it allows the RRE to be inserted whilst allowing efficient splicing of the intron to produce retroviral/lentiviral (e.g. SIV) mRNA. In particular, and as exemplified herein, an RRE may preferably be inserted within 20 bp 5' of the splice acceptor’s branch site (and polypyrimidine tract), such 20 bp 5' of the splice acceptor’s branch site, 19 bp 5’ of the splice acceptor’s branch site, 18 bp 5' of the splice acceptor’s branch site or less, with 18 bp 5' of the splice acceptor’s branch site being particularly preferred. Typically at least there is a space of at least 5 bp, at least 10 bp or at least 15 bp between the RRE insertion site and the splice acceptor’s branch site, to ensure that this branch site is not disrupted by RRE insertion. Thus, an RRE may be inserted at a location between about 5-20 bp 5' of the splice acceptor’s branch site (and polypyrimidine tract), such as between about 10-20 bp 5' of the splice acceptor’s branch site or between about 15-20 bp 5' of the splice acceptor’s branch site, with RRE insertion 18 bp 5' of the splice acceptor’s branch site being preferred. The RRE insertion site devised by the present inventors differs from insertion sites attempted in the art, which are typically closer to the splice donor site than the splice acceptor site. The RRE insertion site of the invention is also typically designed such that any other regulatory sequences within the intron are not disrupted by RRE insertion. When the intron is a chimeric intron, the RRE may be inserted at a junction between the sequences from the two or more different introns. By way of non-limiting example, where the chimeric intron is a β-globulin/IgG chimeric intron, the RRE may be inserted at the junction between the β-globulin intron sequence (the 5’ portion of the chimeric β-globulin/IgG chimeric intron) and the IgG intron sequence (the 3’ portion of the β-globulin/IgG chimeric intron). In preferred embodiments relating to the use of a β-globulin/IgG chimeric intron, the RRE may be inserted between a splice donor site and a splice acceptor site, wherein (a) the splice donor site comprises or consists of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 2 (from β-globulin); and/or (b) the splice acceptor site comprises or consists of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 3 (from IgG). Preferably (a) the splice donor site comprises or consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 2; and/or (b) the splice acceptor site comprises or consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 3. More preferably (a) the splice donor site consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 2; and/or (b) the splice acceptor site consists of a nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 3. Still more preferably (a) the splice donor site comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 2; and/or (b) the splice acceptor site comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 3. The intron may be introduced into the retroviral/lentiviral (e.g. SIV) vector/genome in either forward or reverse orientation. Preferably, the intron is introduced into the retroviral/lentiviral (e.g. SIV) vector/genome in the forward orientation. Preferably, and as exemplified herein, the intron is a β-globin/IgG chimeric intron and the RRE is an SIV RRE. The β-globin/IgG chimeric intron comprising a SIV RRE may comprise or consist of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 5. Preferably, the β- globin/IgG chimeric intron comprising a SIV RRE comprises or consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 5. More preferably, the β- globin/IgG chimeric intron comprising a SIV RRE consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 5. Still more preferably, the β-globin/IgG chimeric intron comprising a SIV RRE comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 5. An intron into which an RRE has been inserted according to the present invention may be referred to interchangeably herein as an “RRE-comprising intron”, an “intron comprising an RRE”, “an intron with an inserted RRE”, and an “intron with an introduced RRE”. Typically the intron comprising an RRE within a retroviral/lentiviral (e.g. SIV) vector/genome is appropriately spliced by target cells, aiding in the maturation of a stable mRNA molecule. This results in an increased expression of the coding regions of the retroviral/lentiviral (e.g. SIV) genome, including the transgene. Thus, the introduction of an intron comprising an RRE within retroviral/lentiviral (e.g. SIV) vector/genome results in increased expression of the transgene, which may encode a therapeutic protein. When inserting an RRE into an intron according to the present invention, it is important that the position at which the RRE is inserted into the intron is carefully defined and controlled. This is because it is necessary to insert the RRE at such a position that the RRE is still able to function, and the intron is still recognised as such during transcription of the retroviral/lentiviral (e.g. SIV) genome. Provided that the position at which the RRE is inserted into the intron is carefully defined and controlled as described herein, then the precise sequence of the intron is not limiting. Accordingly, also provided is a method of designing and/or producing an RRE-comprising intron according to the invention. Said method may comprise or consist of the steps of (a) identifying the splice donor and splice acceptor sequences within the intron; and (b) inserting the RRE into the intron such that the splice donor and split acceptor sequences remain intact. The method may further comprise one or more steps to delete the endogenous RRE from the retroviral/lentiviral (e.g. SIV) genome. Standard techniques insert and/or delete nucleic acid sequences from a nucleic acid (e.g. plasmid) are known in the art, and may be used by one of ordinary skill to insert the RRE-comprising intron and/or to delete the endogenous RRE according to the invention. Transgenes and Promoters A retroviral/lentiviral (e.g. SIV) vector the invention typically comprises a transgene encoding for a therapeutic protein. A therapeutic protein is one which has potential utility in the treatment or prevention of a disease or condition, such as those describe herein. Thus, a retroviral/lentiviral (e.g. SIV) vector of the invention comprises a transgene encoding a protein which has a therapeutic effect on a disease or condition to be treated. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a transgene encoding a therapeutic protein which is a functional or wild-type form of a protein which is present in a patient to be treated in a dysfunctional form (whether the dysfunction is inherent or acquired). As used herein, the phrase "inherent dysfunction" refers to a protein which is innately dysfunctional due to genetic factors and the phrase "acquired dysfunction" refers to a protein which is dysfunctional due to environmental or other factors after birth. By way of non-limiting example, CFTR is an example of a protein which is inherently dysfunctional in patients with cystic fibrosis. Thus, a retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a transgene encoding a therapeutic protein which is a functional or wild-type form of a protein which is present in a patient, but which that has become dysfunctional due to a genetic disease, such as a genetic respiratory disease. The retroviral/lentiviral (e.g. SIV) vectors of the present invention are useful in the treatment of diseases via their use in expressing therapeutic proteins in target cells, wherein the therapeutic protein exerts its therapeutic effects: (i) within the target cells; (ii) by secretion from said cells into the surrounding tissue; or (iii) for secretion from said cells into the circulatory system. A retroviral/lentiviral (e.g. SIV) vector of the present invention may be pseudotyped to target airway cells of the respiratory tract (e.g. by pseudotyping with F and HN proteins from a respiratory paramyxovirus such as a Sendai virus), as described herein. Such retroviral/lentiviral (e.g. SIV) vectors of the invention are useful in the treatment of diseases via their use in expressing therapeutic proteins in airway cells: (i) within the respiratory tract; (ii) for secretion from said cells into the lumen of the respiratory tract; and (iii) for secretion from said cells into the circulatory system. The therapeutic protein may be selected from: (a) a secreted therapeutic protein, optionally alpha-1-antitrypsin (AAT), Factor VIII, Surfactant Protein B (SFTPB), Factor VII, Factor IX, Factor X, Factor XI, von Willebrand Factor, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), Surfactant Protein C (SP-C), an anti-inflammatory protein (e.g. IL-10 or TGGβ) or monoclonal antibody, an anti-inflammatory decoy and a monoclonal antibody against an infectious agent; or (b) CFTR, CSF2RA, CSF2RB and ATP-binding cassette sub-family member A (ABCA3). Preferred examples of therapeutic proteins include AAT, GM-CSF, FVIII, CFTR, decorin, TRIM72 and ABCA3. The transgene may encode: (i) a therapeutic protein that is secreted into epithelial lining fluid and/or blood); (ii) a therapeutic protein that is secreted into blood); or (iii) a therapeutic membrane protein). Preferred examples of these classes of transgenes include (i) AAT; (ii) FVIII; and (iii) CFTR. In some embodiments, the therapeutic protein is not an antibody, particularly not a monoclonal antibody and/or not a β-globulin gene. In such embodiments, the therapeutic protein may be selected from: (a) a secreted therapeutic protein, optionally alpha-1-Antitrypsin (AAT), Factor VIII, Surfactant Protein B (SFTPB), Factor VII, Factor IX, Factor X, Factor XI, von Willebrand Factor, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), Surfactant Protein C (SP-C), an anti- inflammatory protein (e.g. IL-10, TGGβ, or TNF-alpha) and an anti-inflammatory decoy; or (b) CFTR, CSF2RA, CSF2RB and ATP-binding cassette sub-family member A (ABCA3). The retroviral/lentiviral (e.g. SIV) vectors of the invention are particularly efficient at driving the expression, secretion and/or membrane insertion of proteins (e.g. therapeutic proteins as described herein) by airway cells. This is particularly the case when the retroviral/lentiviral (e.g. SIV) vectors are F/HN pseudotyped viral vectors of the invention (as described herein), which are efficient at targeting cells in the airway epithelium. As such, for therapeutic applications the retroviral/lentiviral (e.g. SIV) vectors of the invention are typically delivered to cells of the respiratory tract, including the cells of the airway epithelium. In other words, the retroviral/lentiviral (e.g. SIV) vectors of the invention are typically delivered to airway cells as described herein. Accordingly, the retroviral/lentiviral (e.g. SIV) vectors of the invention are particularly suited for treatment of diseases or disorders of the airways, respiratory tract, or lung. Typically, the retroviral/lentiviral (e.g. SIV) vectors of the invention may be used for the treatment of a genetic respiratory disease. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a transgene encoding a polypeptide or protein that is therapeutic for the treatment of such diseases, particularly a disease or disorder of the airways, respiratory tract, or lung. The transgene and therapeutic protein of the invention are not limited, one of ordinary skill in the art will be able to identify therapeutic proteins which may be usefully delivered according to the invention, particularly in the context of genetic diseases, particularly genetic respiratory diseases and diseases or disorders of the airways, respiratory tract, or lung such as those described herein. Accordingly, a retroviral/lentiviral (e.g. SIV) vectors of the invention may comprise a nucleic acid sequence encoding a therapeutic protein selected from: (a) a secreted therapeutic protein, optionally alpha-1-antitrypsin (AAT), Factor VIII, Surfactant Protein B (SFTPB), ADAMTS13, Factor VII, Factor IX, Factor X, Factor XI, von Willebrand Factor, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), Surfactant Protein C (SP-C), an anti-inflammatory protein (e.g. IL-10, TGGβ, or TNF- alpha) or monoclonal antibody, an anti-inflammatory decoy and a monoclonal antibody against an infectious agent; or (b) CFTR, CSF2RA, CSF2RB, ATP-binding cassette sub-family member A (ABCA3), DNAH5, DNAH11, DNAI1, and DNAI2. Other examples of therapeutic proteins that may be encoded by a transgene comprised in a retroviral/lentiviral (e.g. SIV) vector of the invention include genes related to or associated with other surfactant deficiencies. Preferred examples of therapeutic proteins include AAT, GM-CSF, FVIII, CFTR, ADAMTS13, SFTPB, decorin, TRIM72 and ABCA3. The therapeutic protein encoded by a retroviral/lentiviral (e.g. SIV) vector of the invention may be an AAT. An example of an AAT therapeutic transgene (SERPINA1) is provided by SEQ ID NO: 6, or by the complementary sequence of SEQ ID NO: 7. SEQ ID NO: 6 is a codon-optimized CpG depleted AAT transgene (SERPINA1) previously designed by the present inventors to enhance translation in human cells. Such optimisation has been shown to enhance gene expression by up to 15-fold. Variants of same sequence (as defined herein) which possess the same technical effect of enhancing translation compared with the unmodified (wild-type) AAT gene sequence are also encompassed by the present invention. The therapeutic protein encoded by said AAT transgene, may be exemplified by the polypeptide of SEQ ID NO: 8. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to any one of SEQ ID NO: 6, 7 or 8. The therapeutic protein encoded by a retroviral/lentiviral (e.g. SIV) vector of the invention may be an FVIII. Examples of a FVIII therapeutic transgene are provided by SEQ ID NOs: 9 and 10, or by the respective complementary sequences of SEQ ID NO: 11 and 12. The polypeptide encoded by the FVIII transgene, may be exemplified by the polypeptide of SEQ ID NO: 13 or 14. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to any one of SEQ ID NOs: 9 to 14. Preferably, the therapeutic protein encoded by a retroviral/lentiviral (e.g. SIV) vector of the invention is a CFTR. An example of a CFTR transgene is provided by SEQ ID NO: 15. The polypeptide encoded by said CFTR transgene, may be exemplified by the polypeptide of SEQ ID NO: 16. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to SEQ ID NO: 15 or 16. The therapeutic protein encoded by a retroviral/lentiviral (e.g. SIV) vector of the invention may be GM-CSF. A GM-CSF transgene may comprise or consist of SEQ ID NO: 17 (human). The polypeptide encoded by the GM-CSF transgene may be exemplified by the polypeptide of SQE ID NO: 18 (human). Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to any one of SEQ ID NOs: 17 and 18. The transgene may encode decorin. An example of a DCN transgene is provided by SEQ ID NO: 21. The polypeptide encoded by said DCN transgene, may be exemplified by the polypeptide of SEQ ID NO: 22. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to SEQ ID NO: 21 or 22. The transgene may encode TRIM72. An example of a TRIM72 transgene is provided by SEQ ID NO: 23. The polypeptide encoded by said TRIM72 transgene, may be exemplified by the polypeptide of SEQ ID NO: 24. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to SEQ ID NO: 23 or 24. The transgene may encode ABCA3. An example of a ABCA3 transgene is provided by SEQ ID NO: 25. The polypeptide encoded by said ABCA3 transgene, may be exemplified by the polypeptide of SEQ ID NO: 26. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to SEQ ID NO: 25 or 26. The transgene may encode SFTPB. An example of a SFTPB transgene is provided by SEQ ID NO: 40. The polypeptide encoded by said SFTPB transgene, may be exemplified by the polypeptide of SEQ ID NO: 41. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to SEQ ID NO: 40 or 41. The transgene may encode ADAMTS13. An example of a ADAMTS13 transgene is provided by SEQ ID NO: 42. The polypeptide encoded by said ADAMTS13 transgene, may be exemplified by the polypeptide of SEQ ID NO: 43. Variants thereof (as described therein) are also included, particularly variants with at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) to SEQ ID NO: 42 or 43. The therapeutic protein encoded by a retroviral/lentiviral (e.g. SIV) vector of the invention may be encoded by any one of SFTPB, SFTPC, ADAMTS13, Factor V, Factor VII, Factor IX, Factor X and/or Factor XI, von Willebrand Factor, GM-CSF, ABCA3, TRIM72 or DCN, or other known related gene. When the respiratory tract epithelium is targeted for delivery of the retroviral/lentiviral (e.g. SIV) vectors of the invention, the therapeutic protein may be AAT, SFTPB, or GM-CSF. The therapeutic protein may be a monoclonal antibody (mAb) against an infectious agent (bacterial, fungal or viral, e.g. the SARS-Co-V2 virus). The therapeutic protein may be anti-TNF alpha. The therapeutic protein may be one implicated in an inflammatory, immune or metabolic condition. A retroviral/lentiviral (e.g. SIV) vector of the invention may be delivered to the cells of the respiratory tract to allow production of proteins to be secreted into circulatory system. In such embodiments, the therapeutic protein may be any one of Factor VII, Factor VIII, Factor IX, Factor X, Factor XI and/or von Willebrand’s factor. Such a retroviral/lentiviral (e.g. SIV) vector of the invention may be used in the treatment of diseases, particularly cardiovascular diseases and blood disorders, preferably blood clotting deficiencies such as haemophilia. Again, the therapeutic protein may be an mAb against an infectious agent or a protein implicated in an inflammatory, immune or metabolic condition, such as, lysosomal storage disease. The retroviral/lentiviral (e.g. SIV) vector comprises a promoter operably linked to a transgene, enabling expression of the transgene. Typically the promoter is a hybrid human CMV enhancer/EF1a (hCEF) promoter. This hCEF promoter may lack the intron corresponding to nucleotides 570-709 and the exon corresponding to nucleotides 728-733 of the hCEF promoter. A preferred example of an hCEF promoter sequence of the invention is provided by SEQ ID NO: 27. The promoter may be a CMV promoter. An example of a CMV promoter sequence is provided by SEQ ID NO: 28. The promoter may be a human elongation factor 1a (EF1a) promoter. An example of a EF1a promoter is provided by SEQ ID NO: 29. Other promoters for transgene expression are known in the art and their suitability for the retroviral/lentiviral (e.g. SIV) vectors of the invention determined using routine techniques known in the art. Non-limiting examples of other promoters include UBC and UCOE. As described herein, the promoter may be modified to further regulate expression of the transgene of the invention. The promoter included in the retroviral/lentiviral (e.g. SIV) vector of the invention may be specifically selected and/or modified to further refine regulation of expression of the therapeutic gene. Again, suitable promoters and standard techniques for their modification are known in the art. As a non-limiting example, a number of suitable (CpG-free) promoters suitable for use in the present invention are described in Pringle et al. (J. Mol. Med. Berl. 2012, 90(12): 1487-96), which is herein incorporated by reference in its entirety. Preferably, the retroviral/lentiviral vectors (particularly SIV F/HN vectors) of the invention comprise a hCEF promoter having low or no CpG dinucleotide content. The hCEF promoter may have all CG dinucleotides replaced with any one of AG, TG or GT. Thus, the hCEF promoter may be CpG-free. A preferred example of a CpG-free hCEF promoter sequence of the invention is provided by SEQ ID NO: 27. The absence of CpG dinucleotides typically further improves the performance of retroviral/lentiviral (e.g. SIV) vectors of the invention and in particular in situations where it is not desired to induce an immune response against an expressed antigen or an inflammatory response against the delivered expression construct. The elimination of CpG dinucleotides reduces the occurrence of flu-like symptoms and inflammation which may result from administration of constructs, particularly when administered to the airways. The retroviral/lentiviral (e.g. SIV) vector of the invention may be modified to allow shut down of gene expression. Standard techniques for modifying the vector in this way are known in the art. As a non-limiting example, Tet-responsive promoters are widely used. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF promoter and a CFTR transgene, including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF promoter and an AAT transgene (SERPINA1), including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF or CMV promoter and an FVIII transgene, including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF or CMV promoter and an DCN transgene, including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF or CMV promoter and an TRIM72 transgene, including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF or CMV promoter and an ABCA3 transgene, including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF or CMV promoter and an SFTPB transgene, including those described herein. A retroviral/lentiviral (e.g. SIV) vector of the invention may comprise a hCEF or CMV promoter and an ADAMTS13 transgene, including those described herein. The retroviral/lentiviral (e.g. SIV) vector of the invention comprises a nucleic acid encoding a therapeutic protein (said nucleic acid is referred to interchangeably herein as a transgene). The nucleic acid sequence encodes a gene product, e.g., a protein, particularly a therapeutic protein. For example, the retroviral/lentiviral (e.g. SIV) vector may comprise a transgene encoding an AAT, GM-CSF, FVIII, SFTPB, ADAMTS13, CFTR, decorin, TRIM72 or ABCA3 and said transgene comprises (or consists of) a nucleic acid sequence having at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99 or 100%) sequence identity to the AAT, GM-CSF, FVIII, SFTPB, ADAMTS13, CFTR, decorin, TRIM72 or ABCA3 transgene respectively, examples of which are described herein. In a further embodiment, the transgene encoding AAT, GM-CSF, FVIII, SFTPB, ADAMTS13, CFTR, decorin, TRIM72 or ABCA3 comprises (or consists of) a nucleic acid sequence having at least 95% (such as at least 95, 96, 97, 98, 99 or 100%) sequence identity to the AAT, GM-CSF, FVIII, SFTPB, ADAMTS13, CFTR, decorin, TRIM72 or ABCA3 nucleic acid sequence respectively, examples of which are described herein. The nucleic acid sequence encoding CFTR may be provided by SEQ ID NO: 15, the nucleic acid sequence encoding AAT may be provided by SEQ ID NO: 6, or by the complementary sequence of SEQ ID NO: 7 and/or the nucleic acid sequence encoding FVIII may be provided by SEQ ID NO: 11 or 12, or by the respective complementary sequences of SEQ ID NO: 13 or 14, and/or the nucleic acid sequence encoding SFTPB may be provided by SEQ ID NO: 40, and/or the nucleic acid sequence encoding ADAMTS13 may be provided by SEQ ID NO: 42, and/or the nucleic acid sequence encoding GM-CSF may be provided by SEQ ID NO: 17, the nucleic acid sequence encoding decorin may be provided by SEQ ID NO: 21, the nucleic acid sequence encoding TRIM72 may be provided by SEQ ID NO: 23, and/or the nucleic acid sequence encoding ABCA3 may be provided by SEQ ID NO: 25, or variants thereof. The amino acid sequence of the therapeutic protein may be a functional variant having at least 95% (such as at least 95, 96, 97, 98, 99 or 100%) sequence identity to the functional protein. For example, an AAT, FVIII, SFTPB, ADAMTS13, CFTR, GM-CSF, decorin, TRIM72 and/or ABCA3 polypeptide encoded by the respective AAT, FVIII, SFTPB, ADAMTS13, CFTR, CSF2, DCN, TRIM72, and/or ABCA3 transgene may comprise (or consist of) an amino acid sequence having at least 95% (such as at least 95, 96, 97, 98, 99 or 100%) sequence identity to the functional AAT, FVIII, SFTPB, ADAMTS13, CFTR, GM-CSF, decorin, TRIM72 and/or ABCA3 polypeptide sequence respectively. The transgene encoding for a therapeutic protein may include a nucleic acid sequence encoding for the endogenous signal peptide of the therapeutic protein, or may exclude a nucleic acid sequence encoding for this signal peptide. All disclosure herein relates to both transgenes and therapeutic proteins including and excluding endogenous signal peptides unless explicitly stated. By way of non-limiting example, sequence identity of variants, and/or lengths of fragments may be based on the sequence with or without a signal peptide. A retroviral/lentiviral (e.g. SIV) vector of the invention typically further comprises a Rev protein. This Rev protein is typically provided by (encoded by) one of the plasmids used in the manufacture of the retroviral/lentiviral (e.g. SIV) vector, as described herein. By way of non-limiting example, the Rev protein may be provided by the Rev plasmid (pDNA2b), wherein separate plasmids are used to provide the Gag-Pol and Rev proteins, or the Rev protein may be provided by the Rev-Gag- Pol plasmid, when a single plasmid is used to provide the Gag-Pol and Rev proteins. An exemplary pDNA2b plasmid, as described herein is pGM299, as shown in Figure 2D and with a sequence represented by SEQ ID NO: 33. An exemplary Rev protein is the rSIV Rev protein which comprises or consists of the amino acid sequence of SEQ ID NO: 44. This Rev protein is encoded by the pGM299 plasmid. Nucleic Acids The present invention also provides a nucleic acid comprising or consisting of an intron (e.g. a chimeric intron) into which an RRE has been introduced, as described herein. Any intron as described herein may be comprised in a nucleic acid of the invention. Similarly, any RRE as described herein may be comprised in a nucleic acid of the invention. The intron and RRE may each be selected independently, e.g. from those described herein. Any and all disclosure herein in relation to introns and/or RRE in the context of retroviral/lentiviral (e.g. SIV) vectors of the invention applies equally and without reservation to introns and/or RRE in the context of nucleic acids of the invention. By way of non-limiting example, a nucleic acid of the invention may comprise any intron as described herein, for example a β-globin/IgG chimeric intron comprising or consisting of SEQ ID NO: 4, or a variant thereof, as described herein. Alternatively or additionally, by way of further non-limiting example, a nucleic acid of the invention may comprise any RRE as described herein, for example a SIV RRE comprising or consisting of SEQ ID NO: 1, or a variant thereof, as described herein. Preferably, and as exemplified herein, a nucleic acid of the invention comprises or consists of a β-globin/IgG chimeric intron and an SIV RRE. Accordingly, a nucleic acid of the invention may comprise or consist of a β-globin/IgG chimeric intron comprising a SIV RRE which comprise or consist of a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or more, up to 100% sequence identity to SEQ ID NO: 5. Preferably, a nucleic acid of the invention may comprise or consist of a β-globin/IgG chimeric intron comprising a SIV RRE which comprises or consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 5. More preferably, a nucleic acid of the invention may comprise or consist of a β-globin/IgG chimeric intron comprising a SIV RRE which consists of nucleic acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 5. Still more preferably, a nucleic acid of the invention may comprise or consist of a β-globin/IgG chimeric intron comprising a SIV RRE which comprises or consists, particularly consists of, the nucleic acid sequence of SEQ ID NO: 5. A nucleic acid of the invention may further comprise a transgene, typically which encodes a therapeutic protein as described herein. Any and all disclosure herein in relation to transgenes in the context of retroviral/lentiviral (e.g. SIV) vectors of the invention applies equally and without reservation to transgenes in the context of nucleic acids of the invention. In some preferred embodiments, the therapeutic protein encoded by the transgene is AAT or CFTR. A nucleic acid of the invention comprising an RRE-comprising intron may exhibit increased expression of the transgene compared with a corresponding nucleic acid lacking said intron. The disclosure herein relating to the increase in transgene expression by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising an RRE-comprising intron applies equally and without reservation to the increase in transgene expression exhibited by a nucleic acid of the invention. By way of non-limiting example, the increase in expression of the therapeutic protein by a nucleic acid of the invention comprising an RRE-comprising intron may an increase of at least about 5- fold, an increase of at least about 10-fold, an increase of at least about 50-fold, an increase of at least about 100-fold, an increase of at least about 200-fold, an increase of at least about 500-fold, an increase of at least about 600-fold or more, typically compared with expression of the therapeutic protein from a corresponding nucleic acid without the RRE-comprising intron. A nucleic acid of the invention enables long-term transgene expression, resulting in long-term expression of a therapeutic protein. As described herein, the phrases “long-term expression”, “sustained expression”, “long-lasting expression” and “persistent expression” are used interchangeably. Long-term expression according to the present invention means expression of a therapeutic protein, preferably at therapeutic levels, for at least 45 days, at least 60 days, at least 90 days, at least 120 days, at least 180 days, at least 250 days, at least 360 days, at least 450 days, at least 730 days or more. Preferably long-term expression means expression for at least 90 days, at least 120 days, at least 180 days, at least 250 days, at least 360 days, at least 450 days, at least 720 days or more, more preferably at least 360 days, at least 450 days, at least 720 days or more. In particular, a nucleic acid of the invention may drive (increased) long-lasting expression of a therapeutic protein in an airway cell in vivo in a patient. Preferably, a nucleic acid of the invention drives expression of a therapeutic protein in an airway cell for at least 45 days, more preferably at least 90 days. The nucleic acid of the nucleic acid may be as defined herein. The nucleic acid may comprise DNA and/or RNA. Preferably the nucleic acid is DNA. A nucleic acid of the invention may optionally be codon optimised for expression in a particular cell type, for example, eukaryotic cells (e.g. mammalian cells, yeast cells, insect cells or plants cells) or prokaryotic cells (e.g. E.coli). The term “codon optimised” refers to the replacement of at least one codon within a base polynucleotide sequence with a codon that is preferentially used by the host organism in which the polynucleotide is to be expressed. Typically, the most frequently used codons in the host organism are used in the codon-optimised polynucleotide sequence. Methods of codon optimisation are well known in the art. It will be understood by a skilled person that numerous different polynucleotides can encode the same polypeptide as a result of the degeneracy of the genetic code. It is also understood that skilled persons may, using routine techniques, make nucleotide substitutions that do not affect the polypeptide sequence encoded by the nucleic acid molecules to reflect the codon usage of any particular host organism in which the polypeptides are to be expressed. Therefore, unless otherwise specified, a nucleic acid that encodes a therapeutic protein of the invention includes all polynucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. A nucleic acid cassette of the invention typically comprises a promoter operably linked to the nucleic acid sequence encoding the therapeutic protein. By operably linked, it is meant that the promoter is configured to express the nucleic acid sequence encoding the signal peptide and/or the nucleic acid sequence encoding the therapeutic protein. The disclosure herein in relation to promoters in the context of retroviral/lentiviral (e.g. SIV) vectors of the invention applies equally to nucleic acids of the invention. In some preferred embodiments, the promoter is a hCEF promoter as described herein. The nucleic acids of the invention may include at least one part of a vector, in particular, regulatory elements. By way of non-limiting example, the promoter (e.g. the hCEFI promoter) within a nucleic acid cassette of the invention may be used to express more than one polypeptide, including one or more therapeutic proteins. Thus, the nucleic acid may comprise a nucleic acid sequence which, when transcribed, gives rise to multiple polypeptides, for instance a transcript may contain multiple open reading frames (ORFs) and also one or more Internal Ribosome Entry Sites (IRES) to allow translation of ORFs after the first ORF. A transcript may be polycistronic, i.e. it may be translated to give a polypeptide which is subsequently cleaved to give a plurality of polypeptides. Alternatively, a nucleic acid of the invention may comprise multiple promoters and hence give rise to a plurality of transcripts and hence a plurality of polypeptides, including a plurality of therapeutic proteins. Nucleic acids may, for instance, express one, two, three, four or more polypeptides via a promoter (e.g. hCEFI) or promoters. A nucleic acid may comprise one or more translation initiation sequences (TIS). Translation initiation plays an important role in mRNA translation, canonically a methionyl tRNA unique for initiation (Met-tRNAi) identifies the AUG start codon and triggers the downstream translation process. Non-canonical start codons (e.g. CUG for valyl-tRNA)/TIS may also be used. The nucleic acids of the present invention may comprise at least one termination signal. A “termination signal" or "terminator" is comprised of the DNA sequences involved in specific termination of an RNA transcript by an RNA polymerase. Thus, a termination signal that ends the production of an RNA transcript is contemplated according to the present invention. A terminator may be necessary in vivo to achieve desirable message levels. In eukaryotic systems, a terminator region may also comprise specific DNA sequences that permit site-specific cleavage of the new transcript so as to expose a polyadenylation site. This signals a specialized endogenous polymerase to add a stretch of about 200 A residues (polyA) to the 3’ end of the transcript. RNA molecules modified with this polyA tail appear to more stable and are translated more efficiently. Thus, when the nucleic acid is for expression in eukaryotes, a terminator typically comprises a signal for the cleavage of the RNA, and it is preferred that the terminator signal promotes polyadenylation of the message. The terminator and/or polyadenylation site elements can serve to enhance message levels and to minimize read through from the cassette into other sequences. Terminators contemplated for use in the invention include any known terminator of transcription described herein or known to one of ordinary skill in the art, including but not limited to, for example, the termination sequences of genes, such as for example the bovine growth hormone terminator or viral termination sequences, such as for example the SV40 terminator. In certain embodiments, the termination signal may be a lack of transcribable or translatable sequence, such as due to a sequence truncation. The nucleic acids of the invention are capable of expressing the therapeutic protein in a given host cell. Any appropriate host cell may be used, such as mammalian, bacterial, insect, yeast, and/or plant host cells. In addition, cell-free expression systems may be used. Such expression systems and host cells are standard in the art. Typically the nucleic acid cassettes and vectors of the invention are capable of expressing the therapeutic protein in airway cells, as described herein in relation to retroviral/lentiviral (e.g. SIV) vectors of the invention. The nucleic acids of the invention may be made using any suitable process known in the art. Thus, the nucleic acids may be made using chemical synthesis techniques. Alternatively, the nucleic acids of the invention may be made using molecular biology techniques. A nucleic acid of the invention may be used in the production of a retroviral/lentiviral (e.g. SIV) vector, as described herein. By way of non-limiting example, a nucleic acid of the invention may be a plasmid which is used in the manufacture of a retroviral/lentiviral (e.g. SIV) vector. A nucleic acid of the invention may be comprised in a retroviral/lentiviral (e.g. SIV) vector. A nucleic acid of the invention may be in the form of a DNA vector, such as a DNA plasmid. The vector(s) may be an RNA vector, such as a mRNA vector or a self-amplifying RNA vector. The DNA and/or RNA vector(s) of the invention may be capable of expression in eukaryotic and/or prokaryotic cells. Typically, the DNA and/or RNA vector(s) are capable of expression in a cell of a subject, for example, a cell of a mammalian or avian subject to be immunised. Typically the nucleic acids of the invention are capable of expressing the therapeutic protein in airway cells (as described herein). A non-viral vector of the present invention may be a phage vector, such as an AAV/phage hybrid vector as described in Hajitou et al., Cell 2006; 125(2) pp. 385-398; herein incorporated by reference. Nucleic acids of the present invention (e.g. DNA or RNA vectors) may be designed in silico, and then synthesised by conventional polynucleotide synthesis techniques. Non-viral plasmids cannot replicate in the subject to be treated, as they lack the viral genetic material which hijacks the body's normal production machinery. However they are capable of replicating in appropriate host cells, such as yeasts or bacteria including E. coli, and particularly airway cells as defined herein. The term "plasmid" as used herein refers to a construction comprised of genetic material designed to direct transformation of a targeted cell. The plasmid contains a plasmid backbone. A "plasmid backbone" as used herein contains multiple genetic elements positionally and sequentially oriented with other necessary genetic elements such that the nucleic acid in the nucleic acid can be transcribed and when necessary translated in the transfected cells. The plasmid backbone can contain one or more unique restriction sites within the backbone. The plasmid may be capable of autonomous replication in a defined host or organism such that the cloned sequence is reproduced. The plasmid can confer some well-defined phenotype on the host organism which is either selectable or readily detected. The plasmid or plasmid backbone may have a linear or circular configuration. The components of a plasmid can contain, but is not limited to, a DNA molecule incorporating: (1) the plasmid backbone; (2) a sequence encoding a signal peptide; (3) a sequence encoding a therapeutic protein; and (4) regulatory elements for transcription, translation, RNA stability and replication The purpose of the plasmid in human gene therapy for the efficient delivery of nucleic acid sequences to, and expression of therapeutic proteins in, a cell or tissue. In particular, the purpose of the plasmid is to achieve high copy number, avoid potential causes of plasmid instability and provide a means for plasmid selection. As for expression, a nucleic acid of the invention contains the necessary elements for expression of the transgene comprised in the nucleic acid. Expression includes the efficient transcription of an inserted gene, nucleic acid sequence, or nucleic acid within the plasmid. A DNA plasmid may be CpG-free, or be optimised to reduce CpG dinucleotides as described herein. A DNA plasmid of the invention may be codon-optimised as described herein. Methods of preparing plasmid DNA are well known in the art. Typically, they are capable of autonomous replication in an appropriate host or producer cell. Host cells containing (e.g. transformed, transfected, or electroporated with) the plasmid may be prokaryotic or eukaryotic in nature, either stably or transiently transformed, transfected, or electroporated with the plasmid. Suitable host cells include bacterial, yeast, fungal, invertebrate, and mammalian cells. Preferably the host cell is bacterial; more preferably E. coli. Host cells can then be used in methods for the large scale production of the plasmid. The cells are grown in a suitable culture medium under favourable conditions, and the desired plasmid isolated from the cells, or from the medium in which the cells are grown, by any purification technique well known to those skilled in the art; e.g. see Sambrook et al, supra. Any appropriate delivery means can be used to deliver a non-viral vector (e.g. plasmid) of the invention to a target cell or patient. Suitable delivery means are known in the art and within the routine skill of one of ordinary skill in the art. Non-limiting examples include the use of cationic lipids, polymers (e.g. polyethyleneimine and poly-L-lysine) and electroporation. Preferably cationic lipids may be used to deliver non-viral (e.g. plasmid) vectors of the invention to target cells or to a patient. Non-limiting examples of cationic lipids suitable for use according to the invention are GL67A and lipofectamine. The cationic lipid mixture GL67A is a mixture of three components - GL67 (Cholest-5-en-3-ol (3β)-,3-[(3-aminopropyl)[4-[(3- aminopropyl)amino]butyl]carbamate], (CAS Number: 179075-30-0)), DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and DMPE-PEG5000 (1,2-Dimyristoyl-sn- Glycero-3-Phosphoethanolamine-N-[methoxy (Polyethylene glycol)5000]). These components are formulated at a 1:2:0.05 molar ratio to form GL67A. The composition of GL67A and methods for its production are disclosed in WO2013/061091, as are methods for preparing mixtures of GL67A with exemplary non-viral vectors. The contents of WO2013/061091 are herein incorporated by reference in their entirety. Lipofectamine consists of a 3:1 mixture of DOSPA (2,3-dioleoyloxy-N- [2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propaniminium trifluoroacetate) and DOPE. The invention also provides host cells comprising a nucleic acid (e.g. plasmid) of the invention. Typically a host cell is a mammalian cell, particularly a human cell or cell line. Non-limiting examples of host cells include HEK293 cells (such as HEK293F or HEK293T cells) and 293T/17 cells. Commercial cell lines suitable for the production of virus are also readily available (as described herein). Methods of Production Methods for the production of retroviral/lentiviral (e.g. SIV) vectors of the invention as also described herein. The present inventors have previously demonstrated that the use of codon-optimised gal-pol genes from SIV does not negatively impact the manufactured titre of a SIV vector pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, and can even result in an increased titre of the vector. This is described in PCT/GB2022/050524, which is herein incorporated by reference in its entirety. Further, the inventors have shown that retroviral vectors comprising a retroviral/lentiviral RNA sequence comprising (i) codon substitutions and (ii) a reduced number of modified retroviral/lentiviral open reading frames (ORFs) do not negatively impact the manufactured vector titre, transgene expression and/or integration of the retroviral/lentiviral RNA sequence into the host/target cell genome, and can even result in an increase in vector titre, transgene expression and/or integration of the retroviral/lentiviral RNA sequence. This is described in UK Application No.2212472.1, which is herein incorporated by reference in its entirety. The present inventors have now shown that retroviral/lentiviral (e.g. SIV) vectors can be produced with the endogenous RRE of the retroviral/lentiviral (e.g. SIV) genome deleted, and an intron with a retroviral/lentiviral (e.g. SIV) inserted therein introduced into the retroviral/lentiviral (e.g. SIV) genome, and that this can increase transgene expression. Accordingly, the present invention provides a method of producing a retroviral/lentiviral (e.g. SIV) vector from which (i) the endogenous RRE of the retroviral/lentiviral (e.g. SIV) genome has been deleted, and (ii) an intron with a retroviral/lentiviral (e.g. SIV) inserted therein has been introduced. A retroviral/lentiviral (e.g. SIV) may typically be pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus or with VSV-G, and which comprises a promoter and a transgene. Preferably said retroviral/lentiviral (e.g. SIV) vector is a lentiviral vector, with Simian immunodeficiency virus (SIV) vectors being particularly preferred. The method of the invention may be a scalable GMP-compatible method. The method of the invention allows the generation of retroviral/lentiviral (e.g. SIV) vectors as described herein, which exhibit high levels of transgene expression. Typically a method of the invention produces retroviral/lentiviral (e.g. SIV) vectors as described herein that exhibit increased transgene expression compared with a corresponding retroviral/lentiviral (e.g. SIV) vector which lacks an RRE-comprising intron according to the invention. Preferably, the increase in transgene expression by a retroviral/lentiviral (e.g. SIV) vector of the invention may be an increase of at least about 5-fold, an increase of at least about 10-fold, an increase of at least about 50-fold, an increase of at least about 100-fold, an increase of at least about 200-fold, an increase of at least about 500-fold, an increase of at least about 600-fold or more, typically compared with expression of the transgene by a corresponding retroviral/lentiviral (e.g. SIV) vector which lacks an RRE-comprising intron according to the invention produced by the same method. More preferably, the increase in transgene expression by a retroviral/lentiviral (e.g. SIV) vector of the invention may be at least about 10-fold, more preferably at least about 100-fold, even more preferably at least about 500-fold, typically compared with expression of the transgene by a corresponding retroviral/lentiviral (e.g. SIV) vector which lacks an RRE-comprising intron according to the invention produced by the same method. By way of non-limiting example, when the RRE-comprising intron is a β-globulin/IgG chimeric intron comprising an RRE, such as the β-globulin/IgG chimeric RRE-comprising intron of SEQ ID NO: 5, said intron may increase transgene expression by a retroviral/lentiviral (e.g. SIV) vector of the invention comprising said β-globulin/IgG chimeric RRE-comprising intron by at least 600-fold, such as by about 686-fold compared with the expression of the transgene by a corresponding retroviral/lentiviral (e.g. SIV) vector without said β-globulin/IgG chimeric RRE-comprising intron. A method of the invention typically allows the generation of retroviral/lentiviral (e.g. SIV) vectors comprising a modified retroviral/lentiviral (e.g. SIV) RNA sequence with high levels of vector integration into the host/target cell genome. Alternatively or additionally, a method of the invention may allow the generation of high titre purified retroviral/lentiviral (e.g. SIV) vectors comprising a modified retroviral/lentiviral (e.g. SIV) RNA sequence. These advantageous properties of the vectors and methods of the invention are as described in UK Application No. 2212472.1, which is herein incorporated by reference in its entirety. The production of retroviral/lentiviral (e.g. SIV) vectors typically employs one or more plasmids which provide the elements needed for the production of the vector: the genome for the retroviral/lentiviral vector, the Gag-Pol, Rev, F and HN. Multiple elements can be provided on a single plasmid. Preferably each element is provided on a separate plasmid, such that there five plasmids, one for each of the vector genome, the Gag-Pol, Rev, F and HN, respectively. Alternatively, a single plasmid may provide the Gag-Pol and Rev elements, and may be referred to as a packaging plasmid (pDNA2). The remaining elements (genome, F and HN) may be provided by separate plasmids (pDNA1, pDNA3a, pDNA3b respectively), such that four plasmids are used for the production of a retroviral/lentiviral (e.g. SIV) vector according to the invention. In the four plasmid methods, pDNA1, pDNA3a and pDNA3b may be as described herein in the context of the five-plasmid method. For retroviral/lentiviral (e.g. SIV) vectors pseudotyped with another envelope protein, such as VSV-G, rather than F and HN proteins, again a method of the invention typically employs one or more plasmids which provide the elements needed for the production of the vector: the genome for the retroviral/lentiviral vector, the Gag-Pol (pDNA2a), Rev (pDNA2b), and envelope (e.g. VSV-G) (pDNA3). Multiple elements can be provided on a single plasmid. Preferably each element is provided on a separate plasmid, such that there four plasmids, one for each of the vector genome, the Gag-Pol, Rev and envelope (e.g. VSV-G), respectively. In the four plasmid methods for VSV-G pseudotyped retroviral/lentiviral vectors, pDNA1, pDNA2a and pDNA2b may be as described herein in the context of the five-plasmid method for retroviral/lentiviral vectors pseudotyped with F and HN proteins. Alternatively, a single plasmid may provide the Gag-Pol and Rev elements, and may be referred to as a packaging plasmid (pDNA2). The remaining elements (genome and VSV-G) may be provided by separate plasmids (pDNA1 and pDNA3 respectively), such that three plasmids are used for the production of a retroviral/lentiviral (e.g. SIV) vector according to the invention. In the three plasmid methods, pDNA1 may be as described herein in the context of the five/four-plasmid methods. Preferably, the vector genome plasmid encodes all the genetic material that is packaged into the final retroviral/lentiviral vector, including the transgene. The vector genome plasmid may be designated herein as “pDNA1”, and typically comprises the transgene and the transgene promoter. As described herein, the RRE-comprising intron is typically comprised within the vector genome plasmid. The RRE-comprising intron is typically incorporated from the vector genome plasmid into the retroviral/lentiviral (e.g. SIV) RNA sequence. The other four plasmids are manufacturing plasmids encoding the Gag-Pol, Rev, F and HN proteins. These plasmids may be designated “pDNA2a”, “pDNA2b”, “pDNA3a” and “pDNA3b” respectively. Typically, the lentivirus is SIV, such as SIV1, preferably SIV-AGM. The F and HN proteins are derived from a respiratory paramyxovirus, preferably a Sendai virus. In a specific embodiment relating to AAT, the five plasmids are characterised by Figures 2A- 2F, thus pDNA1 is the pGM991 plasmid of Figure 2A, pDNA2a is the pGM691 plasmid of Figure 2B or the pGM297 plasmid of Figure 2C, pDNA2b is the pGM299 plasmid of Figure 2D, pDNA3a is the pGM301 plasmid of Figure 2E and pDNA3b is the pGM303 plasmid of Figure 2F, or variants thereof any of these plasmids (as described herein). pGM407 (as shown in Figure 2G) is an unmodified version of the vector genome plasmid from which pGM991 is derived. The plasmid as defined in Figure 2A is represented by SEQ ID NO: 30; the plasmid as defined in Figure 2B is represented by SEQ ID NO: 31; the plasmid as defined in Figure 2C is represented by SEQ ID NO: 32; the plasmid as defined in Figure 2D is represented by SEQ ID NO: 33; the plasmid as defined in Figure 2E is represented by SEQ ID NO: 34; the plasmid as defined in Figure 2F is represented by SEQ ID NO: 35; and the plasmid as defined in Figure 2G is represented by SEQ ID NO: 36. Variants (as defined herein) of these plasmids are also encompassed by the present invention. In particular, variants having at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99, 99.5 or 100%) sequence identity to any one of SEQ ID NOs: 30 to 36 are encompassed. In each of the three-, four- or five-plasmid methods of the invention all of the plasmids contribute to the formation of the final retroviral/lentiviral (e.g. SIV) vector, although only the vector genome plasmid provides nucleic acid sequence comprised in the retroviral/lentiviral (e.g. SIV) RNA sequence. During manufacture of the retroviral/lentiviral (e.g. SIV) vector, the vector genome plasmid (pDNA1) provides the enhancer/promoter, Psi, RRE-comprising intron, cPPT, mWPRE, SIN LTR, SV40 polyA (see Figure 1A), which are important for virus manufacture. Using pGM991 as a non-limiting example of a pDNA1, the CMV enhancer/promoter, SV40 polyA, colE1 Ori and KanR are involved in manufacture of the retroviral/lentiviral (e.g. SIV) vector of the invention, but are not found in the final retroviral/lentiviral (e.g. SIV) vector. The cPPT (central polypurine tract), RRE-comprising intron (inserted between hCEF and the AAT transgene), hCEF, AAT (transgene) and mWPRE from pGM991 are found in the final retroviral/lentiviral (e.g. SIV) vector. SIN LTR (long terminal repeats, SIN/IN self- inactivating) and Psi (packaging signal) may be found in the final retroviral/lentiviral (e.g. SIV) vector. In contrast, compared with pGM991, pGM407 (from which pGM991 is derived) lacks the RRE- comprising intron, but comprises the endogenous SIV RRE, which is positioned 5’ of the hCEF promoter and between the partial Gag and cPPT sequences. For other retroviral/lentiviral (e.g. SIV) vectors of the invention, corresponding elements from the other vector genome plasmids (pDNA1) are required for manufacture (but not found in the final vector), or are present in the final retroviral/lentiviral (e.g. SIV) vector. In a specific embodiment relating to pseudotyping with VSV-G, the four plasmids are characterised by Figures 2A-2F, thus pDNA1 is the pGM991 plasmid of Figure 2A, pDNA2a is the pGM691 plasmid of Figure 2B or the pGM297 plasmid of Figure 2C, pDNA2b is the pGM299 plasmid of Figure 2D, pDNA3 is the pMD2.G plasmid of Figure 2H, or variants thereof any of these plasmids (as described herein). The plasmid as defined in Figure 2A is represented by SEQ ID NO: 30; the plasmid as defined in Figure 2B is represented by SEQ ID NO: 31; the plasmid as defined in Figure 2C is represented by SEQ ID NO: 32; the plasmid as defined in Figure 2D is represented by SEQ ID NO: 33; the plasmid as defined in Figure 2H is represented by SEQ ID NO: 49. Variants (as defined herein) of these plasmids are also encompassed by the present invention. In particular, variants having at least 90% (such as at least 90, 92, 94, 95, 96, 97, 98, 99, 99.5 or 100%) sequence identity to any one of SEQ ID NOs: 30 to 33 and 49 are encompassed. The F and HN proteins from pDNA3a and pDNA3b (preferably Sendai F and HN proteins) or the VSV-G from pDNA3 are important for infection of target cells with the final retroviral/lentiviral (e.g. SIV) vector, i.e. for entry of a patient’s epithelial cells (typically lung or nasal cells as described herein). The products of the pDNA2a and pDNA2b plasmids (or pDNA2 if the Gag-Pol and Rev elements are combined in a single plasmid) are important for virus transduction, i.e. for inserting the retroviral/lentiviral (e.g. SIV) DNA into the host’s genome. The promoter, regulatory elements (such as WPRE) and transgene are important for transgene expression within the target cell(s). A method of the invention may comprise or consist of the following steps: (a) growing cells in suspension; (b) transfecting the cells with one or more plasmids; (c) adding a nuclease; (d) harvesting the retrovirus/lentivirus (e.g. SIV); (e) adding trypsin; and (f) purification of the retrovirus/lentivirus (e.g. SIV). This method may use the three-, four- or five-plasmid system described herein. Thus, for a five-plasmid method, the one or more plasmids may comprise or consist of: a vector genome plasmid pDNA1; a Gag-Pol plasmid (e.g. codon-optimised Gag-Pol plasmid), pDNA2a; a Rev plasmid, pDNA2b; a fusion (F) protein plasmid, pDNA3a; and a hemagglutinin-neuraminidase (HN) plasmid, pDNA3b. The pDNA1 may be pGM991. The pDNA2a may be pGM297 or pGM691, preferably pGM691. The pDNA2b may be pGM299. The pDNA3a may be pGM301. The pDNA3b may be pGM303. Any combination of pDNA1, pDNA2a, pDNA2b, pDNA3a and pDNA3b may be used. Preferably, the pDNA1 is pGM991; the pDNA2a is pGM691; the pDNA2b is pGM299; the pDNA3a is pGM301; and the pDNA3b is pGM303. For a four-plasmid method, the one or more plasmids may comprise or consist of: a vector genome plasmid pDNA1; a Gag-Pol plasmid (e.g. codon-optimised Gag-Pol plasmid), pDNA2a; a Rev plasmid, pDNA2b; and a VSV-G plasmid, pDNA3. The pDNA1 may be pGM991. The pDNA2a may be pGM297 or pGM691, preferably pGM691. The pDNA2b may be pGM299. The pDNA3 may be pMD2.G. Any combination of pDNA1, pDNA2a, pDNA2b, and pDNA3 may be used. Preferably, the pDNA1 is pGM991; the pDNA2a is pGM691; the pDNA2b is pGM299; the pDNA3a is pGM301; and the pDNA3 is pMD2.G. Any appropriate ratio of vector genome plasmid: Gag-Pol plasmid: Rev plasmid: F plasmid: HN plasmid may be used to further optimise (increase) the retroviral/lentiviral (e.g. SIV) titre produced. By way of non-limiting example, the ratio of vector genome plasmid: Gag-Pol plasmid: Rev plasmid: F plasmid: HN plasmid may by in the range of 10-40:-4-20:3-12:3-12:3-12, typically 15-20:7-11:4-8:4- 8:4-8, such as about 18-22:7-11:4-8:4-8:4-8, 19-21:8-10:5-7:5-7:5-7. Preferably the ratio of vector genome plasmid: Gag-Pol plasmid: Rev plasmid: F plasmid: HN plasmid is about 20:9:6:6:6. Preferably the ratio of vector genome plasmid: Gag-Pol plasmid: Rev plasmid: VSV-G plasmid is about 20:9:6:12. Steps (a)-(f) of the method are typically carried out sequentially, starting at step (a) and continuing through to step (f). The method may include one or more additional step, such as additional purification steps, buffer exchange, concentration of the retroviral/lentiviral (e.g. SIV) vector after purification, and/or formulation of the retroviral/lentiviral (e.g. SIV) vector after purification (or concentration). Each of the steps may comprise one or more sub-steps. For example, harvesting may involve one or more steps or sub-steps, and/or purification may involve one or more steps or sub-steps. Any appropriate cell type may be transfected with the one or more plasmids (e.g. the five-, four- or three- plasmids described herein) to produce a retroviral/lentiviral (e.g. SIV) vector of the invention. Typically mammalian cells, particularly human cell lines are used. Non-limiting examples of cells suitable for use in the methods of the invention are HEK293 cells (such as HEK293F or HEK293T cells) and 293T/17 cells. Commercial cell lines suitable for the production of virus are also readily available (e.g. Gibco Viral Production Cells – Catalogue Number A35347 from ThermoFisher Scientific). The cells may be grown in animal-component free media, including serum-free media. The cells may be grown in a media which contains human components. The cells may be grown in a defined media comprising or consisting of synthetically produced components. Any appropriate transfection means may be used according to the invention. Selection of appropriate transfection means is within the routine practice of one of ordinary skill in the art. By way of non-limiting example, transfection may be carried out by the use of PEIPro^TM, Lipofectamine2000^TM or Lipofectamine3000^TM. Any appropriate nuclease may be used according to the invention. Selection of appropriate nuclease is within the routine practice of one of ordinary skill in the art. Typically the nuclease is an endonuclease. By way of non-limiting example, the nuclease may be Benzonase® or Denarase®. The addition of the nuclease may be at the pre-harvest stage or at the post-harvest stage, or between harvesting steps. The gag-pol genes used in the production of a retroviral/lentiviral (e.g. SIV) vectors of the invention may be codon-optimised. Thus, the gag-pol genes within the pDNA2a plasmid may be codon-optimised. By way of non-limiting example, codon-optimised gag-pol genes may comprise or consist of the nucleic acid sequence of SEQ ID NO: 37, or a variant thereof (as defined herein). In particular, the codon-optimised gag-pol genes of the invention may comprise or consist of a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more sequence identity to SEQ ID NO: 37, preferably at least 95%, identity to SEQ ID NO: 37. The codon-optimised gag-pol genes may consist of the nucleic acid sequence of SEQ ID NO: 37. The preferred pDNA2a, pGM691, comprises the codon-optimised gag-pol genes of SEQ ID NO: 37. The gag-pol genes (e.g. SIV gag-pol genes), including codon-optimised gag-pol genes are typically operably linked to a promoter to facilitate expression of the gag-pol proteins. Any suitable promoter may be used, including those described herein in the context of promoters for the transgene. Preferably, the promoter is a CAG promoter, as used on the exemplified pGM691 plasmid. An exemplary CAG promoter is set out in SEQ ID NO: 38. The codon-optimised gag-pol genes of SEQ ID NO: 37 comprise a translational slip, and so do not form a single conventional open reading frame. Codon-optimised gag-pol genes (or nucleic acids comprising or consisting thereof) and plasmids comprising said genes or nucleic acids are advantageous in the production of retroviral/lentiviral (e.g. SIV) vectors using methods of the invention, as they allow for the production of high titre retroviral/lentiviral (e.g. SIV) vectors. Typically said codon-optimised gag-pol genes (or nucleic acids comprising or consisting thereof) and plasmids comprising said genes or nucleic acids can be used to produces a titre of retroviral/lentiviral (e.g. SIV) vector that is at least equivalent to the titre of retroviral/lentiviral (e.g. SIV) vector produced by a corresponding method which does not use codon-optimised gag-pol genes, as described herein. Codon-optimised gag-pol genes are further disclosed in PCT/GB2022/050524, which is herein incorporated by reference in its entirety. The invention also provides a retroviral/lentiviral (e.g. SIV) vector obtainable by a method of the invention. Typically, the retroviral/lentiviral (e.g. SIV) vector obtainable by a method of the invention is produced at a high-titre, as described herein. Titre may be measured in terms of transducing units, as defined here. Accordingly, the retroviral/lentiviral (e.g. SIV) vectors of the invention, including those obtainable by a method of the invention may optionally be at a titre of at least about 2.5x10⁶ TU/mL, at least about 3.0x10⁶ TU/mL, at least about 3.1x10⁶ TU/mL, at least about 3.2x10⁶ TU/mL, at least about 3.3x10⁶ TU/mL¸ at least about 3.4x10⁶ TU/mL, at least about 3.5x10⁶ TU/mL, at least about 3.6x10⁶ TU/mL, at least about 3.7x10⁶ TU/mL, at least about 3.8x10⁶ TU/mL, at least about 3.9x10⁶ TU/mL, at least about 4.0x10⁶ TU/mL or more. Preferably the retroviral/lentiviral (e.g. SIV) vector is produced at a titre of at least about 3.0x10⁶ TU/mL, or at least about 3.5x10⁶ TU/mL. The production of high-titre retroviral/lentiviral (e.g. SIV) vectors may impart other desirable properties on the resulting vector products. For example, without being bound by theory, it is believed that production at high titres without the need for intense concentration by methods such as TFF results in a higher quality vector product than retroviral/lentiviral (e.g. SIV) vectors produced by corresponding methods without the use of codon-optimised gag-pol genes (and optionally a modified vector genome plasmid), because the vectors are exposed to less shear forces which can damage the viral particles and their RNA cargo. Typically the gag-pol genes (e.g. codon-optimised gag-pol genes) used are matched to the retroviral/lentiviral vector being produced. By way of non-limiting example, when the lentiviral vector is an HIV vector, the codon-optimised gag-pol genes used are HIV gag-pol genes. By way of non- limiting example, when the lentiviral vector is an SIV vector, the codon-optimised gag-pol genes used are SIV gag-pol genes. Preferably the codon-optimised gag-pol genes used are SIV gag-pol genes. As described herein, the retroviral/lentiviral (e.g. SIV) vectors of the invention (i) lack the endogenous RRE; and (ii) comprise an RRE-comprising intron. Accordingly, the vector genome plasmid used in the production of a retroviral/lentiviral (e.g. SIV) vector of the invention may be modified to (i) delete the endogenous RRE and (ii) introduce an RRE-comprising intron. Any disclosure herein in relation to retroviral/lentiviral (e.g. SIV) vectors (i) lacking the endogenous RRE and (ii) comprising an RRE-comprising intron, applies equally and without reservation to the vector genome plasmids (pDNA1) described herein, which may be used in the production of retroviral/lentiviral (e.g. SIV) vectors of the invention. As used herein, the term “trypsin” refers to both trypsin and equivalents thereof. An equivalent enzyme is one with the same or essentially the same cleavage specificity as trypsin. Trypsin cleavage activity may be defined as cleavage C-terminal to arginine or lysine residues, typically exclusively C-terminal to arginine or lysine residues. The trypsin activity may preferably be provided by an animal origin free, recombinant enzyme such as TrypLE Select™. The addition of trypsin may be at the pre-harvest stage or at the post-harvest stage, or between harvesting steps. Any appropriate purification means may be used to purify the retroviral/lentiviral (e.g. SIV) vector. Non-limiting examples of suitable purification steps include depth/end filtration, tangential flow filtration (TFF) and chromatography. The purification step typically comprises at least on chromatography step. Non-limiting examples of chromatography steps that may be used in accordance with the invention include mixed-mode size exclusion chromatography (SEC) and/or anion exchange chromatography. Elution may be carried out with or without the use of a salt gradient, preferably without. This method may be used to produce the retroviral/lentiviral (e.g. SIV) vectors of the invention, such as those comprising a CFTR, AAT and/or FVIII gene as described herein. Alternatively, the retroviral/lentiviral (e.g. SIV) vector of the invention comprises any of the above-mentioned genes, or the genes encoding the above-mentioned proteins. The method, may use any combination of one or more of the specific plasmid constructs provided by Figures 2A-2F or 2H to provide a retroviral/lentiviral (e.g. SIV) vector of the invention. Particularly the plasmid constructs of Figures 2A, 2B and 2D-2F or 2A, 2B and 2D or 2H are used. Therapeutic Indications The retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the present invention enable high and sustained transgene expression. The retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids), and particularly the F/HN-pseudotyped retroviral/lentiviral (e.g. SIV) vectors of the invention are capable of: (i) airway transduction without disruption of epithelial integrity; (ii) persistent gene expression; (iii) lack of chronic toxicity; and (iv) efficient repeat administration. Long term/persistent stable gene expression, preferably at a therapeutically-effective level, may be achieved using repeat doses of a vector of the present invention. Alternatively, a single dose may be used to achieve the desired long-term expression. Thus, advantageously, the retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the present invention can be used in gene therapy. By way of example, the efficient airway cell uptake properties of the retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the invention make them highly suitable for treating respiratory tract diseases. The retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the invention can also be used in methods of gene therapy to promote secretion of therapeutic proteins. By way of further example, the invention provides secretion of therapeutic proteins into the lumen of the respiratory tract or the circulatory system. Thus, administration of a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) of the invention and its uptake by airway cells may enable the use of the lungs (or nose or airways) as a “factory” to produce a therapeutic protein that is then secreted and enters the general circulation at therapeutic levels, where it can travel to cells/tissues of interest to elicit a therapeutic effect. In contrast to intracellular or membrane proteins, the production of such secreted proteins does not rely on specific disease target cells being transduced, which is a significant advantage and achieves high levels of protein expression. Thus, other diseases which are not respiratory tract diseases, such as cardiovascular diseases and blood disorders, particularly blood clotting deficiencies, can also be treated by the retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the present invention. Retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the invention can effectively treat a disease by providing a transgene for the correction of the disease. For example, inserting a functional copy of the CFTR gene to ameliorate or prevent lung disease in CF patients, independent of the underlying mutation. Accordingly, retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the invention may be used to treat cystic fibrosis (CF), typically by gene therapy with a CFTR transgene as described herein. As another example, retroviral/lentiviral (e.g. SIV) vectors and nucleic acids (e.g. plasmids) of the invention may be used to treat Alpha-1 Antitrypsin (AAT) deficiency, typically by gene therapy with a AAT transgene as described herein. AAT is a secreted anti-protease that is produced mainly in the liver and then trafficked to the lung, with smaller amounts also being produced in the lung itself. The main function of AAT is to bind and neutralise/inhibit neutrophil elastase. Gene therapy with AAT according to the present invention is relevant to AAT deficient patient, as well as in other lung diseases such as CF or chronic obstructive pulmonary disease (COPD), and offers the opportunity to overcome some of the problems encountered by conventional enzyme replacement therapy (in which AAT isolated from human blood and administered intravenously every week), providing stable, long-lasting expression in the target tissue (lung/nasal epithelium), ease of administration and unlimited availability. Transduction with a retroviral/lentiviral (e.g. SIV) vector of the invention or transfection with a nucleic acid (e.g. plasmid) of the invention may lead to secretion of the recombinant protein into the lumen of the lung as well as into the circulation. One benefit of this is that the therapeutic protein reaches the interstitium. AAT gene therapy may therefore also be beneficial in other disease indications, non-limiting examples of which include type 1 and type 2 diabetes, acute myocardial infarction, ischemic heart disease, rheumatoid arthritis, inflammatory bowel disease, transplant rejection, graft versus host (GvH) disease, multiple sclerosis, liver disease, cirrhosis, vasculitides and infections, such as bacterial and/or viral infections. AAT has numerous other anti-inflammatory and tissue-protective effects, for example in pre- clinical models of diabetes, graft versus host disease and inflammatory bowel disease. The production of AAT in the lung and/or nose following transduction according to the present invention may, therefore, be more widely applicable, including to these indications. Other examples of diseases that may be treated with gene therapy of a secreted protein according to the present invention include cardiovascular diseases and blood disorders, particularly blood clotting deficiencies such as haemophilia (A, B or C), von Willebrand disease and Factor VII deficiency. Other examples of diseases or disorders to be treated include Primary Ciliary Dyskinesia (PCD), acute lung injury, Surfactant Protein B (SFTB) deficiency, Pulmonary Alveolar Proteinosis (PAP), Chronic Obstructive Pulmonary Disease (COPD) and/or inflammatory, infectious, immune or metabolic conditions, such as lysosomal storage diseases. Accordingly, the invention provides a method of treating a disease, the method comprising administering a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) of the invention to a subject. Typically the retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) is produced using a method of the present invention. Any disease described herein may be treated according to the invention. In particular, the invention provides a method of treating a lung disease using a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) of the invention. The disease to be treated may be a chronic disease. Preferably, a method of treating CF is provided. The invention also provides a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) as described herein for use in a method of treating a disease. Typically the retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) is produced using a method of the present disclosure. Any disease described herein may be treated according to the invention. In particular, the invention provides a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) of the invention for use in a method of treating a lung disease. The disease to be treated may be a chronic disease. Preferably, a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) for use in treating CF is provided. The invention also provides the use of a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) as described herein in the manufacture of a medicament for use in a method of treating a disease. Typically the retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) is produced using a method of the present disclosure. Any disease described herein may be treated according to the invention. In particular, the invention provides the use of a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) of the invention for the manufacture of a medicament for use in a method of treating a lung disease. The disease to be treated may be a chronic disease. Preferably, the use of a retroviral/lentiviral (e.g. SIV) vector or nucleic acid (e.g. plasmid) in the manufacture of a medicament for use in a method of treating CF is provided. Formulation and administration The retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) of the invention may be administered in any dosage appropriate for achieving the desired therapeutic effect. Appropriate dosages may be determined by a clinician or other medical practitioner using standard techniques and within the normal course of their work. Non-limiting examples of suitable dosages of a retroviral/lentiviral (e.g. SIV) vector include 1x10⁸ transduction units (TU), 1x10⁹ TU, 1x10¹⁰ TU, 1x10¹¹ TU or more. The invention also provides compositions comprising the retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) described above, and a pharmaceutically-acceptable carrier. Non- limiting examples of pharmaceutically acceptable carriers include water, saline, and phosphate- buffered saline. In some embodiments, however, the composition is in lyophilized form, in which case it may include a stabilizer, such as bovine serum albumin (BSA). In some embodiments, it may be desirable to formulate the composition with a preservative, such as thiomersal or sodium azide, to facilitate long-term storage. The retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) of the invention may be administered by any appropriate route. It may be desired to direct the compositions of the present invention (as described above) to the respiratory system of a subject. Efficient transmission of a therapeutic/prophylactic composition or medicament to the site of infection in the respiratory tract may be achieved by oral or intra-nasal administration, for example, as aerosols (e.g. nasal sprays), or by catheters. Typically the retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) of the invention are stable in clinically relevant nebulisers, inhalers (including metered dose inhalers), catheters and aerosols, etc. Typically, therefore, the retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) of the invention are formulated for administration to the lungs by any appropriate means, e.g. they may be formulated for intratracheal administration, intranasal administration, aerosol delivery, or direct injection or delivery to the lungs (e.g. delivered by catheter). Other modes of delivery, e.g. intravenous delivery, are also encompassed by the invention. In some embodiments the nose is a preferred production site for a therapeutic protein using a retroviral/lentiviral (e.g. SIV) vector and/or nucleic acid (e.g. plasmid) of the invention for at least one of the following reasons: (i) extracellular barriers such as inflammatory cells and sputum are less pronounced in the nose; (ii) ease of vector administration; (iii) smaller quantities of vector/nucleic acid required; and (iv) ethical considerations. Thus, transduction of nasal epithelial cells with a retroviral/lentiviral (e.g. SIV) vector or transfection with a nucleic acid (e.g. plasmid) of the invention may result in efficient (high-level) and long-lasting expression of the therapeutic transgene of interest. Accordingly, nasal administration of a retroviral/lentiviral (e.g. SIV) vector or a nucleic acid (e.g. plasmid) of the invention may be preferred. Formulations for intra-nasal administration may be in the form of nasal droplets or a nasal spray. An intra-nasal formulation may comprise droplets having approximate diameters in the range of 100-5000 µm, such as 500-4000 µm, 1000-3000 µm or 100-1000 µm. Alternatively, in terms of volume, the droplets may be in the range of about 0.001-100 µl, such as 0.1-50 µl or 1.0-25 µl, or such as 0.001-1 µl. The aerosol formulation may take the form of a powder, suspension or solution. The size of aerosol particles is relevant to the delivery capability of an aerosol. Smaller particles may travel further down the respiratory airway towards the alveoli than would larger particles. In one embodiment, the aerosol particles have a diameter distribution to facilitate delivery along the entire length of the bronchi, bronchioles, and alveoli. Alternatively, the particle size distribution may be selected to target a particular section of the respiratory airway, for example the alveoli. In the case of aerosol delivery of the medicament, the particles may have diameters in the approximate range of 0.1-50 µm, preferably 1-25 µm, more preferably 1-5 µm. Aerosol particles may be for delivery using a nebulizer (e.g. via the mouth) or nasal spray. An aerosol formulation may optionally contain a propellant and/or surfactant. The formulation of pharmaceutical aerosols is routine to those skilled in the art, see for example, Sciarra, J. in Remington's Pharmaceutical Sciences (supra). The agents may be formulated as solution aerosols, dispersion or suspension aerosols of dry powders, emulsions or semisolid preparations. The aerosol may be delivered using any propellant system known to those skilled in the art. The aerosols may be applied to the upper respiratory tract, for example by nasal inhalation, or to the lower respiratory tract or to both. The part of the lung that the medicament is delivered to may be determined by the disorder. Compositions comprising a vector of the invention, in particular where intranasal delivery is to be used, may comprise a humectant. This may help reduce or prevent drying of the mucus membrane and to prevent irritation of the membranes. Suitable humectants include, for instance, sorbitol, mineral oil, vegetable oil and glycerol; soothing agents; membrane conditioners; sweeteners; and combinations thereof. The compositions may comprise a surfactant. Suitable surfactants include non-ionic, anionic and cationic surfactants. Examples of surfactants that may be used include, for example, polyoxyethylene derivatives of fatty acid partial esters of sorbitol anhydrides, such as for example, Tween 80, Polyoxyl 40 Stearate, Polyoxy ethylene 50 Stearate, fusieates, bile salts and Octoxynol. In some cases after an initial administration a subsequent administration of a retroviral/lentiviral (e.g. SIV) vector and/or a nucleic acid (e.g. plasmid) may be performed. The administration may, for instance, be at least a week, two weeks, a month, two months, six months, a year or more after the initial administration. In some instances, a retroviral/lentiviral (e.g. SIV) vector and/or a nucleic acid (e.g. plasmid) of the invention may be administered at least once a week, once a fortnight, once a month, every two months, every six months, annually or at longer intervals. Preferably, administration is every six months, more preferably annually. The retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) may, for instance, be administered at intervals dictated by when the effects of the previous administration are decreasing. Any two or more retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) of the invention may be administered separately, sequentially or simultaneously. Thus two or more retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids), wherein at least one retroviral/lentiviral (e.g. SIV) vector and/or nucleic acid (e.g. plasmid) is a retroviral/lentiviral (e.g. SIV) vector and/or nucleic acid (e.g. plasmid) of the invention, may be administered separately, simultaneously or sequentially. In particular two or more retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) of the invention may be administered in such a manner. The two may be administered in the same or different compositions. In a preferred instance, the two retroviral/lentiviral (e.g. SIV) vectors and/or nucleic acids (e.g. plasmids) may be delivered in the same composition. Methods to Differentiate between Retroviral/Lentiviral Vectors and Transgene mRNA Conventional methods for the production of retroviral/lentiviral (e.g. SIV) vectors produce retroviral/lentiviral (e.g. SIV) vectors, which when transcribed, produce an mRNA which is identical in sequence to the retroviral/lentiviral (e.g. SIV) genome. Thus, it is not possible to distinguish between the retroviral/lentiviral (e.g. SIV) vector genome (the prodrug) and the transcribed mRNA (which will then be translated to produce the therapeutic protein). In contrast, the present invention relates to retroviral/lentiviral (e.g. SIV) vectors which contain an RRE-comprising intron, as described herein. Thus, the retroviral/lentiviral (e.g. SIV) genome (the prodrug) contains the sequence of an RRE-comprising intron. This RRE-comprising intron is spliced out during transcription of the retroviral/lentiviral (e.g. SIV) genome, resulting in mRNA which lacks the RRE-comprising intron, and hence has a different nucleic acid sequence compared with the retroviral/lentiviral (e.g. SIV) genome from which it was derived. The different sequences of the retroviral/lentiviral (e.g. SIV) genome and the transcribed mRNA allow for the development of specific PCR- and in situ hybridisation-based assays that detect and quantify the different nucleic acid sequences, allowing for the retroviral/lentiviral (e.g. SIV) genome and the transcribed mRNA to each be quantified. Thus, the invention provides a means of discriminating between the retroviral/lentiviral (e.g. SIV) vector genome (the prodrug) and the mRNA (first step towards the active therapeutic). This may be useful, for example, during the production of the retroviral/lentiviral (e.g. SIV) vector, during its use in vitro and/or for evaluating clinical efficacy of the retroviral/lentiviral (e.g. SIV) vector. Accordingly, the present invention provides a method for differentiating between a retroviral/lentiviral (e.g. SIV) vector and a transgene expressed by said retroviral vector, said method comprising or consisting of the steps of (a) transfecting cells with a retroviral/lentiviral (e.g. SIV) vector of the invention; (b) culturing the cells to allow transgene expression by the retroviral/lentiviral (e.g. SIV) vector; and (c) quantifying RNA within the cells; wherein (i) the amount of RNA comprising the intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted corresponds to the copy number of the retroviral/lentiviral (e.g. SIV) vector; and (ii) the amount of RNA lacking the intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted corresponds to the amount of transgene mRNA. The present invention provides a method for differentiating between a retroviral/lentiviral (e.g. SIV) vector and mRNA transcribed from said retroviral/lentiviral (e.g. SIV) vector, said method comprising or consisting of the steps of (a) transfecting cells with a retroviral/lentiviral (e.g. SIV) vector of the invention; (b) culturing the cells to allow transcription of the genome of the retroviral/lentiviral (e.g. SIV) vector; and (c) quantifying RNA within the cells; wherein (i) the amount of RNA comprising the intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted corresponds to the copy number of the retroviral/lentiviral (e.g. SIV) vector; and (ii) the amount of RNA lacking the intron into which a retroviral/lentiviral (e.g. SIV) RRE has been inserted corresponds to the amount of mRNA transcribed from the retroviral/lentiviral (e.g. SIV) vector genome, and wherein optionally the amount of mRNA transcribed corresponds to the level of transgene expression. Said methods may involve the quantification of RNA by any appropriate technique, examples of which are known in the art and may be selected by a skilled person without undue burden. Preferably, said method may involve the quantification of RNA by a PCR-based and/or in situ hybridisation-based assay. Such PCR-based methods may comprise the use of two sets of primer pairs. The first primer pair includes one primer which binds to a sequence outside the intron and another primer which binds to a sequence inside the intron. This first primer pair is capable of detecting and quantifying non-spliced retroviral/lentiviral (e.g. SIV) vectors. The second primer pair comprises two primers which bind outside of the intron on either side and, therefore only quantifies spliced retroviral/lentiviral (e.g. SIV) vectors. As the retroviral/lentiviral (e.g. SIV) mRNA is spliced, the second primer pair is specific for the mRNA transcribed from the retroviral/lentiviral (e.g. SIV) vector while the first primer pair will detect retroviral/lentiviral (e.g. SIV) vector genomes and integrated retroviral/lentiviral (e.g. SIV) DNA. SEQUENCE HOMOLOGY Any of a variety of sequence alignment methods can be used to determine percent identity, including, without limitation, global methods, local methods and hybrid methods, such as, e.g., segment approach methods. Protocols to determine percent identity are routine procedures within the scope of one skilled in the art. Global methods align sequences from the beginning to the end of the molecule and determine the best alignment by adding up scores of individual residue pairs and by imposing gap penalties. Non-limiting methods include, e.g., CLUSTAL W, see, e.g., Julie D. Thompson et al., CLUSTAL W: Improving the Sensitivity of Progressive Multiple Sequence Alignment Through Sequence Weighting, Position- Specific Gap Penalties and Weight Matrix Choice, 22(22) Nucleic Acids Research 4673-4680 (1994); and iterative refinement, see, e.g., Osamu Gotoh, Significant Improvement in Accuracy of Multiple Protein. Sequence Alignments by Iterative Refinement as Assessed by Reference to Structural Alignments, 264(4) J. MoI. Biol. 823-838 (1996). Local methods align sequences by identifying one or more conserved motifs shared by all of the input sequences. Non-limiting methods include, e.g., Match-box, see, e.g., Eric Depiereux and Ernest Feytmans, Match- Box: A Fundamentally New Algorithm for the Simultaneous Alignment of Several Protein Sequences, 8(5) CABIOS 501 -509 (1992); Gibbs sampling, see, e.g., C. E. Lawrence et al., Detecting Subtle Sequence Signals: A Gibbs Sampling Strategy for Multiple Alignment, 262(5131 ) Science 208-214 (1993); Align-M, see, e.g., Ivo Van WaIIe et al., Align-M - A New Algorithm for Multiple Alignment of Highly Divergent Sequences, 20(9) Bioinformatics:1428-1435 (2004). Thus, percent sequence identity is determined by conventional methods. See, for example, Altschul et al., Bull. Math. Bio.48: 603-16, 1986 and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915-19, 1992. Briefly, two amino acid sequences are aligned to optimize the alignment scores using a gap opening penalty of 10, a gap extension penalty of 1, and the "blosum 62" scoring matrix of Henikoff and Henikoff (ibid.) as shown below (amino acids are indicated by the standard one-letter codes). The "percent sequence identity" between two or more nucleic acid or amino acid sequences is a function of the number of identical positions shared by the sequences. Thus, % identity may be calculated as the number of identical nucleotides / amino acids divided by the total number of nucleotides / amino acids, multiplied by 100. Calculations of % sequence identity may also take into account the number of gaps, and the length of each gap that needs to be introduced to optimize alignment of two or more sequences. Sequence comparisons and the determination of percent identity between two or more sequences can be carried out using specific mathematical algorithms, such as BLAST, which will be familiar to a skilled person. ALIGNMENT SCORES FOR DETERMINING SEQUENCE IDENTITY A R N D C Q E G H I L K M F P S T W Y V A 4 R -1 5 N -2 0 6 D -2 -2 1 6 C 0 -3 -3 -3 9 Q -1 1 0 0 -3 5 E -1 0 0 2 -4 2 5 G 0 -2 0 -1 -3 -2 -2 6 H -2 0 1 -1 -3 0 0 -2 8 I -1 -3 -3 -3 -1 -3 -3 -4 -3 4 L -1 -2 -3 -4 -1 -2 -3 -4 -3 2 4 K -1 2 0 -1 -3 1 1 -2 -1 -3 -2 5 M -1 -1 -2 -3 -1 0 -2 -3 -2 1 2 -1 5 F -2 -3 -3 -3 -2 -3 -3 -3 -1 0 0 -3 0 6 P -1 -2 -2 -1 -3 -1 -1 -2 -2 -3 -3 -1 -2 -4 7 S 1 -1 1 0 -1 0 0 0 -1 -2 -2 0 -1 -2 -1 4 T 0 -1 0 -1 -1 -1 -1 -2 -2 -1 -1 -1 -1 -2 -1 1 5 W -3 -3 -4 -4 -2 -2 -3 -2 -2 -3 -2 -3 -1 1 -4 -3 -211 Y -2 -2 -2 -3 -2 -1 -2 -3 2 -1 -1 -2 -1 3 -3 -2 -2 2 7 V 0 -3 -3 -3 -1 -2 -2 -3 -3 3 1 -2 1 -1 -2 -2 0 -3 -1 4 The percent identity is then calculated as: Total number of identical matches __________________________________________ x 100 [length of the longer sequence plus the number of gaps introduced into the longer sequence in order to align the two sequences] Substantially homologous polypeptides are characterized as having one or more amino acid substitutions, deletions or additions. These changes are preferably of a minor nature, that is conservative amino acid substitutions (as described herein) and other substitutions that do not significantly affect the folding or activity of the polypeptide; small deletions, typically of one to about 30 amino acids; and small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or an affinity tag. In addition to the 20 standard amino acids, non-standard amino acids (such as 4- hydroxyproline, 6-N-methyl lysine, 2-aminoisobutyric acid, isovaline and α -methyl serine) may be substituted for amino acid residues of the polypeptides of the present invention. A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code, and unnatural amino acids may be substituted for polypeptide amino acid residues. The polypeptides of the present invention can also comprise non-naturally occurring amino acid residues. Non-naturally occurring amino acids include, without limitation, trans-3-methylproline, 2,4- methano-proline, cis-4-hydroxyproline, trans-4-hydroxy-proline, N-methylglycine, allo-threonine, methyl-threonine, hydroxy-ethylcysteine, hydroxyethylhomo-cysteine, nitro-glutamine, homoglutamine, pipecolic acid, tert-leucine, norvaline, 2-azaphenylalanine, 3-azaphenyl-alanine, 4- azaphenyl-alanine, and 4-fluorophenylalanine. Several methods are known in the art for incorporating non-naturally occurring amino acid residues into proteins. For example, an in vitro system can be employed wherein nonsense mutations are suppressed using chemically aminoacylated suppressor tRNAs. Methods for synthesizing amino acids and aminoacylating tRNA are known in the art. Transcription and translation of plasmids containing nonsense mutations is carried out in a cell free system comprising an E. coli S30 extract and commercially available enzymes and other reagents. Proteins are purified by chromatography. See, for example, Robertson et al., J. Am. Chem. Soc. 113:2722, 1991; Ellman et al., Methods Enzymol. 202:301, 1991; Chung et al., Science 259:806-9, 1993; and Chung et al., Proc. Natl. Acad. Sci. USA 90:10145-9, 1993). In a second method, translation is carried out in Xenopus oocytes by microinjection of mutated mRNA and chemically aminoacylated suppressor tRNAs (Turcatti et al., J. Biol. Chem. 271:19991-8, 1996). Within a third method, E. coli cells are cultured in the absence of a natural amino acid that is to be replaced (e.g., phenylalanine) and in the presence of the desired non-naturally occurring amino acid(s) (e.g., 2-azaphenylalanine, 3- azaphenylalanine, 4-azaphenylalanine, or 4-fluorophenylalanine). The non-naturally occurring amino acid is incorporated into the polypeptide in place of its natural counterpart. See, Koide et al., Biochem. 33:7470-6, 1994. Naturally occurring amino acid residues can be converted to non-naturally occurring species by in vitro chemical modification. Chemical modification can be combined with site-directed mutagenesis to further expand the range of substitutions (Wynn and Richards, Protein Sci.2:395-403, 1993). A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code, non-naturally occurring amino acids, and unnatural amino acids may be substituted for amino acid residues of polypeptides of the present invention. Essential amino acids in the polypeptides of the present invention can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham and Wells, Science 244: 1081-5, 1989). Sites of biological interaction can also be determined by physical analysis of structure, as determined by such techniques as nuclear magnetic resonance, crystallography, electron diffraction or photoaffinity labelling, in conjunction with mutation of putative contact site amino acids. See, for example, de Vos et al., Science 255:306-12, 1992; Smith et al., J. Mol. Biol.224:899-904, 1992; Wlodaver et al., FEBS Lett.309:59-64, 1992. The identities of essential amino acids can also be inferred from analysis of homologies with related components (e.g. the translocation or protease components) of the polypeptides of the present invention. Multiple amino acid substitutions can be made and tested using known methods of mutagenesis and screening, such as those disclosed by Reidhaar-Olson and Sauer (Science 241:53-7, 1988) or Bowie and Sauer (Proc. Natl. Acad. Sci. USA 86:2152-6, 1989). Briefly, these authors disclose methods for simultaneously randomizing two or more positions in a polypeptide, selecting for functional polypeptide, and then sequencing the mutagenized polypeptides to determine the spectrum of allowable substitutions at each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem. 30:10832-7, 1991; Ladner et al., U.S. Patent No. 5,223,409; Huse, WIPO Publication WO 92/06204) and region-directed mutagenesis (Derbyshire et al., Gene 46:145, 1986; Ner et al., DNA 7:127, 1988). Multiple amino acid substitutions can be made and tested using known methods of mutagenesis and screening, such as those disclosed by Reidhaar-Olson and Sauer (Science 241:53-7, 1988) or Bowie and Sauer (Proc. Natl. Acad. Sci. USA 86:2152-6, 1989). Briefly, these authors disclose methods for simultaneously randomizing two or more positions in a polypeptide, selecting for functional polypeptide, and then sequencing the mutagenized polypeptides to determine the spectrum of allowable substitutions at each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem. 30:10832-7, 1991; Ladner et al., U.S. Patent No. 5,223,409; Huse, WIPO Publication WO 92/06204) and region-directed mutagenesis (Derbyshire et al., Gene 46:145, 1986; Ner et al., DNA 7:127, 1988). SEQUENCE INFORMATION SEQ ID NO: 1 SIV RRE sequence SEQ ID NO: 2 β-globulin/IgG chimeric intron splice donor site SEQ ID NO: 3 β-globulin/IgG chimeric intron splice acceptor site SEQ ID NO: 4 β-globulin/IgG chimeric intron SEQ ID NO: 5 β-globin/IgG chimeric intron comprising a SIV RRE SEQ ID NO: 6 Exemplified AAT transgene (SERPINA1) SEQ ID NO: 7 Complementary strand to the exemplified AAT transgene SEQ ID NO: 8 Exemplified AAT polypeptide SEQ ID NO: 9 Exemplified FVIII transgene (N6) SEQ ID NO: 10 Exemplified FVIII transgene (V3) SEQ ID NO: 11 Complementary strand to the exemplified FVIII transgene (N6) SEQ ID NO: 12 Complementary strand to the exemplified FVIII transgene (V3) SEQ ID NO: 13 Exemplified FVIII polypeptide (N6) SEQ ID NO: 14 Exemplified FVIII polypeptide (V3) SEQ ID NO: 15 Exemplified CFTR transgene (soCFTR2) SEQ ID NO: 16 Exemplified CFTR polypeptide SEQ ID NO: 17 Exemplified hGM-CSF transgene SEQ ID NO: 18 Exemplified hGM-CSF polypeptide SEQ ID NO: 19 Exemplified mGM-CSF transgene SEQ ID NO: 20 Exemplified mGM-CSF polypeptide SEQ ID NO: 21 Exemplified Human DCN (Decorin) transgene SEQ ID NO: 22 Exemplified Human Decorin polypeptide SEQ ID NO: 23 Exemplified Human TRIM72 transgene SEQ ID NO: 24 Exemplified Human TRIM72 polypeptide SEQ ID NO: 25 Exemplified Human ABCA3 (ABCA3) transgene SEQ ID NO: 26 Exemplified Human ABCA3 polypeptide SEQ ID NO: 27 Exemplified hCEF promoter SEQ ID NO: 28 Exemplified CMV promoter SEQ ID NO: 29 Exemplified EF1a promoter SEQ ID NO: 30 Plasmid as defined in Figure 2A (pDNA1 pGM991) SEQ ID NO: 31 Plasmid as defined in Figure 2B (pDNA1 pGM691) SEQ ID NO: 32 Plasmid as defined in Figure 2C (pDNA2a pGM297) SEQ ID NO: 33 Plasmid as defined in Figure 2D (pDNA2b pGM299) SEQ ID NO: 34 Plasmid as defined in Figure 2E (pDNA3a pGM301) SEQ ID NO: 35 Plasmid as defined in Figure 2F (pDNA3b pGM303) SEQ ID NO: 36 Plasmid as defined in Figure 2G (pDNA2a pGM407) SEQ ID NO: 37 Codon-optimised SIV gag-pol nucleic acid sequence (from pGM691) SEQ ID NO: 38 Exemplary CAG promoter SEQ ID NO: 39 Exemplified WPRE component (mWPRE) SEQ ID NO: 40 Exemplified Human SFTPB transgene SEQ ID NO: 41 Exemplified Human SFTPB polypeptide SEQ ID NO: 42 Exemplified Human ADAMTS13 transgene SEQ ID NO: 43 Exemplified Human ADAMTS13 polypeptide SEQ ID NO: 44 Exemplified rSIV Rev protein SEQ ID NO: 45 oIC001 DNA forward primer SEQ ID NO: 46 oIC002 DNA and RNA reverse primer SEQ ID NO: 47 oIC105 RNA forward primer SEQ ID NO: 48 CAGGS promoter comprising a chicken β-actin splice donor and a rabbit β-globulin splice acceptor SEQ ID NO: 49 Plasmid as defined in Figure 2H (pDNA3 pMD2.G) SEQ ID NO: 50 HIV RRE sequence SEQ ID NO: 1 SIV RRE sequence CCGTTTGTGCTAGGGTTCTTAGGCTTCTTGGGGGCTGCTGGAACTGCAATGGGAGCAGCGGCGACAGC CCTGACGGTCCAGTCTCAGCATTTGCTTGCTGGGATACTGCAGCAGCAGAAGAATCTGCTGGCGGCTG TGGAGGCTCAACAGCAGATGTTGAAGCTGACCATTTGGGGTGTTAAAAACCTCAATGCCCGCGTCACA GCCCTTGAGAAGTACCTAGAGGATCAGGCACGACTAAACTCCTGGGGGTGCGCATGGAAACAAGTATG TCATACCACAGTGGAGTGGCCCTGGACAAATCGGACTCCGGATTGGCAAAATATGACTTGGTTGGAGT GGGAAAGACAAATAGCTGATTTGGAAAGCAACATTACGAGACAATTAGTGAAGGCTAGAGAACAAGAG GAAAAGAATCTAGATGCCTATCAGAAGTTAACTAGTTGGTCAGATTTCTGGTCTTGGTTCGATTTCTC AAAATGGCTTAACATTTTAAAAATGGGATTTTTAGTAATAGTAGGAATAATAGGGTTAAGATTACTTT ACACAGTATATGGATGTATAGTGAGGGTTAGGCAGGGATATGTTCCTCTATCTCCACAGATCCATAT SEQ ID NO: 2 β-globulin/IgG chimeric intron splice donor site TGAGTTTAAGGTAAGT SEQ ID NO: 3 β-globulin/IgG chimeric intron splice acceptor site CTCTCCACAG SEQ ID NO: 4 β-globulin/IgG chimeric intron GTAAGTATCAAGGTTACAAGACAGGTTTAAGGAGACCAATAGAAACTGGGCTTGTCGAGACAGAGAAG ACTCTTGCGTTTCTGATAGGCACCTATTGGTCTTACTGACATCCACTTTGCCTTTCTCTCCACAG SEQ ID NO: 5 β-globin/IgG chimeric intron comprising a SIV RRE GTAAGTATCAAGGTTACAAGACAGGTTTAAGGAGACCAATAGAAACTGGGCTTGTCGAGACAGAGAAG ACTCTTGCGTTTCTGATAGGCACGCGGCCGCCCGTTTGTGCTAGGGTTCTTAGGCTTCTTGGGGGCTG CTGGAACTGCAATGGGAGCAGCGGCGACAGCCCTGACGGTCCAGTCTCAGCATTTGCTTGCTGGGATA CTGCAGCAGCAGAAGAATCTGCTGGCGGCTGTGGAGGCTCAACAGCAGATGTTGAAGCTGACCATTTG GGGTGTTAAAAACCTCAATGCCCGCGTCACAGCCCTTGAGAAGTACCTAGAGGATCAGGCACGACTAA ACTCCTGGGGGTGCGCATGGAAACAAGTATGTCATACCACAGTGGAGTGGCCCTGGACAAATCGGACT CCGGATTGGCAAAATATGACTTGGTTGGAGTGGGAAAGACAAATAGCTGATTTGGAAAGCAACATTAC GAGACAATTAGTGAAGGCTAGAGAACAAGAGGAAAAGAATCTAGATGCCTATCAGAAGTTAACTAGTT GGTCAGATTTCTGGTCTTGGTTCGATTTCTCAAAATGGCTTAACATTTTAAAAATGGGATTTTTAGTA ATAGTAGGAATAATAGGGTTAAGATTACTTTACACAGTATATGGATGTATAGTGAGGGTTAGGCAGGG ATATGTTCCTCTATCTCCACAGATCCATATGCGGCCGCCTATTGGTCTTACTGACATCCACTTTGCCT TTCTCTCCACAG Underlined = β-globin/IgG chimeric intron Double underlined = NotI restriction sites Italicised = SIV RRE sequence SEQ ID NO: 6 Exemplified AAT transgene (SERPINA1) atgcccagct ctgtgtcctg gggcattctg ctgctggctg gcctgtgctg tctggtgcct 60 gtgtccctgg ctgaggaccc tcagggggat gctgcccaga aaacagacac ctcccaccat 120 gaccaggacc accccacctt caacaagatc acccccaacc tggcagagtt tgccttcagc 180 ctgtacagac agctggccca ccagagcaac agcaccaaca tctttttcag ccctgtgtcc 240 attgccacag cctttgccat gctgagcctg ggcaccaagg ctgacaccca tgatgagatc 300 ctggaaggcc tgaacttcaa cctgacagag atccctgagg cccagatcca tgagggcttc 360 caggaactgc tgagaaccct gaaccagcca gacagccagc tgcagctgac aacaggcaat 420 gggctgttcc tgtctgaggg cctgaagctg gtggacaagt ttctggaaga tgtgaagaag 480 ctgtaccact ctgaggcctt cacagtgaac tttggggaca cagaagaggc caagaaacag 540 atcaatgact atgtggaaaa gggcacccag ggcaagattg tggaccttgt gaaagagctg 600 gacagggaca ctgtgtttgc ccttgtgaac tacatcttct tcaagggcaa gtgggagagg 660 ccctttgaag tgaaggacac tgaggaagag gacttccatg tggaccaagt gaccacagtg 720 aaggtgccaa tgatgaagag actggggatg ttcaatatcc agcactgcaa gaaactgagc 780 agctgggtgc tgctgatgaa gtacctgggc aatgctacag ccatattctt tctgcctgat 840 gagggcaagc tgcagcacct ggaaaatgag ctgacccatg acatcatcac caaatttctg 900 gaaaatgagg acagaagatc tgccagcctg catctgccca agctgagcat cacaggcaca 960 tatgacctga agtctgtgct gggacagctg ggaatcacca aggtgttcag caatggggca 1020 gacctgagtg gagtgacaga ggaagcccct ctgaagctgt ccaaggctgt gcacaaggca 1080 gtgctgacca ttgatgagaa gggcacagag gctgctgggg ccatgtttct ggaagccatc 1140 cccatgtcca tccccccaga agtgaagttc aacaagccct ttgtgttcct gatgattgag 1200 cagaacacca agagccccct gttcatgggc aaggttgtga accccaccca gaaatga 1257 SEQ ID NO: 7 Complementary strand to the exemplified AAT transgene tacgggtcga gacacaggac cccgtaagac gacgaccgac cggacacgac agaccacgga 60 cacagggacc gactcctggg agtcccccta cgacgggtct tttgtctgtg gagggtggta 120 ctggtcctgg tggggtggaa gttgttctag tgggggttgg accgtctcaa acggaagtcg 180 gacatgtctg tcgaccgggt ggtctcgttg tcgtggttgt agaaaaagtc gggacacagg 240 taacggtgtc ggaaacggta cgactcggac ccgtggttcc gactgtgggt actactctag 300 gaccttccgg acttgaagtt ggactgtctc tagggactcc gggtctaggt actcccgaag 360 gtccttgacg actcttggga cttggtcggt ctgtcggtcg acgtcgactg ttgtccgtta 420 cccgacaagg acagactccc ggacttcgac cacctgttca aagaccttct acacttcttc 480 gacatggtga gactccggaa gtgtcacttg aaacccctgt gtcttctccg gttctttgtc 540 tagttactga tacacctttt cccgtgggtc ccgttctaac acctggaaca ctttctcgac 600 ctgtccctgt gacacaaacg ggaacacttg atgtagaaga agttcccgtt caccctctcc 660 gggaaacttc acttcctgtg actccttctc ctgaaggtac acctggttca ctggtgtcac 720 ttccacggtt actacttctc tgacccctac aagttatagg tcgtgacgtt ctttgactcg 780 tcgacccacg acgactactt catggacccg ttacgatgtc ggtataagaa agacggacta 840 ctcccgttcg acgtcgtgga ccttttactc gactgggtac tgtagtagtg gtttaaagac 900 cttttactcc tgtcttctag acggtcggac gtagacgggt tcgactcgta gtgtccgtgt 960 atactggact tcagacacga ccctgtcgac ccttagtggt tccacaagtc gttaccccgt 1020 ctggactcac ctcactgtct ccttcgggga gacttcgaca ggttccgaca cgtgttccgt 1080 cacgactggt aactactctt cccgtgtctc cgacgacccc ggtacaaaga ccttcggtag 1140 gggtacaggt aggggggtct tcacttcaag ttgttcggga aacacaagga ctactaactc 1200 gtcttgtggt tctcggggga caagtacccg ttccaacact tggggtgggt ctttact 1257 SEQ ID NO: 8 Exemplified AAT polypeptide Ala Glu Asp Pro Gln Gly Asp Ala Ala Gln Lys Thr Asp Thr Ser His 1 5 10 15 His Asp Gln Asp His Pro Thr Phe Ala Glu Asp Pro Gln Gly Asp Ala 20 25 30 Ala Gln Lys Thr Asp Thr Ser His His Asp Gln Asp His Pro Thr Phe 35 40 45 Asn Lys Ile Thr Pro Asn Leu Ala Glu Phe Ala Phe Ser Leu Tyr Arg 50 55 60 Gln Leu Ala His Gln Ser Asn Ser Thr Asn Ile Phe Phe Ser Pro Val 65 70 75 80 Ser Ile Ala Thr Ala Phe Ala Met Leu Ser Leu Gly Thr Lys Ala Asp 85 90 95 Thr His Asp Glu Ile Leu Glu Gly Leu Asn Phe Asn Leu Thr Glu Ile 100 105 110 Pro Glu Ala Gln Ile His Glu Gly Phe Gln Glu Leu Leu Arg Thr Leu 115 120 125 Asn Gln Pro Asp Ser Gln Leu Gln Leu Thr Thr Gly Asn Gly Leu Phe 130 135 140 Leu Ser Glu Gly Leu Lys Leu Val Asp Lys Phe Leu Glu Asp Val Lys 145 150 155 160 Lys Leu Tyr His Ser Glu Ala Phe Thr Val Asn Phe Gly Asp Thr Glu 165 170 175 Glu Ala Lys Lys Gln Ile Asn Asp Tyr Val Glu Lys Gly Thr Gln Gly 180 185 190 Lys Ile Val Asp Leu Val Lys Glu Leu Asp Arg Asp Thr Val Phe Ala 195 200 205 Leu Val Asn Tyr Ile Phe Phe Lys Gly Lys Trp Glu Arg Pro Phe Glu 210 215 220 Val Lys Asp Thr Glu Glu Glu Asp Phe His Val Asp Gln Val Thr Thr 225 230 235 240 Val Lys Val Pro Met Met Lys Arg Leu Gly Met Phe Asn Ile Gln His 245 250 255 Cys Lys Lys Leu Ser Ser Trp Val Leu Leu Met Lys Tyr Leu Gly Asn 260 265 270 Ala Thr Ala Ile Phe Phe Leu Pro Asp Glu Gly Lys Leu Gln His Leu 275 280 285 Glu Asn Glu Leu Thr His Asp Ile Ile Thr Lys Phe Leu Glu Asn Glu 290 295 300 Asp Arg Arg Ser Ala Ser Leu His Leu Pro Lys Leu Ser Ile Thr Gly 305 310 315 320 Thr Tyr Asp Leu Lys Ser Val Leu Gly Gln Leu Gly Ile Thr Lys Val 325 330 335 Phe Ser Asn Gly Ala Asp Leu Ser Gly Val Thr Glu Glu Ala Pro Leu 340 345 350 Lys Leu Ser Lys Ala Val His Lys Ala Val Leu Thr Ile Asp Glu Lys 355 360 365 Gly Thr Glu Ala Ala Gly Ala Met Phe Leu Glu Ala Ile Pro Met Ser 370 375 380 Ile Pro Pro Glu Val Lys Phe Asn Lys Pro Phe Val Phe Leu Met Ile 385 390 395 400 Glu Gln Asn Thr Lys Ser Pro Leu Phe Met Gly Lys Val Val Asn Pro 405 410 415 Thr Gln Lys SEQ ID NO: 9 Exemplified FVIII transgene (N6) atgcagattg agctgagcac ctgcttcttc ctgtgcctgc tgaggttctg cttctctgcc 60 accaggagat actacctggg ggctgtggag ctgagctggg actacatgca gtctgacctg 120 ggggagctgc ctgtggatgc caggttcccc cccagagtgc ccaagagctt ccccttcaac 180 acctctgtgg tgtacaagaa gaccctgttt gtggagttca ctgaccacct gttcaacatt 240 gccaagccca ggcccccctg gatgggcctg ctgggcccca ccatccaggc tgaggtgtat 300 gacactgtgg tgatcaccct gaagaacatg gccagccacc ctgtgagcct gcatgctgtg 360 ggggtgagct actggaaggc ctctgagggg gctgagtatg atgaccagac cagccagagg 420 gagaaggagg atgacaaggt gttccctggg ggcagccaca cctatgtgtg gcaggtgctg 480 aaggagaatg gccccatggc ctctgacccc ctgtgcctga cctacagcta cctgagccat 540 gtggacctgg tgaaggacct gaactctggc ctgattgggg ccctgctggt gtgcagggag 600 ggcagcctgg ccaaggagaa gacccagacc ctgcacaagt tcatcctgct gtttgctgtg 660 tttgatgagg gcaagagctg gcactctgaa accaagaaca gcctgatgca ggacagggat 720 gctgcctctg ccagggcctg gcccaagatg cacactgtga atggctatgt gaacaggagc 780 ctgcctggcc tgattggctg ccacaggaag tctgtgtact ggcatgtgat tggcatgggc 840 accacccctg aggtgcacag catcttcctg gagggccaca ccttcctggt caggaaccac 900 aggcaggcca gcctggagat cagccccatc accttcctga ctgcccagac cctgctgatg 960 gacctgggcc agttcctgct gttctgccac atcagcagcc accagcatga tggcatggag 1020 gcctatgtga aggtggacag ctgccctgag gagccccagc tgaggatgaa gaacaatgag 1080 gaggctgagg actatgatga tgacctgact gactctgaga tggatgtggt gaggtttgat 1140 gatgacaaca gccccagctt catccagatc aggtctgtgg ccaagaagca ccccaagacc 1200 tgggtgcact acattgctgc tgaggaggag gactgggact atgcccccct ggtgctggcc 1260 cctgatgaca ggagctacaa gagccagtac ctgaacaatg gcccccagag gattggcagg 1320 aagtacaaga aggtcaggtt catggcctac actgatgaaa ccttcaagac cagggaggcc 1380 atccagcatg agtctggcat cctgggcccc ctgctgtatg gggaggtggg ggacaccctg 1440 ctgatcatct tcaagaacca ggccagcagg ccctacaaca tctaccccca tggcatcact 1500 gatgtgaggc ccctgtacag caggaggctg cccaaggggg tgaagcacct gaaggacttc 1560 cccatcctgc ctggggagat cttcaagtac aagtggactg tgactgtgga ggatggcccc 1620 accaagtctg accccaggtg cctgaccaga tactacagca gctttgtgaa catggagagg 1680 gacctggcct ctggcctgat tggccccctg ctgatctgct acaaggagtc tgtggaccag 1740 aggggcaacc agatcatgtc tgacaagagg aatgtgatcc tgttctctgt gtttgatgag 1800 aacaggagct ggtacctgac tgagaacatc cagaggttcc tgcccaaccc tgctggggtg 1860 cagctggagg accctgagtt ccaggccagc aacatcatgc acagcatcaa tggctatgtg 1920 tttgacagcc tgcagctgtc tgtgtgcctg catgaggtgg cctactggta catcctgagc 1980 attggggccc agactgactt cctgtctgtg ttcttctctg gctacacctt caagcacaag 2040 atggtgtatg aggacaccct gaccctgttc cccttctctg gggagactgt gttcatgagc 2100 atggagaacc ctggcctgtg gattctgggc tgccacaact ctgacttcag gaacaggggc 2160 atgactgccc tgctgaaagt ctccagctgt gacaagaaca ctggggacta ctatgaggac 2220 agctatgagg acatctctgc ctacctgctg agcaagaaca atgccattga gcccaggagc 2280 ttcagccaga acagcaggca ccccagcacc aggcagaagc agttcaatgc caccaccatc 2340 cctgagaatg acatagagaa gacagaccca tggtttgccc accggacccc catgcccaag 2400 atccagaatg tgagcagctc tgacctgctg atgctgctga ggcagagccc caccccccat 2460 ggcctgagcc tgtctgacct gcaggaggcc aagtatgaaa ccttctctga tgaccccagc 2520 cctggggcca ttgacagcaa caacagcctg tctgagatga cccacttcag gccccagctg 2580 caccactctg gggacatggt gttcacccct gagtctggcc tgcagctgag gctgaatgag 2640 aagctgggca ccactgctgc cactgagctg aagaagctgg acttcaaagt ctccagcacc 2700 agcaacaacc tgatcagcac catcccctct gacaacctgg ctgctggcac tgacaacacc 2760 agcagcctgg gcccccccag catgcctgtg cactatgaca gccagctgga caccaccctg 2820 tttggcaaga agagcagccc cctgactgag tctgggggcc ccctgagcct gtctgaggag 2880 aacaatgaca gcaagctgct ggagtctggc ctgatgaaca gccaggagag cagctggggc 2940 aagaatgtga gcagcaggga gatcaccagg accaccctgc agtctgacca ggaggagatt 3000 gactatgatg acaccatctc tgtggagatg aagaaggagg actttgacat ctacgacgag 3060 gacgagaacc agagccccag gagcttccag aagaagacca ggcactactt cattgctgct 3120 gtggagaggc tgtgggacta tggcatgagc agcagccccc atgtgctgag gaacagggcc 3180 cagtctggct ctgtgcccca gttcaagaag gtggtgttcc aggagttcac tgatggcagc 3240 ttcacccagc ccctgtacag aggggagctg aatgagcacc tgggcctgct gggcccctac 3300 atcagggctg aggtggagga caacatcatg gtgaccttca ggaaccaggc cagcaggccc 3360 tacagcttct acagcagcct gatcagctat gaggaggacc agaggcaggg ggctgagccc 3420 aggaagaact ttgtgaagcc caatgaaacc aagacctact tctggaaggt gcagcaccac 3480 atggccccca ccaaggatga gtttgactgc aaggcctggg cctacttctc tgatgtggac 3540 ctggagaagg atgtgcactc tggcctgatt ggccccctgc tggtgtgcca caccaacacc 3600 ctgaaccctg cccatggcag gcaggtgact gtgcaggagt ttgccctgtt cttcaccatc 3660 tttgatgaaa ccaagagctg gtacttcact gagaacatgg agaggaactg cagggccccc 3720 tgcaacatcc agatggagga ccccaccttc aaggagaact acaggttcca tgccatcaat 3780 ggctacatca tggacaccct gcctggcctg gtgatggccc aggaccagag gatcaggtgg 3840 tacctgctga gcatgggcag caatgagaac atccacagca tccacttctc tggccatgtg 3900 ttcactgtga ggaagaagga ggagtacaag atggccctgt acaacctgta ccctggggtg 3960 tttgagactg tggagatgct gcccagcaag gctggcatct ggagggtgga gtgcctgatt 4020 ggggagcacc tgcatgctgg catgagcacc ctgttcctgg tgtacagcaa caagtgccag 4080 acccccctgg gcatggcctc tggccacatc agggacttcc agatcactgc ctctggccag 4140 tatggccagt gggcccccaa gctggccagg ctgcactact ctggcagcat caatgcctgg 4200 agcaccaagg agcccttcag ctggatcaag gtggacctgc tggcccccat gatcatccat 4260 ggcatcaaga cccagggggc caggcagaag ttcagcagcc tgtacatcag ccagttcatc 4320 atcatgtaca gcctggatgg caagaagtgg cagacctaca ggggcaacag cactggcacc 4380 ctgatggtgt tctttggcaa tgtggacagc tctggcatca agcacaacat cttcaacccc 4440 cccatcattg ccagatacat caggctgcac cccacccact acagcatcag gagcaccctg 4500 aggatggagc tgatgggctg tgacctgaac agctgcagca tgcccctggg catggagagc 4560 aaggccatct ctgatgccca gatcactgcc agcagctact tcaccaacat gtttgccacc 4620 tggagcccca gcaaggccag gctgcacctg cagggcagga gcaatgcctg gaggccccag 4680 gtcaacaacc ccaaggagtg gctgcaggtg gacttccaga agaccatgaa ggtgactggg 4740 gtgaccaccc agggggtgaa gagcctgctg accagcatgt atgtgaagga gttcctgatc 4800 agcagcagcc aggatggcca ccagtggacc ctgttcttcc agaatggcaa ggtgaaggtg 4860 ttccagggca accaggacag cttcacccct gtggtgaaca gcctggaccc ccccctgctg 4920 accagatacc tgaggattca cccccagagc tgggtgcacc agattgccct gaggatggag 4980 gtgctgggct gtgaggccca ggacctgtac tga 5013 SEQ ID NO: 10 Exemplified FVIII transgene (V3) atgcagattg agctgagcac ctgcttcttc ctgtgcctgc tgaggttctg cttctctgcc 60 accaggagat actacctggg ggctgtggag ctgagctggg actacatgca gtctgacctg 120 ggggagctgc ctgtggatgc caggttcccc cccagagtgc ccaagagctt ccccttcaac 180 acctctgtgg tgtacaagaa gaccctgttt gtggagttca ctgaccacct gttcaacatt 240 gccaagccca ggcccccctg gatgggcctg ctgggcccca ccatccaggc tgaggtgtat 300 gacactgtgg tgatcaccct gaagaacatg gccagccacc ctgtgagcct gcatgctgtg 360 ggggtgagct actggaaggc ctctgagggg gctgagtatg atgaccagac cagccagagg 420 gagaaggagg atgacaaggt gttccctggg ggcagccaca cctatgtgtg gcaggtgctg 480 aaggagaatg gccccatggc ctctgacccc ctgtgcctga cctacagcta cctgagccat 540 gtggacctgg tgaaggacct gaactctggc ctgattgggg ccctgctggt gtgcagggag 600 ggcagcctgg ccaaggagaa gacccagacc ctgcacaagt tcatcctgct gtttgctgtg 660 tttgatgagg gcaagagctg gcactctgaa accaagaaca gcctgatgca ggacagggat 720 gctgcctctg ccagggcctg gcccaagatg cacactgtga atggctatgt gaacaggagc 780 ctgcctggcc tgattggctg ccacaggaag tctgtgtact ggcatgtgat tggcatgggc 840 accacccctg aggtgcacag catcttcctg gagggccaca ccttcctggt caggaaccac 900 aggcaggcca gcctggagat cagccccatc accttcctga ctgcccagac cctgctgatg 960 gacctgggcc agttcctgct gttctgccac atcagcagcc accagcatga tggcatggag 1020 gcctatgtga aggtggacag ctgccctgag gagccccagc tgaggatgaa gaacaatgag 1080 gaggctgagg actatgatga tgacctgact gactctgaga tggatgtggt gaggtttgat 1140 gatgacaaca gccccagctt catccagatc aggtctgtgg ccaagaagca ccccaagacc 1200 tgggtgcact acattgctgc tgaggaggag gactgggact atgcccccct ggtgctggcc 1260 cctgatgaca ggagctacaa gagccagtac ctgaacaatg gcccccagag gattggcagg 1320 aagtacaaga aggtcaggtt catggcctac actgatgaaa ccttcaagac cagggaggcc 1380 atccagcatg agtctggcat cctgggcccc ctgctgtatg gggaggtggg ggacaccctg 1440 ctgatcatct tcaagaacca ggccagcagg ccctacaaca tctaccccca tggcatcact 1500 gatgtgaggc ccctgtacag caggaggctg cccaaggggg tgaagcacct gaaggacttc 1560 cccatcctgc ctggggagat cttcaagtac aagtggactg tgactgtgga ggatggcccc 1620 accaagtctg accccaggtg cctgaccaga tactacagca gctttgtgaa catggagagg 1680 gacctggcct ctggcctgat tggccccctg ctgatctgct acaaggagtc tgtggaccag 1740 aggggcaacc agatcatgtc tgacaagagg aatgtgatcc tgttctctgt gtttgatgag 1800 aacaggagct ggtacctgac tgagaacatc cagaggttcc tgcccaaccc tgctggggtg 1860 cagctggagg accctgagtt ccaggccagc aacatcatgc acagcatcaa tggctatgtg 1920 tttgacagcc tgcagctgtc tgtgtgcctg catgaggtgg cctactggta catcctgagc 1980 attggggccc agactgactt cctgtctgtg ttcttctctg gctacacctt caagcacaag 2040 atggtgtatg aggacaccct gaccctgttc cccttctctg gggagactgt gttcatgagc 2100 atggagaacc ctggcctgtg gattctgggc tgccacaact ctgacttcag gaacaggggc 2160 atgactgccc tgctgaaagt ctccagctgt gacaagaaca ctggggacta ctatgaggac 2220 agctatgagg acatctctgc ctacctgctg agcaagaaca atgccattga gcccaggagc 2280 ttcagccaga atgccactaa tgtgtctaac aacagcaaca ccagcaatga cagcaatgtg 2340 tctcccccag tgctgaagag gcaccagagg gagatcacca ggaccaccct gcagtctgac 2400 caggaggaga ttgactatga tgacaccatc tctgtggaga tgaagaagga ggactttgac 2460 atctacgacg aggacgagaa ccagagcccc aggagcttcc agaagaagac caggcactac 2520 ttcattgctg ctgtggagag gctgtgggac tatggcatga gcagcagccc ccatgtgctg 2580 aggaacaggg cccagtctgg ctctgtgccc cagttcaaga aggtggtgtt ccaggagttc 2640 actgatggca gcttcaccca gcccctgtac agaggggagc tgaatgagca cctgggcctg 2700 ctgggcccct acatcagggc tgaggtggag gacaacatca tggtgacctt caggaaccag 2760 gccagcaggc cctacagctt ctacagcagc ctgatcagct atgaggagga ccagaggcag 2820 ggggctgagc ccaggaagaa ctttgtgaag cccaatgaaa ccaagaccta cttctggaag 2880 gtgcagcacc acatggcccc caccaaggat gagtttgact gcaaggcctg ggcctacttc 2940 tctgatgtgg acctggagaa ggatgtgcac tctggcctga ttggccccct gctggtgtgc 3000 cacaccaaca ccctgaaccc tgcccatggc aggcaggtga ctgtgcagga gtttgccctg 3060 ttcttcacca tctttgatga aaccaagagc tggtacttca ctgagaacat ggagaggaac 3120 tgcagggccc cctgcaacat ccagatggag gaccccacct tcaaggagaa ctacaggttc 3180 catgccatca atggctacat catggacacc ctgcctggcc tggtgatggc ccaggaccag 3240 aggatcaggt ggtacctgct gagcatgggc agcaatgaga acatccacag catccacttc 3300 tctggccatg tgttcactgt gaggaagaag gaggagtaca agatggccct gtacaacctg 3360 taccctgggg tgtttgagac tgtggagatg ctgcccagca aggctggcat ctggagggtg 3420 gagtgcctga ttggggagca cctgcatgct ggcatgagca ccctgttcct ggtgtacagc 3480 aacaagtgcc agacccccct gggcatggcc tctggccaca tcagggactt ccagatcact 3540 gcctctggcc agtatggcca gtgggccccc aagctggcca ggctgcacta ctctggcagc 3600 atcaatgcct ggagcaccaa ggagcccttc agctggatca aggtggacct gctggccccc 3660 atgatcatcc atggcatcaa gacccagggg gccaggcaga agttcagcag cctgtacatc 3720 agccagttca tcatcatgta cagcctggat ggcaagaagt ggcagaccta caggggcaac 3780 agcactggca ccctgatggt gttctttggc aatgtggaca gctctggcat caagcacaac 3840 atcttcaacc cccccatcat tgccagatac atcaggctgc accccaccca ctacagcatc 3900 aggagcaccc tgaggatgga gctgatgggc tgtgacctga acagctgcag catgcccctg 3960 ggcatggaga gcaaggccat ctctgatgcc cagatcactg ccagcagcta cttcaccaac 4020 atgtttgcca cctggagccc cagcaaggcc aggctgcacc tgcagggcag gagcaatgcc 4080 tggaggcccc aggtcaacaa ccccaaggag tggctgcagg tggacttcca gaagaccatg 4140 aaggtgactg gggtgaccac ccagggggtg aagagcctgc tgaccagcat gtatgtgaag 4200 gagttcctga tcagcagcag ccaggatggc caccagtgga ccctgttctt ccagaatggc 4260 aaggtgaagg tgttccaggg caaccaggac agcttcaccc ctgtggtgaa cagcctggac 4320 ccccccctgc tgaccagata cctgaggatt cacccccaga gctgggtgca ccagattgcc 4380 ctgaggatgg aggtgctggg ctgtgaggcc caggacctgt actga 4425 SEQ ID NO: 11 Complementary strand to the exemplified FVIII transgene (N6) tacgtctaac tcgactcgtg gacgaagaag gacacggacg actccaagac gaagagacgg 60 tggtcctcta tgatggaccc ccgacacctc gactcgaccc tgatgtacgt cagactggac 120 cccctcgacg gacacctacg gtccaagggg gggtctcacg ggttctcgaa ggggaagttg 180 tggagacacc acatgttctt ctgggacaaa cacctcaagt gactggtgga caagttgtaa 240 cggttcgggt ccggggggac ctacccggac gacccggggt ggtaggtccg actccacata 300 ctgtgacacc actagtggga cttcttgtac cggtcggtgg gacactcgga cgtacgacac 360 ccccactcga tgaccttccg gagactcccc cgactcatac tactggtctg gtcggtctcc 420 ctcttcctcc tactgttcca caagggaccc ccgtcggtgt ggatacacac cgtccacgac 480 ttcctcttac cggggtaccg gagactgggg gacacggact ggatgtcgat ggactcggta 540 cacctggacc acttcctgga cttgagaccg gactaacccc gggacgacca cacgtccctc 600 ccgtcggacc ggttcctctt ctgggtctgg gacgtgttca agtaggacga caaacgacac 660 aaactactcc cgttctcgac cgtgagactt tggttcttgt cggactacgt cctgtcccta 720 cgacggagac ggtcccggac cgggttctac gtgtgacact taccgataca cttgtcctcg 780 gacggaccgg actaaccgac ggtgtccttc agacacatga ccgtacacta accgtacccg 840 tggtggggac tccacgtgtc gtagaaggac ctcccggtgt ggaaggacca gtccttggtg 900 tccgtccggt cggacctcta gtcggggtag tggaaggact gacgggtctg ggacgactac 960 ctggacccgg tcaaggacga caagacggtg tagtcgtcgg tggtcgtact accgtacctc 1020 cggatacact tccacctgtc gacgggactc ctcggggtcg actcctactt cttgttactc 1080 ctccgactcc tgatactact actggactga ctgagactct acctacacca ctccaaacta 1140 ctactgttgt cggggtcgaa gtaggtctag tccagacacc ggttcttcgt ggggttctgg 1200 acccacgtga tgtaacgacg actcctcctc ctgaccctga tacgggggga ccacgaccgg 1260 ggactactgt cctcgatgtt ctcggtcatg gacttgttac cgggggtctc ctaaccgtcc 1320 ttcatgttct tccagtccaa gtaccggatg tgactacttt ggaagttctg gtccctccgg 1380 taggtcgtac tcagaccgta ggacccgggg gacgacatac ccctccaccc cctgtgggac 1440 gactagtaga agttcttggt ccggtcgtcc gggatgttgt agatgggggt accgtagtga 1500 ctacactccg gggacatgtc gtcctccgac gggttccccc acttcgtgga cttcctgaag 1560 gggtaggacg gacccctcta gaagttcatg ttcacctgac actgacacct cctaccgggg 1620 tggttcagac tggggtccac ggactggtct atgatgtcgt cgaaacactt gtacctctcc 1680 ctggaccgga gaccggacta accgggggac gactagacga tgttcctcag acacctggtc 1740 tccccgttgg tctagtacag actgttctcc ttacactagg acaagagaca caaactactc 1800 ttgtcctcga ccatggactg actcttgtag gtctccaagg acgggttggg acgaccccac 1860 gtcgacctcc tgggactcaa ggtccggtcg ttgtagtacg tgtcgtagtt accgatacac 1920 aaactgtcgg acgtcgacag acacacggac gtactccacc ggatgaccat gtaggactcg 1980 taaccccggg tctgactgaa ggacagacac aagaagagac cgatgtggaa gttcgtgttc 2040 taccacatac tcctgtggga ctgggacaag gggaagagac ccctctgaca caagtactcg 2100 tacctcttgg gaccggacac ctaagacccg acggtgttga gactgaagtc cttgtccccg 2160 tactgacggg acgactttca gaggtcgaca ctgttcttgt gacccctgat gatactcctg 2220 tcgatactcc tgtagagacg gatggacgac tcgttcttgt tacggtaact cgggtcctcg 2280 aagtcggtct tgtcgtccgt ggggtcgtgg tccgtcttcg tcaagttacg gtggtggtag 2340 ggactcttac tgtatctctt ctgtctgggt accaaacggg tggcctgggg gtacgggttc 2400 taggtcttac actcgtcgag actggacgac tacgacgact ccgtctcggg gtggggggta 2460 ccggactcgg acagactgga cgtcctccgg ttcatacttt ggaagagact actggggtcg 2520 ggaccccggt aactgtcgtt gttgtcggac agactctact gggtgaagtc cggggtcgac 2580 gtggtgagac ccctgtacca caagtgggga ctcagaccgg acgtcgactc cgacttactc 2640 ttcgacccgt ggtgacgacg gtgactcgac ttcttcgacc tgaagtttca gaggtcgtgg 2700 tcgttgttgg actagtcgtg gtaggggaga ctgttggacc gacgaccgtg actgttgtgg 2760 tcgtcggacc cgggggggtc gtacggacac gtgatactgt cggtcgacct gtggtgggac 2820 aaaccgttct tctcgtcggg ggactgactc agacccccgg gggactcgga cagactcctc 2880 ttgttactgt cgttcgacga cctcagaccg gactacttgt cggtcctctc gtcgaccccg 2940 ttcttacact cgtcgtccct ctagtggtcc tggtgggacg tcagactggt cctcctctaa 3000 ctgatactac tgtggtagag acacctctac ttcttcctcc tgaaactgta gatgctgctc 3060 ctgctcttgg tctcggggtc ctcgaaggtc ttcttctggt ccgtgatgaa gtaacgacga 3120 cacctctccg acaccctgat accgtactcg tcgtcggggg tacacgactc cttgtcccgg 3180 gtcagaccga gacacggggt caagttcttc caccacaagg tcctcaagtg actaccgtcg 3240 aagtgggtcg gggacatgtc tcccctcgac ttactcgtgg acccggacga cccggggatg 3300 tagtcccgac tccacctcct gttgtagtac cactggaagt ccttggtccg gtcgtccggg 3360 atgtcgaaga tgtcgtcgga ctagtcgata ctcctcctgg tctccgtccc ccgactcggg 3420 tccttcttga aacacttcgg gttactttgg ttctggatga agaccttcca cgtcgtggtg 3480 taccgggggt ggttcctact caaactgacg ttccggaccc ggatgaagag actacacctg 3540 gacctcttcc tacacgtgag accggactaa ccgggggacg accacacggt gtggttgtgg 3600 gacttgggac gggtaccgtc cgtccactga cacgtcctca aacgggacaa gaagtggtag 3660 aaactacttt ggttctcgac catgaagtga ctcttgtacc tctccttgac gtcccggggg 3720 acgttgtagg tctacctcct ggggtggaag ttcctcttga tgtccaaggt acggtagtta 3780 ccgatgtagt acctgtggga cggaccggac cactaccggg tcctggtctc ctagtccacc 3840 atggacgact cgtacccgtc gttactcttg taggtgtcgt aggtgaagag accggtacac 3900 aagtgacact ccttcttcct cctcatgttc taccgggaca tgttggacat gggaccccac 3960 aaactctgac acctctacga cgggtcgttc cgaccgtaga cctcccacct cacggactaa 4020 cccctcgtgg acgtacgacc gtactcgtgg gacaaggacc acatgtcgtt gttcacggtc 4080 tggggggacc cgtaccggag accggtgtag tccctgaagg tctagtgacg gagaccggtc 4140 ataccggtca cccgggggtt cgaccggtcc gacgtgatga gaccgtcgta gttacggacc 4200 tcgtggttcc tcgggaagtc gacctagttc cacctggacg accgggggta ctagtaggta 4260 ccgtagttct gggtcccccg gtccgtcttc aagtcgtcgg acatgtagtc ggtcaagtag 4320 tagtacatgt cggacctacc gttcttcacc gtctggatgt ccccgttgtc gtgaccgtgg 4380 gactaccaca agaaaccgtt acacctgtcg agaccgtagt tcgtgttgta gaagttgggg 4440 gggtagtaac ggtctatgta gtccgacgtg gggtgggtga tgtcgtagtc ctcgtgggac 4500 tcctacctcg actacccgac actggacttg tcgacgtcgt acggggaccc gtacctctcg 4560 ttccggtaga gactacgggt ctagtgacgg tcgtcgatga agtggttgta caaacggtgg 4620 acctcggggt cgttccggtc cgacgtggac gtcccgtcct cgttacggac ctccggggtc 4680 cagttgttgg ggttcctcac cgacgtccac ctgaaggtct tctggtactt ccactgaccc 4740 cactggtggg tcccccactt ctcggacgac tggtcgtaca tacacttcct caaggactag 4800 tcgtcgtcgg tcctaccggt ggtcacctgg gacaagaagg tcttaccgtt ccacttccac 4860 aaggtcccgt tggtcctgtc gaagtgggga caccacttgt cggacctggg gggggacgac 4920 tggtctatgg actcctaagt gggggtctcg acccacgtgg tctaacggga ctcctacctc 4980 cacgacccga cactccgggt cctggacatg act 5013 SEQ ID NO: 12 Complementary strand to the exemplified FVIII transgene (V3) tacgtctaac tcgactcgtg gacgaagaag gacacggacg actccaagac gaagagacgg 60 tggtcctcta tgatggaccc ccgacacctc gactcgaccc tgatgtacgt cagactggac 120 cccctcgacg gacacctacg gtccaagggg gggtctcacg ggttctcgaa ggggaagttg 180 tggagacacc acatgttctt ctgggacaaa cacctcaagt gactggtgga caagttgtaa 240 cggttcgggt ccggggggac ctacccggac gacccggggt ggtaggtccg actccacata 300 ctgtgacacc actagtggga cttcttgtac cggtcggtgg gacactcgga cgtacgacac 360 ccccactcga tgaccttccg gagactcccc cgactcatac tactggtctg gtcggtctcc 420 ctcttcctcc tactgttcca caagggaccc ccgtcggtgt ggatacacac cgtccacgac 480 ttcctcttac cggggtaccg gagactgggg gacacggact ggatgtcgat ggactcggta 540 cacctggacc acttcctgga cttgagaccg gactaacccc gggacgacca cacgtccctc 600 ccgtcggacc ggttcctctt ctgggtctgg gacgtgttca agtaggacga caaacgacac 660 aaactactcc cgttctcgac cgtgagactt tggttcttgt cggactacgt cctgtcccta 720 cgacggagac ggtcccggac cgggttctac gtgtgacact taccgataca cttgtcctcg 780 gacggaccgg actaaccgac ggtgtccttc agacacatga ccgtacacta accgtacccg 840 tggtggggac tccacgtgtc gtagaaggac ctcccggtgt ggaaggacca gtccttggtg 900 tccgtccggt cggacctcta gtcggggtag tggaaggact gacgggtctg ggacgactac 960 ctggacccgg tcaaggacga caagacggtg tagtcgtcgg tggtcgtact accgtacctc 1020 cggatacact tccacctgtc gacgggactc ctcggggtcg actcctactt cttgttactc 1080 ctccgactcc tgatactact actggactga ctgagactct acctacacca ctccaaacta 1140 ctactgttgt cggggtcgaa gtaggtctag tccagacacc ggttcttcgt ggggttctgg 1200 acccacgtga tgtaacgacg actcctcctc ctgaccctga tacgggggga ccacgaccgg 1260 ggactactgt cctcgatgtt ctcggtcatg gacttgttac cgggggtctc ctaaccgtcc 1320 ttcatgttct tccagtccaa gtaccggatg tgactacttt ggaagttctg gtccctccgg 1380 taggtcgtac tcagaccgta ggacccgggg gacgacatac ccctccaccc cctgtgggac 1440 gactagtaga agttcttggt ccggtcgtcc gggatgttgt agatgggggt accgtagtga 1500 ctacactccg gggacatgtc gtcctccgac gggttccccc acttcgtgga cttcctgaag 1560 gggtaggacg gacccctcta gaagttcatg ttcacctgac actgacacct cctaccgggg 1620 tggttcagac tggggtccac ggactggtct atgatgtcgt cgaaacactt gtacctctcc 1680 ctggaccgga gaccggacta accgggggac gactagacga tgttcctcag acacctggtc 1740 tccccgttgg tctagtacag actgttctcc ttacactagg acaagagaca caaactactc 1800 ttgtcctcga ccatggactg actcttgtag gtctccaagg acgggttggg acgaccccac 1860 gtcgacctcc tgggactcaa ggtccggtcg ttgtagtacg tgtcgtagtt accgatacac 1920 aaactgtcgg acgtcgacag acacacggac gtactccacc ggatgaccat gtaggactcg 1980 taaccccggg tctgactgaa ggacagacac aagaagagac cgatgtggaa gttcgtgttc 2040 taccacatac tcctgtggga ctgggacaag gggaagagac ccctctgaca caagtactcg 2100 tacctcttgg gaccggacac ctaagacccg acggtgttga gactgaagtc cttgtccccg 2160 tactgacggg acgactttca gaggtcgaca ctgttcttgt gacccctgat gatactcctg 2220 tcgatactcc tgtagagacg gatggacgac tcgttcttgt tacggtaact cgggtcctcg 2280 aagtcggtct tacggtgatt acacagattg ttgtcgttgt ggtcgttact gtcgttacac 2340 agagggggtc acgacttctc cgtggtctcc ctctagtggt cctggtggga cgtcagactg 2400 gtcctcctct aactgatact actgtggtag agacacctct acttcttcct cctgaaactg 2460 tagatgctgc tcctgctctt ggtctcgggg tcctcgaagg tcttcttctg gtccgtgatg 2520 aagtaacgac gacacctctc cgacaccctg ataccgtact cgtcgtcggg ggtacacgac 2580 tccttgtccc gggtcagacc gagacacggg gtcaagttct tccaccacaa ggtcctcaag 2640 tgactaccgt cgaagtgggt cggggacatg tctcccctcg acttactcgt ggacccggac 2700 gacccgggga tgtagtcccg actccacctc ctgttgtagt accactggaa gtccttggtc 2760 cggtcgtccg ggatgtcgaa gatgtcgtcg gactagtcga tactcctcct ggtctccgtc 2820 ccccgactcg ggtccttctt gaaacacttc gggttacttt ggttctggat gaagaccttc 2880 cacgtcgtgg tgtaccgggg gtggttccta ctcaaactga cgttccggac ccggatgaag 2940 agactacacc tggacctctt cctacacgtg agaccggact aaccggggga cgaccacacg 3000 gtgtggttgt gggacttggg acgggtaccg tccgtccact gacacgtcct caaacgggac 3060 aagaagtggt agaaactact ttggttctcg accatgaagt gactcttgta cctctccttg 3120 acgtcccggg ggacgttgta ggtctacctc ctggggtgga agttcctctt gatgtccaag 3180 gtacggtagt taccgatgta gtacctgtgg gacggaccgg accactaccg ggtcctggtc 3240 tcctagtcca ccatggacga ctcgtacccg tcgttactct tgtaggtgtc gtaggtgaag 3300 agaccggtac acaagtgaca ctccttcttc ctcctcatgt tctaccggga catgttggac 3360 atgggacccc acaaactctg acacctctac gacgggtcgt tccgaccgta gacctcccac 3420 ctcacggact aacccctcgt ggacgtacga ccgtactcgt gggacaagga ccacatgtcg 3480 ttgttcacgg tctgggggga cccgtaccgg agaccggtgt agtccctgaa ggtctagtga 3540 cggagaccgg tcataccggt cacccggggg ttcgaccggt ccgacgtgat gagaccgtcg 3600 tagttacgga cctcgtggtt cctcgggaag tcgacctagt tccacctgga cgaccggggg 3660 tactagtagg taccgtagtt ctgggtcccc cggtccgtct tcaagtcgtc ggacatgtag 3720 tcggtcaagt agtagtacat gtcggaccta ccgttcttca ccgtctggat gtccccgttg 3780 tcgtgaccgt gggactacca caagaaaccg ttacacctgt cgagaccgta gttcgtgttg 3840 tagaagttgg gggggtagta acggtctatg tagtccgacg tggggtgggt gatgtcgtag 3900 tcctcgtggg actcctacct cgactacccg acactggact tgtcgacgtc gtacggggac 3960 ccgtacctct cgttccggta gagactacgg gtctagtgac ggtcgtcgat gaagtggttg 4020 tacaaacggt ggacctcggg gtcgttccgg tccgacgtgg acgtcccgtc ctcgttacgg 4080 acctccgggg tccagttgtt ggggttcctc accgacgtcc acctgaaggt cttctggtac 4140 ttccactgac cccactggtg ggtcccccac ttctcggacg actggtcgta catacacttc 4200 ctcaaggact agtcgtcgtc ggtcctaccg gtggtcacct gggacaagaa ggtcttaccg 4260 ttccacttcc acaaggtccc gttggtcctg tcgaagtggg gacaccactt gtcggacctg 4320 gggggggacg actggtctat ggactcctaa gtgggggtct cgacccacgt ggtctaacgg 4380 gactcctacc tccacgaccc gacactccgg gtcctggaca tgact 4425 SEQ ID NO: 13 Exemplified FVIII polypeptide (N6) Met Gln Ile Glu Leu Ser Thr Cys Phe Phe Leu Cys Leu Leu Arg Phe 1 5 10 15 Cys Phe Ser Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser 20 25 30 Trp Asp Tyr Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg 35 40 45 Phe Pro Pro Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val 50 55 60 Tyr Lys Lys Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile 65 70 75 80 Ala Lys Pro Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln 85 90 95 Ala Glu Val Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser 100 105 110 His Pro Val Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser 115 120 125 Glu Gly Ala Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp 130 135 140 Asp Lys Val Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu 145 150 155 160 Lys Glu Asn Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser 165 170 175 Tyr Leu Ser His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile 180 185 190 Gly Ala Leu Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr 195 200 205 Gln Thr Leu His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly 210 215 220 Lys Ser Trp His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp 225 230 235 240 Ala Ala Ser Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr 245 250 255 Val Asn Arg Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val 260 265 270 Tyr Trp His Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile 275 280 285 Phe Leu Glu Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser 290 295 300 Leu Glu Ile Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met 305 310 315 320 Asp Leu Gly Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His 325 330 335 Asp Gly Met Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro 340 345 350 Gln Leu Arg Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp 355 360 365 Leu Thr Asp Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser 370 375 380 Pro Ser Phe Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr 385 390 395 400 Trp Val His Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro 405 410 415 Leu Val Leu Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn 420 425 430 Asn Gly Pro Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met 435 440 445 Ala Tyr Thr Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu 450 455 460 Ser Gly Ile Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu 465 470 475 480 Leu Ile Ile Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro 485 490 495 His Gly Ile Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys 500 505 510 Gly Val Lys His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe 515 520 525 Lys Tyr Lys Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp 530 535 540 Pro Arg Cys Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg 545 550 555 560 Asp Leu Ala Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu 565 570 575 Ser Val Asp Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val 580 585 590 Ile Leu Phe Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu 595 600 605 Asn Ile Gln Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp 610 615 620 Pro Glu Phe Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val 625 630 635 640 Phe Asp Ser Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp 645 650 655 Tyr Ile Leu Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe 660 665 670 Ser Gly Tyr Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr 675 680 685 Leu Phe Pro Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro 690 695 700 Gly Leu Trp Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly 705 710 715 720 Met Thr Ala Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp 725 730 735 Tyr Tyr Glu Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys 740 745 750 Asn Asn Ala Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro 755 760 765 Ser Thr Arg Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp 770 775 780 Ile Glu Lys Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys 785 790 795 800 Ile Gln Asn Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser 805 810 815 Pro Thr Pro His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr 820 825 830 Glu Thr Phe Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn 835 840 845 Ser Leu Ser Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly 850 855 860 Asp Met Val Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu 865 870 875 880 Lys Leu Gly Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys 885 890 895 Val Ser Ser Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn 900 905 910 Leu Ala Ala Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met 915 920 925 Pro Val His Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys 930 935 940 Ser Ser Pro Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Glu Glu 945 950 955 960 Asn Asn Asp Ser Lys Leu Leu Glu Ser Gly Leu Met Asn Ser Gln Glu 965 970 975 Ser Ser Trp Gly Lys Asn Val Ser Ser Arg Glu Ile Thr Arg Thr Thr 980 985 990 Leu Gln Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val 995 1000 1005 Glu Met Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn 1010 1015 1020 Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile 1025 1030 1035 Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro 1040 1045 1050 His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val Pro Gln Phe 1055 1060 1065 Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe Thr Gln 1070 1075 1080 Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu Gly 1085 1090 1095 Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe 1100 1105 1110 Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile 1115 1120 1125 Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn 1130 1135 1140 Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln 1145 1150 1155 His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp 1160 1165 1170 Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His Ser Gly 1175 1180 1185 Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn Pro 1190 1195 1200 Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe 1205 1210 1215 Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met 1220 1225 1230 Glu Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro 1235 1240 1245 Thr Phe Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile 1250 1255 1260 Met Asp Thr Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile 1265 1270 1275 Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser 1280 1285 1290 Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys Lys Glu Glu 1295 1300 1305 Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe Glu Thr 1310 1315 1320 Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu Cys 1325 1330 1335 Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu 1340 1345 1350 Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly 1355 1360 1365 His Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln 1370 1375 1380 Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn 1385 1390 1395 Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu 1400 1405 1410 Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg 1415 1420 1425 Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr 1430 1435 1440 Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr 1445 1450 1455 Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile 1460 1465 1470 Lys His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg 1475 1480 1485 Leu His Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu 1490 1495 1500 Leu Met Gly Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met 1505 1510 1515 Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr 1520 1525 1530 Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu 1535 1540 1545 His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val Asn Asn 1550 1555 1560 Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys Val 1565 1570 1575 Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met 1580 1585 1590 Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln 1595 1600 1605 Trp Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly 1610 1615 1620 Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro 1625 1630 1635 Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His 1640 1645 1650 Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp 1655 1660 1665 Leu Tyr 1670 SEQ ID NO: 14 Exemplified FVIII polypeptide (V3) Met Gln Ile Glu Leu Ser Thr Cys Phe Phe Leu Cys Leu Leu Arg Phe 1 5 10 15 Cys Phe Ser Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser 20 25 30 Trp Asp Tyr Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg 35 40 45 Phe Pro Pro Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val 50 55 60 Tyr Lys Lys Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile 65 70 75 80 Ala Lys Pro Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln 85 90 95 Ala Glu Val Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser 100 105 110 His Pro Val Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser 115 120 125 Glu Gly Ala Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp 130 135 140 Asp Lys Val Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu 145 150 155 160 Lys Glu Asn Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser 165 170 175 Tyr Leu Ser His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile 180 185 190 Gly Ala Leu Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr 195 200 205 Gln Thr Leu His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly 210 215 220 Lys Ser Trp His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp 225 230 235 240 Ala Ala Ser Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr 245 250 255 Val Asn Arg Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val 260 265 270 Tyr Trp His Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile 275 280 285 Phe Leu Glu Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser 290 295 300 Leu Glu Ile Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met 305 310 315 320 Asp Leu Gly Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His 325 330 335 Asp Gly Met Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro 340 345 350 Gln Leu Arg Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp 355 360 365 Leu Thr Asp Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser 370 375 380 Pro Ser Phe Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr 385 390 395 400 Trp Val His Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro 405 410 415 Leu Val Leu Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn 420 425 430 Asn Gly Pro Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met 435 440 445 Ala Tyr Thr Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu 450 455 460 Ser Gly Ile Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu 465 470 475 480 Leu Ile Ile Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro 485 490 495 His Gly Ile Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys 500 505 510 Gly Val Lys His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe 515 520 525 Lys Tyr Lys Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp 530 535 540 Pro Arg Cys Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg 545 550 555 560 Asp Leu Ala Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu 565 570 575 Ser Val Asp Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val 580 585 590 Ile Leu Phe Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu 595 600 605 Asn Ile Gln Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp 610 615 620 Pro Glu Phe Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val 625 630 635 640 Phe Asp Ser Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp 645 650 655 Tyr Ile Leu Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe 660 665 670 Ser Gly Tyr Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr 675 680 685 Leu Phe Pro Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro 690 695 700 Gly Leu Trp Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly 705 710 715 720 Met Thr Ala Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp 725 730 735 Tyr Tyr Glu Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys 740 745 750 Asn Asn Ala Ile Glu Pro Arg Ser Phe Ser Gln Asn Ala Thr Asn Val 755 760 765 Ser Asn Asn Ser Asn Thr Ser Asn Asp Ser Asn Val Ser Pro Pro Val 770 775 780 Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln Ser Asp 785 790 795 800 Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys 805 810 815 Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser 820 825 830 Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu 835 840 845 Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala 850 855 860 Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe 865 870 875 880 Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu 885 890 895 His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn 900 905 910 Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr 915 920 925 Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro 930 935 940 Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys 945 950 955 960 Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala 965 970 975 Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His Ser Gly 980 985 990 Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala 995 1000 1005 His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe Thr 1010 1015 1020 Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu 1025 1030 1035 Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr 1040 1045 1050 Phe Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met 1055 1060 1065 Asp Thr Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg 1070 1075 1080 Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile 1085 1090 1095 His Phe Ser Gly His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr 1100 1105 1110 Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe Glu Thr Val 1115 1120 1125 Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu Cys Leu 1130 1135 1140 Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu Val 1145 1150 1155 Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His 1160 1165 1170 Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp 1175 1180 1185 Ala Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala 1190 1195 1200 Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu 1205 1210 1215 Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln 1220 1225 1230 Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser 1235 1240 1245 Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly 1250 1255 1260 Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys 1265 1270 1275 His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu 1280 1285 1290 His Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu 1295 1300 1305 Met Gly Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu 1310 1315 1320 Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe 1325 1330 1335 Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His 1340 1345 1350 Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val Asn Asn Pro 1355 1360 1365 Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys Val Thr 1370 1375 1380 Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met Tyr 1385 1390 1395 Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp 1400 1405 1410 Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn 1415 1420 1425 Gln Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu 1430 1435 1440 Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln 1445 1450 1455 Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu 1460 1465 1470 Tyr SEQ ID NO: 15 Exemplified CFTR transgene (soCFTR2) gctagccac atgcagagaa gccctctgga gaaggcctct gtggtgagca agctgttctt 60 cagctggacc aggcccatcc tgaggaaggg ctacaggcag agactggagc tgtctgacat 120 ctaccagatc ccctctgtgg actctgctga caacctgtct gagaagctgg agagggagtg 180 ggatagagag ctggccagca agaagaaccc caagctgatc aatgccctga ggagatgctt 240 cttctggaga ttcatgttct atggcatctt cctgtacctg ggggaagtga ccaaggctgt 300 gcagcctctg ctgctgggca gaatcattgc cagctatgac cctgacaaca aggaggagag 360 gagcattgcc atctacctgg gcattggcct gtgcctgctg ttcattgtga ggaccctgct 420 gctgcaccct gccatctttg gcctgcacca cattggcatg cagatgagga ttgccatgtt 480 cagcctgatc tacaagaaaa ccctgaagct gtccagcaga gtgctggaca agatcagcat 540 tggccagctg gtgagcctgc tgagcaacaa cctgaacaag tttgatgagg gcctggccct 600 ggcccacttt gtgtggattg cccctctgca ggtggccctg ctgatgggcc tgatttggga 660 gctgctgcag gcctctgcct tttgtggcct gggcttcctg attgtgctgg ccctgtttca 720 ggctggcctg ggcaggatga tgatgaagta cagggaccag agggcaggca agatcagtga 780 gaggctggtg atcacctctg agatgattga gaacatccag tctgtgaagg cctactgttg 840 ggaggaagct atggagaaga tgattgaaaa cctgaggcag acagagctga agctgaccag 900 gaaggctgcc tatgtgagat acttcaacag ctctgccttc ttcttctctg gcttctttgt 960 ggtgttcctg tctgtgctgc cctatgccct gatcaagggg atcatcctga gaaagatttt 1020 caccaccatc agcttctgca ttgtgctgag gatggctgtg accagacagt tcccctgggc 1080 tgtgcagacc tggtatgaca gcctgggggc catcaacaag atccaggact tcctgcagaa 1140 gcaggagtac aagaccctgg agtacaacct gaccaccaca gaagtggtga tggagaatgt 1200 gacagccttc tgggaggagg gctttgggga gctgtttgag aaggccaagc agaacaacaa 1260 caacagaaag accagcaatg gggatgactc cctgttcttc tccaacttct ccctgctggg 1320 cacacctgtg ctgaaggaca tcaacttcaa gattgagagg gggcagctgc tggctgtggc 1380 tggatctaca ggggctggca agaccagcct gctgatgatg atcatggggg agctggagcc 1440 ttctgagggc aagatcaagc actctggcag gatcagcttt tgcagccagt tcagctggat 1500 catgcctggc accatcaagg agaacatcat ctttggagtg agctatgatg agtacagata 1560 caggagtgtg atcaaggcct gccagctgga ggaggacatc agcaagtttg ctgagaagga 1620 caacattgtg ctgggggagg gaggcattac actgtctggg ggccagagag ccagaatcag 1680 cctggccagg gctgtgtaca aggatgctga cctgtacctg ctggactccc cctttggcta 1740 cctggatgtg ctgacagaga aggagatttt tgagagctgt gtgtgcaagc tgatggccaa 1800 caagaccaga atcctggtga ccagcaagat ggagcacctg aagaaggctg acaagatcct 1860 gatcctgcat gagggcagca gctacttcta tgggaccttc tctgagctgc agaacctgca 1920 gcctgacttc agctctaagc tgatgggctg tgacagcttt gaccagttct ctgctgagag 1980 gaggaacagc atcctgacag agaccctgca cagattcagc ctggagggag atgcccctgt 2040 gagctggaca gagaccaaga agcagagctt caagcagaca ggggagtttg gggagaagag 2100 gaagaactcc atcctgaacc ccatcaacag catcaggaag ttcagcattg tgcagaaaac 2160 ccccctgcag atgaatggca ttgaggaaga ttctgatgag cccctggaga ggagactgag 2220 cctggtgcct gattctgagc agggagaggc catcctgcct aggatctctg tgatcagcac 2280 aggccctaca ctgcaggcca gaaggaggca gtctgtgctg aacctgatga cccactctgt 2340 gaaccagggc cagaacatcc acaggaaaac cacagcctcc accaggaaag tgagcctggc 2400 ccctcaggcc aatctgacag agctggacat ctacagcagg aggctgtctc aggagacagg 2460 cctggagatt tctgaggaga tcaatgagga ggacctgaaa gagtgcttct ttgatgacat 2520 ggagagcatc cctgctgtga ccacctggaa cacctacctg agatacatca cagtgcacaa 2580 gagcctgatc tttgtgctga tctggtgcct ggtgatcttc ctggctgaag tggctgcctc 2640 tctggtggtg ctgtggctgc tgggaaacac cccactgcag gacaagggca acagcaccca 2700 cagcaggaac aacagctatg ctgtgatcat cacctccacc tccagctact atgtgttcta 2760 catctatgtg ggagtggctg ataccctgct ggctatgggc ttctttagag gcctgcccct 2820 ggtgcacaca ctgatcacag tgagcaagat cctccaccac aagatgctgc actctgtgct 2880 gcaggctcct atgagcaccc tgaataccct gaaggctggg ggcatcctga acagattctc 2940 caaggatatt gccatcctgg atgacctgct gcctctcacc atctttgact tcatccagct 3000 gctgctgatt gtgattgggg ccattgctgt ggtggcagtg ctgcagccct acatctttgt 3060 ggccacagtg cctgtgattg tggccttcat catgctgagg gcctactttc tgcagacctc 3120 ccagcagctg aagcagctgg agtctgaggg cagaagcccc atcttcaccc acctggtgac 3180 aagcctgaag ggcctgtgga ccctgagagc ctttggcagg cagccctact ttgagaccct 3240 gttccacaag gccctgaacc tgcacacagc caactggttc ctctacctgt ccaccctgag 3300 atggttccag atgagaattg agatgatctt tgtcatcttc ttcattgctg tgaccttcat 3360 cagcattctg accacaggag agggagaggg cagagtgggc attatcctga ccctggccat 3420 gaacatcatg agcacactgc agtgggcagt gaacagcagc attgatgtgg acagcctgat 3480 gaggagtgtg agcagagtgt tcaagttcat tgatatgccc acagagggca agcctaccaa 3540 gagcaccaag ccctacaaga atggccagct gagcaaagtg atgatcattg agaacagcca 3600 tgtgaagaag gatgatatct ggcccagtgg aggccagatg acagtgaagg acctgacagc 3660 caagtacaca gaggggggca atgctatcct ggagaacatc tccttcagca tctcccctgg 3720 ccagagagtg ggactgctgg gaagaacagg ctctggcaag tctaccctgc tgtctgcctt 3780 cctgaggctg ctgaacacag agggagagat ccagattgat ggagtgtcct gggacagcat 3840 cacactgcag cagtggagga aggcctttgg tgtgatcccc cagaaagtgt tcatcttcag 3900 tggcaccttc aggaagaacc tggaccccta tgagcagtgg tctgaccagg agatttggaa 3960 agtggctgat gaagtgggcc tgagaagtgt gattgagcag ttccctggca agctggactt 4020 tgtcctggtg gatgggggct gtgtgctgag ccatggccac aagcagctga tgtgcctggc 4080 cagatcagtg ctgagcaagg ccaagatcct gctgctggat gagccttctg cccacctgga 4140 tcctgtgacc taccagatca tcaggaggac cctcaagcag gcctttgctg actgcacagt 4200 catcctgtgt gagcacagga ttgaggccat gctggagtgc cagcagttcc tggtgattga 4260 ggagaacaaa gtgaggcagt atgacagcat ccagaagctg ctgaatgaga ggagcctgtt 4320 caggcaggcc atcagcccct ctgatagagt gaagctgttc ccccacagga acagctccaa 4380 gtgcaagagc aagccccaga ttgctgccct gaaggaggag acagaggagg aagtgcagga 4440 caccaggctg tgagggccc 4459 SEQ ID NO: 16 Exemplified CFTR polypeptide MQRSPLEKASVVSKLFFSWTRPILRKGYRQRLELSDIYQIPSVDSADNLSEKLEREWDRELASKKNPKLINALRR CFFWRFMFYGIFLYLGEVTKAVQPLLLGRIIASYDPDNKEERSIAIYLGIGLCLLFIVRTLLLHPAIFGLHHIGM QMRIAMFSLIYKKTLKLSSRVLDKISIGQLVSLLSNNLNKFDEGLALAHFVWIAPLQVALLMGLIWELLQASAFC GLGFLIVLALFQAGLGRMMMKYRDQRAGKISERLVITSEMIENIQSVKAYCWEEAMEKMIENLRQTELKLTRKAA YVRYFNSSAFFFSGFFVVFLSVLPYALIKGIILRKIFTTISFCIVLRMAVTRQFPWAVQTWYDSLGAINKIQDFL QKQEYKTLEYNLTTTEVVMENVTAFWEEGFGELFEKAKQNNNNRKTSNGDDSLFFSNFSLLGTPVLKDINFKIER GQLLAVAGSTGAGKTSLLMMIMGELEPSEGKIKHSGRISFCSQFSWIMPGTIKENIIFGVSYDEYRYRSVIKACQ LEEDISKFAEKDNIVLGEGGITLSGGQRARISLARAVYKDADLYLLDSPFGYLDVLTEKEIFESCVCKLMANKTR ILVTSKMEHLKKADKILILHEGSSYFYGTFSELQNLQPDFSSKLMGCDSFDQFSAERRNSILTETLHRFSLEGDA PVSWTETKKQSFKQTGEFGEKRKNSILNPINSIRKFSIVQKTPLQMNGIEEDSDEPLERRLSLVPDSEQGEAILP RISVISTGPTLQARRRQSVLNLMTHSVNQGQNIHRKTTASTRKVSLAPQANLTELDIYSRRLSQETGLEISEEIN EEDLKECFFDDMESIPAVTTWNTYLRYITVHKSLIFVLIWCLVIFLAEVAASLVVLWLLGNTPLQDKGNSTHSRN NSYAVIITSTSSYYVFYIYVGVADTLLAMGFFRGLPLVHTLITVSKILHHKMLHSVLQAPMSTLNTLKAGGILNR FSKDIAILDDLLPLTIFDFIQLLLIVIGAIAVVAVLQPYIFVATVPVIVAFIMLRAYFLQTSQQLKQLESEGRSP IFTHLVTSLKGLWTLRAFGRQPYFETLFHKALNLHTANWFLYLSTLRWFQMRIEMIFVIFFIAVTFISILTTGEG EGRVGIILTLAMNIMSTLQWAVNSSIDVDSLMRSVSRVFKFIDMPTEGKPTKSTKPYKNGQLSKVMIIENSHVKK DDIWPSGGQMTVKDLTAKYTEGGNAILENISFSISPGQRVGLLGRTGSGKSTLLSAFLRLLNTEGEIQIDGVSWD SITLQQWRKAFGVIPQKVFIFSGTFRKNLDPYEQWSDQEIWKVADEVGLRSVIEQFPGKLDFVLVDGGCVLSHGH KQLMCLARSVLSKAKILLLDEPSAHLDPVTYQIIRRTLKQAFADCTVILCEHRIEAMLECQQFLVIEENKVRQYD SIQKLLNERSLFRQAISPSDRVKLFPHRNSSKCKSKPQIAALKEETEEEVQDTRL SEQ ID NO: 17 Exemplified hGM-CSF transgene ATGTGGCTGCAGAGCCTGCTGCTCTTGGGCACTGTGGCCTGCAGCATCTCTGCACCCGCCCGCTCGCCCAGCCCC AGCACGCAGCCCTGGGAGCATGTGAATGCCATCCAGGAGGCCCGGCGTCTCCTGAACCTGAGTAGAGACACTGCT GCTGAGATGAATGAAACAGTAGAAGTCATCTCAGAAATGTTTGACCTCCAGGAGCCGACCTGCCTACAGACCCGC CTGGAGCTGTACAAGCAGGGCCTGCGGGGCAGCCTCACCAAGCTCAAGGGCCCCTTGACCATGATGGCCAGCCAC TACAAGCAGCACTGCCCTCCAACCCCGGAAACTTCCTGTGCAACCCAGATTATCACCTTTGAAAGTTTCAAAGAG AACCTGAAGGACTTTCTGCTTGTCATCCCCTTTGACTGCTGGGAGCCAGTCCAGGAGTGA SEQ ID NO: 18 Exemplified hGM-CSF polypeptide MWLQSLLLLGTVACSISAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTR LELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFDCWEPVQE SEQ ID NO: 19 Exemplified mGM-CSF transgene ATGTGGCTGCAGAACCTGCTGTTCCTGGGCATTGTGGTGTACAGCCTGTCTGCCCCTACAAGATCCCCTATCACA GTGACCAGACCTTGGAAACATGTGGAAGCCATCAAAGAGGCCCTGAATCTGCTGGATGACATGCCTGTGACACTG AATGAAGAGGTGGAAGTGGTGTCCAATGAGTTCAGCTTCAAGAAACTGACCTGTGTGCAGACCAGGCTGAAGATT TTTGAGCAGGGCCTGAGAGGCAACTTCACCAAGCTGAAAGGGGCTCTGAACATGACAGCCAGCTACTACCAGACC TACTGTCCTCCTACACCTGAGACAGACTGTGAAACCCAAGTGACCACCTATGCTGACTTCATTGACAGCCTCAAG ACCTTCCTGACAGACATCCCCTTTGAGTGCAAGAAACCTGGCCAGAAGTGA SEQ ID NO: 20 Exemplified mGM-CSF polypeptide MWLQNLLFLGIVVYSLSAPTRSPITVTRPWKHVEAIKEALNLLDDMPVTLNEEVEVVSNEFSFKKLTCVQTRLKI FEQGLRGNFTKLKGALNMTASYYQTYCPPTPETDCETQVTTYADFIDSLKTFLTDIPFECKKPGQK SEQ ID NO: 21 Exemplified Human DCN (Decorin) transgene ATGAAGGCCACTATCATCCTCCTTCTGCTTGCACAAGTTTCCTGGGCTGGACCGTTTCAACAGAGAGGCTTATTT GACTTTATGCTAGAAGATGAGGCTTCTGGGATAGGCCCAGAAGTTCCTGATGACCGCGACTTCGAGCCCTCCCTA GGCCCAGTGTGCCCCTTCCGCTGTCAATGCCATCTTCGAGTGGTCCAGTGTTCTGATTTGGGTCTGGACAAAGTG CCAAAGGATCTTCCCCCTGACACAACTCTGCTAGACCTGCAAAACAACAAAATAACCGAAATCAAAGATGGAGAC TTTAAGAACCTGAAGAACCTTCACGCATTGATTCTTGTCAACAATAAAATTAGCAAAGTTAGTCCTGGAGCATTT ACACCTTTGGTGAAGTTGGAACGACTTTATCTGTCCAAGAATCAGCTGAAGGAATTGCCAGAAAAAATGCCCAAA ACTCTTCAGGAGCTGCGTGCCCATGAGAATGAGATCACCAAAGTGCGAAAAGTTACTTTCAATGGACTGAACCAG ATGATTGTCATAGAACTGGGCACCAATCCGCTGAAGAGCTCAGGAATTGAAAATGGGGCTTTCCAGGGAATGAAG AAGCTCTCCTACATCCGCATTGCTGATACCAATATCACCAGCATTCCTCAAGGTCTTCCTCCTTCCCTTACGGAA TTACATCTTGATGGCAACAAAATCAGCAGAGTTGATGCAGCTAGCCTGAAAGGACTGAATAATTTGGCTAAGTTG GGATTGAGTTTCAACAGCATCTCTGCTGTTGACAATGGCTCTCTGGCCAACACGCCTCATCTGAGGGAGCTTCAC TTGGACAACAACAAGCTTACCAGAGTACCTGGTGGGCTGGCAGAGCATAAGTACATCCAGGTTGTCTACCTTCAT AACAACAATATCTCTGTAGTTGGATCAAGTGACTTCTGCCCACCTGGACACAACACCAAAAAGGCTTCTTATTCG GGTGTGAGTCTTTTCAGCAACCCGGTCCAGTACTGGGAGATACAGCCATCCACCTTCAGATGTGTCTACGTGCGC TCTGCCATTCAACTCGGAAACTATAAGTAA SEQ ID NO: 22 Exemplified Human Decorin polypeptide MKATIILLLLAQVSWAGPFQQRGLFDFMLEDEASGIGPEVPDDRDFEPSLGPVCPFRCQCHLRVVQCSDLGLDKV PKDLPPDTTLLDLQNNKITEIKDGDFKNLKNLHALILVNNKISKVSPGAFTPLVKLERLYLSKNQLKELPEKMPK TLQELRAHENEITKVRKVTFNGLNQMIVIELGTNPLKSSGIENGAFQGMKKLSYIRIADTNITSIPQGLPPSLTE LHLDGNKISRVDAASLKGLNNLAKLGLSFNSISAVDNGSLANTPHLRELHLDNNKLTRVPGGLAEHKYIQVVYLH NNNISVVGSSDFCPPGHNTKKASYSGVSLFSNPVQYWEIQPSTFRCVYVRSAIQLGNYK SEQ ID NO: 23 Exemplified Human TRIM72 transgene ATGTCGGCTGCGCCCGGCCTCCTGCACCAGGAGCTGTCCTGCCCGCTGTGCCTGCAGCTGTTCGACGCGCCCGTG ACAGCCGAGTGCGGCCACAGTTTCTGCCGCGCCTGCCTAGGCCGCGTGGCCGGGGAGCCGGCGGCGGATGGCACC GTTCTCTGCCCCTGCTGCCAGGCCCCCACGCGGCCGCAGGCACTCAGCACCAACCTGCAGCTGGCGCGCCTGGTG GAGGGGCTGGCCCAGGTGCCGCAGGGCCACTGCGAGGAGCACCTGGACCCGCTGAGCATCTACTGCGAGCAGGAC CGCGCGCTGGTGTGCGGAGTGTGCGCCTCACTCGGCTCGCACCGCGGTCATCGCCTCCTGCCTGCCGCCGAGGCC CACGCACGCCTCAAGACACAGCTGCCACAGCAGAAACTGCAGCTGCAGGAGGCATGCATGCGCAAGGAGAAGAGT GTGGCTGTGCTGGAGCATCAGCTGGTGGAGGTGGAGGAGACAGTGCGTCAGTTCCGGGGGGCCGTGGGGGAGCAG CTGGGCAAGATGCGGGTGTTCCTGGCTGCACTGGAGGGCTCCTTGGACCGCGAGGCAGAGCGTGTACGGGGTGAG GCAGGGGTCGCCTTGCGCCGGGAGCTGGGGAGCCTGAACTCTTACCTGGAGCAGCTGCGGCAGATGGAGAAGGTC CTGGAGGAGGTGGCGGACAAGCCGCAGACTGAGTTCCTCATGAAATACTGCCTGGTGACCAGCAGGCTGCAGAAG ATCCTGGCAGAGTCTCCCCCACCCGCCCGTCTGGACATCCAGCTGCCAATTATCTCAGATGACTTCAAATTCCAG GTGTGGAGGAAGATGTTCCGGGCTCTGATGCCAGCGCTGGAGGAGCTGACCTTTGACCCGAGCTCTGCGCACCCG AGCCTGGTGGTGTCTTCCTCTGGCCGCCGCGTGGAGTGCTCGGAGCAGAAGGCGCCGCCGGCCGGGGAGGACCCG CGCCAGTTCGACAAGGCGGTGGCGGTGGTGGCGCACCAGCAGCTCTCCGAGGGCGAGCACTACTGGGAGGTGGAT GTTGGCGACAAGCCGCGCTGGGCGCTGGGCGTGATCGCGGCCGAGGCCCCCCGCCGCGGGCGCCTGCACGCGGTG CCCTCGCAGGGCCTGTGGCTGCTGGGGCTGCGCGAGGGCAAGATCCTGGAGGCACACGTGGAGGCCAAGGAGCCG CGCGCTCTGCGCAGCCCCGAGAGGCGGCCCACGCGCATTGGCCTTTACCTGAGCTTCGGCGACGGCGTCCTCTCC TTCTACGATGCCAGCGACGCCGACGCGCTCGTGCCGCTTTTTGCCTTCCACGAGCGCCTGCCCAGGCCCGTGTAC CCCTTCTTCGACGTGTGCTGGCACGACAAGGGCAAGAATGCCCAGCCGCTGCTGCTCGTGGGTCCCGAAGGCGCC GAGGCCTGA SEQ ID NO: 24 Exemplified Human TRIM72 polypeptide MSAAPGLLHQELSCPLCLQLFDAPVTAECGHSFCRACLGRVAGEPAADGTVLCPCCQAPTRPQALSTNLQLARLV EGLAQVPQGHCEEHLDPLSIYCEQDRALVCGVCASLGSHRGHRLLPAAEAHARLKTQLPQQKLQLQEACMRKEKS VAVLEHQLVEVEETVRQFRGAVGEQLGKMRVFLAALEGSLDREAERVRGEAGVALRRELGSLNSYLEQLRQMEKV LEEVADKPQTEFLMKYCLVTSRLQKILAESPPPARLDIQLPIISDDFKFQVWRKMFRALMPALEELTFDPSSAHP SLVVSSSGRRVECSEQKAPPAGEDPRQFDKAVAVVAHQQLSEGEHYWEVDVGDKPRWALGVIAAEAPRRGRLHAV PSQGLWLLGLREGKILEAHVEAKEPRALRSPERRPTRIGLYLSFGDGVLSFYDASDADALVPLFAFHERLPRPVY PFFDVCWHDKGKNAQPLLLVGPEGAEA SEQ ID NO: 25 Exemplified Human ABCA3 (ABCA3) transgene ATGGCTGTGCTCAGGCAGCTGGCGCTCCTCCTCTGGAAGAACTACACCCTGCAGAAGCGGAAGGTCCTGGTGACG GTCCTGGAACTCTTCCTGCCATTGCTGTTTTCTGGGATCCTCATCTGGCTCCGCTTGAAGATTCAGTCGGAAAAT GTGCCCAACGCCACCATCTACCCGGGCCAGTCCATCCAGGAGCTGCCTCTGTTCTTCACCTTCCCTCCGCCAGGA GACACCTGGGAGCTTGCCTACATCCCTTCTCACAGTGACGCTGCCAAGACCGTCACTGAGACAGTGCGCAGGGCA CTTGTGATCAACATGCGAGTGCGCGGCTTTCCCTCCGAGAAGGACTTTGAGGACTACATTAGGTACGACAACTGC TCGTCCAGCGTGCTGGCCGCCGTGGTCTTCGAGCACCCCTTCAACCACAGCAAGGAGCCCCTGCCGCTGGCGGTG AAATATCACCTACGGTTCAGTTACACACGGAGAAATTACATGTGGACCCAAACAGGCTCCTTTTTCCTGAAAGAG ACAGAAGGCTGGCACACTACTTCCCTTTTCCCGCTTTTCCCAAACCCAGGACCAAGGGAACCTACATCCCCTGAT GGCGGAGAACCTGGGTACATCCGGGAAGGCTTCCTGGCCGTGCAGCATGCTGTGGACCGGGCCATCATGGAGTAC CATGCCGATGCCGCCACACGCCAGCTGTTCCAGAGACTGACGGTGACCATCAAGAGGTTCCCGTACCCGCCGTTC ATCGCAGACCCCTTCCTCGTGGCCATCCAGTACCAGCTGCCCCTGCTGCTGCTGCTCAGCTTCACCTACACCGCG CTCACCATTGCCCGTGCTGTCGTGCAGGAGAAGGAAAGGAGGCTGAAGGAGTACATGCGCATGATGGGGCTCAGC AGCTGGCTGCACTGGAGTGCCTGGTTCCTCTTGTTCTTCCTCTTCCTCCTCATCGCCGCCTCCTTCATGACCCTG CTCTTCTGTGTCAAGGTGAAGCCAAATGTAGCCGTGCTGTCCCGCAGCGACCCCTCCCTGGTGCTCGCCTTCCTG CTGTGCTTCGCCATCTCTACCATCTCCTTCAGCTTCATGGTCAGCACCTTCTTCAGCAAAGCCAACATGGCAGCA GCCTTCGGAGGCTTCCTCTACTTCTTCACCTACATCCCCTACTTCTTCGTGGCCCCTCGGTACAACTGGATGACT CTGAGCCAGAAGCTCTGCTCCTGCCTCCTGTCTAATGTCGCCATGGCAATGGGAGCCCAGCTCATTGGGAAATTT GAGGCGAAAGGCATGGGCATCCAGTGGCGAGACCTCCTGAGTCCCGTCAACGTGGACGACGACTTCTGCTTCGGG CAGGTGCTGGGGATGCTGCTGCTGGACTCTGTGCTCTATGGCCTGGTGACCTGGTACATGGAGGCCGTCTTCCCA GGGCAGTTCGGCGTGCCTCAGCCCTGGTACTTCTTCATCATGCCCTCCTATTGGTGTGGGAAGCCAAGGGCGGTT GCAGGGAAGGAGGAAGAAGACAGTGACCCCGAGAAAGCACTCAGAAACGAGTACTTTGAAGCCGAGCCAGAGGAC CTGGTGGCGGGGATCAAGATCAAGCACCTGTCCAAGGTGTTCAGGGTGGGAAATAAGGACAGGGCGGCCGTCAGA GACCTGAACCTCAACCTGTACGAGGGACAGATCACCGTCCTGCTGGGCCACAACGGTGCCGGGAAGACCACCACC CTCTCCATGCTCACAGGTCTCTTTCCCCCCACCAGTGGACGGGCATACATCAGCGGGTATGAAATTTCCCAGGAC ATGGTTCAGATCCGGAAGAGCCTGGGCCTGTGCCCGCAGCACGACATCCTGTTTGACAACTTGACAGTCGCAGAG CACCTTTATTTCTACGCCCAGCTGAAGGGCCTGTCACGTCAGAAGTGCCCTGAAGAAGTCAAGCAGATGCTGCAC ATCATCGGCCTGGAGGACAAGTGGAACTCACGGAGCCGCTTCCTGAGCGGGGGCATGAGGCGCAAGCTCTCCATC GGCATCGCCCTCATCGCAGGCTCCAAGGTGCTGATACTGGACGAGCCCACCTCGGGCATGGACGCCATCTCCAGG AGGGCCATCTGGGATCTTCTTCAGCGGCAGAAAAGTGACCGCACCATCGTGCTGACCACCCACTTCATGGACGAG GCTGACCTGCTGGGAGACCGCATCGCCATCATGGCCAAGGGGGAGCTGCAGTGCTGCGGGTCCTCGCTGTTCCTC AAGCAGAAATACGGTGCCGGCTATCACATGACGCTGGTGAAGGAGCCGCACTGCAACCCGGAAGACATCTCCCAG CTGGTCCACCACCACGTGCCCAACGCCACGCTGGAGAGCAGCGCTGGGGCCGAGCTGTCTTTCATCCTTCCCAGA GAGAGCACGCACAGGTTTGAAGGTCTCTTTGCTAAACTGGAGAAGAAGCAGAAAGAGCTGGGCATTGCCAGCTTT GGGGCATCCATCACCACCATGGAGGAAGTCTTCCTTCGGGTCGGGAAGCTGGTGGACAGCAGTATGGACATCCAG GCCATCCAGCTCCCTGCCCTGCAGTACCAGCACGAGAGGCGCGCCAGCGACTGGGCTGTGGACAGCAACCTCTGT GGGGCCATGGACCCCTCCGACGGCATTGGAGCCCTCATCGAGGAGGAGCGCACCGCTGTCAAGCTCAACACTGGG CTCGCCCTGCACTGCCAGCAATTCTGGGCCATGTTCCTGAAGAAGGCCGCATACAGCTGGCGCGAGTGGAAAATG GTGGCGGCACAGGTCCTGGTGCCTCTGACCTGCGTCACCCTGGCCCTCCTGGCCATCAACTACTCCTCGGAGCTC TTCGACGACCCCATGCTGAGGCTGACCTTGGGCGAGTACGGCAGAACCGTCGTGCCCTTCTCAGTTCCCGGGACC TCCCAGCTGGGTCAGCAGCTGTCAGAGCATCTGAAAGACGCACTGCAGGCTGAGGGACAGGAGCCCCGCGAGGTG CTCGGTGACCTGGAGGAGTTCTTGATCTTCAGGGCTTCTGTGGAGGGGGGCGGCTTTAATGAGCGGTGCCTTGTG GCAGCGTCCTTCAGAGATGTGGGAGAGCGCACGGTCGTCAACGCCTTGTTCAACAACCAGGCGTACCACTCTCCA GCCACTGCCCTGGCCGTCGTGGACAACCTTCTGTTCAAGCTGCTGTGCGGGCCTCACGCCTCCATTGTGGTCTCC AACTTCCCCCAGCCCCGGAGCGCCCTGCAGGCTGCCAAGGACCAGTTTAACGAGGGCCGGAAGGGATTCGACATT GCCCTCAACCTGCTCTTCGCCATGGCATTCTTGGCCAGCACGTTCTCCATCCTGGCGGTCAGCGAGAGGGCCGTG CAGGCCAAGCATGTGCAGTTTGTGAGTGGAGTCCACGTGGCCAGTTTCTGGCTCTCTGCTCTGCTGTGGGACCTC ATCTCCTTCCTCATCCCCAGTCTGCTGCTGCTGGTGGTGTTTAAGGCCTTCGACGTGCGTGCCTTCACGCGGGAC GGCCACATGGCTGACACCCTGCTGCTGCTCCTGCTCTACGGCTGGGCCATCATCCCCCTCATGTACCTGATGAAC TTCTTCTTCTTGGGGGCGGCCACTGCCTACACGAGGCTGACCATCTTCAACATCCTGTCAGGCATCGCCACCTTC CTGATGGTCACCATCATGCGCATCCCAGCTGTAAAACTGGAAGAACTTTCCAAAACCCTGGATCACGTGTTCCTG GTGCTGCCCAACCACTGTCTGGGGATGGCAGTCAGCAGTTTCTACGAGAACTACGAGACGCGGAGGTACTGCACC TCCTCCGAGGTCGCCGCCCACTACTGCAAGAAATATAACATCCAGTACCAGGAGAACTTCTATGCCTGGAGCGCC CCGGGGGTCGGCCGGTTTGTGGCCTCCATGGCCGCCTCAGGGTGCGCCTACCTCATCCTGCTCTTCCTCATCGAG ACCAACCTGCTTCAGAGACTCAGGGGCATCCTCTGCGCCCTCCGGAGGAGGCGGACACTGACAGAATTATACACC CGGATGCCTGTGCTTCCTGAGGACCAAGATGTAGCGGACGAGAGGACCCGCATCCTGGCCCCCAGTCCGGACTCC CTGCTCCACACACCTCTGATTATCAAGGAGCTCTCCAAGGTGTACGAGCAGCGGGTGCCCCTCCTGGCCGTGGAC AGGCTCTCCCTCGCGGTGCAGAAAGGGGAGTGCTTCGGCCTGCTGGGCTTCAATGGAGCCGGGAAGACCACGACT TTCAAAATGCTGACCGGGGAGGAGAGCCTCACTTCTGGGGATGCCTTTGTCGGGGGTCACAGAATCAGCTCTGAT GTCGGAAAGGTGCGGCAGCGGATCGGCTACTGCCCGCAGTTTGATGCCTTGCTGGACCACATGACAGGCCGGGAG ATGCTGGTCATGTACGCTCGGCTCCGGGGCATCCCTGAGCGCCACATCGGGGCCTGCGTGGAGAACACTCTGCGG GGCCTGCTGCTGGAGCCACATGCCAACAAGCTGGTCAGGACGTACAGTGGTGGTAACAAGCGGAAGCTGAGCACC GGCATCGCCCTGATCGGAGAGCCTGCTGTCATCTTCCTGGACGAGCCGTCCACTGGCATGGACCCCGTGGCCCGG CGCCTGCTTTGGGACACCGTGGCACGAGCCCGAGAGTCTGGCAAGGCCATCATCATCACCTCCCACAGCATGGAG GAGTGTGAGGCCCTGTGCACCCGGCTGGCCATCATGGTGCAGGGGCAGTTCAAGTGCCTGGGCAGCCCCCAGCAC CTCAAGAGCAAGTTCGGCAGCGGCTACTCCCTGCGGGCCAAGGTGCAGAGTGAAGGGCAACAGGAGGCGCTGGAG GAGTTCAAGGCCTTCGTGGACCTGACCTTTCCAGGCAGCGTCCTGGAAGATGAGCACCAAGGCATGGTCCATTAC CACCTGCCGGGCCGTGACCTCAGCTGGGCGAAGGTTTTCGGTATTCTGGAGAAAGCCAAGGAAAAGTACGGCGTG GACGACTACTCCGTGAGCCAGATCTCGCTGGAACAGGTCTTCCTGAGCTTCGCCCACCTGCAGCCGCCCACCGCA GAGGAGGGGCGATGA SEQ ID NO: 26 Exemplified Human ABCA3 polypeptide MAVLRQLALLLWKNYTLQKRKVLVTVLELFLPLLFSGILIWLRLKIQSENVPNATIYPGQSIQELPLFFTFPPPG DTWELAYIPSHSDAAKTVTETVRRALVINMRVRGFPSEKDFEDYIRYDNCSSSVLAAVVFEHPFNHSKEPLPLAV KYHLRFSYTRRNYMWTQTGSFFLKETEGWHTTSLFPLFPNPGPREPTSPDGGEPGYIREGFLAVQHAVDRAIMEY HADAATRQLFQRLTVTIKRFPYPPFIADPFLVAIQYQLPLLLLLSFTYTALTIARAVVQEKERRLKEYMRMMGLS SWLHWSAWFLLFFLFLLIAASFMTLLFCVKVKPNVAVLSRSDPSLVLAFLLCFAISTISFSFMVSTFFSKANMAA AFGGFLYFFTYIPYFFVAPRYNWMTLSQKLCSCLLSNVAMAMGAQLIGKFEAKGMGIQWRDLLSPVNVDDDFCFG QVLGMLLLDSVLYGLVTWYMEAVFPGQFGVPQPWYFFIMPSYWCGKPRAVAGKEEEDSDPEKALRNEYFEAEPED LVAGIKIKHLSKVFRVGNKDRAAVRDLNLNLYEGQITVLLGHNGAGKTTTLSMLTGLFPPTSGRAYISGYEISQD MVQIRKSLGLCPQHDILFDNLTVAEHLYFYAQLKGLSRQKCPEEVKQMLHIIGLEDKWNSRSRFLSGGMRRKLSI GIALIAGSKVLILDEPTSGMDAISRRAIWDLLQRQKSDRTIVLTTHFMDEADLLGDRIAIMAKGELQCCGSSLFL KQKYGAGYHMTLVKEPHCNPEDISQLVHHHVPNATLESSAGAELSFILPRESTHRFEGLFAKLEKKQKELGIASF GASITTMEEVFLRVGKLVDSSMDIQAIQLPALQYQHERRASDWAVDSNLCGAMDPSDGIGALIEEERTAVKLNTG LALHCQQFWAMFLKKAAYSWREWKMVAAQVLVPLTCVTLALLAINYSSELFDDPMLRLTLGEYGRTVVPFSVPGT SQLGQQLSEHLKDALQAEGQEPREVLGDLEEFLIFRASVEGGGFNERCLVAASFRDVGERTVVNALFNNQAYHSP ATALAVVDNLLFKLLCGPHASIVVSNFPQPRSALQAAKDQFNEGRKGFDIALNLLFAMAFLASTFSILAVSERAV QAKHVQFVSGVHVASFWLSALLWDLISFLIPSLLLLVVFKAFDVRAFTRDGHMADTLLLLLLYGWAIIPLMYLMN FFFLGAATAYTRLTIFNILSGIATFLMVTIMRIPAVKLEELSKTLDHVFLVLPNHCLGMAVSSFYENYETRRYCT SSEVAAHYCKKYNIQYQENFYAWSAPGVGRFVASMAASGCAYLILLFLIETNLLQRLRGILCALRRRRTLTELYT RMPVLPEDQDVADERTRILAPSPDSLLHTPLIIKELSKVYEQRVPLLAVDRLSLAVQKGECFGLLGFNGAGKTTT FKMLTGEESLTSGDAFVGGHRISSDVGKVRQRIGYCPQFDALLDHMTGREMLVMYARLRGIPERHIGACVENTLR GLLLEPHANKLVRTYSGGNKRKLSTGIALIGEPAVIFLDEPSTGMDPVARRLLWDTVARARESGKAIIITSHSME ECEALCTRLAIMVQGQFKCLGSPQHLKSKFGSGYSLRAKVQSEGQQEALEEFKAFVDLTFPGSVLEDEHQGMVHY HLPGRDLSWAKVFGILEKAKEKYGVDDYSVSQISLEQVFLSFAHLQPPTAEEGR SEQ ID NO: 27 Exemplified hCEF promoter agatctgtta cataacttat ggtaaatggc ctgcctggct gactgcccaa tgacccctgc 60 ccaatgatgt caataatgat gtatgttccc atgtaatgcc aatagggact ttccattgat 120 gtcaatgggt ggagtattta tggtaactgc ccacttggca gtacatcaag tgtatcatat 180 gccaagtatg ccccctattg atgtcaatga tggtaaatgg cctgcctggc attatgccca 240 gtacatgacc ttatgggact ttcctacttg gcagtacatc tatgtattag tcattgctat 300 taccatggga attcactagt ggagaagagc atgcttgagg gctgagtgcc cctcagtggg 360 cagagagcac atggcccaca gtccctgaga agttgggggg aggggtgggc aattgaactg 420 gtgcctagag aaggtggggc ttgggtaaac tgggaaagtg atgtggtgta ctggctccac 480 ctttttcccc agggtggggg agaaccatat ataagtgcag tagtctctgt gaacattcaa 540 gcttctgcct tctccctcct gtgagtttgc tagc 574 SEQ ID NO: 28 Exemplified CMV promoter ccgcggagat ctcaatattg gccattagcc atattattca ttggttatat agcataaatc 60 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt 120 ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta ttaatagtaa 180 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 240 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 300 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 360 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt 420 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac 480 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 540 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 600 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 660 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 720 ataagcagag ctcgtttagt gaaccgtcag atcactagaa gctttattgc ggtagtttat 780 cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa cttaagctgc 840 agaagttggt cgtgaggcac tgggcaggct agc 873 SEQ ID NO: 29 Exemplified EF1a promoter agatccatat ccgcggcaat tttaaaagaa agggaggaat agggggacag acttcagcag 60 agagactaat taatataata acaacacaat tagaaataca acatttacaa accaaaattc 120 aaaaaatttt aaattttaga gccgcggaga tcccgtgagg ctccggtgcc cgtcagtggg 180 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg 240 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc 300 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt 360 ttcgcaacgg gtttgccgcc agaacacagg ctagc 395 SEQ ID NO: 30 Plasmid as defined in Figure 2A (pDNA1 pGM991) GGTACCTCAATATTGGCCATTAGCCATATTATTCATTGGTTATATAGCATAAATCAATATTGGCTATTGGCCATT GCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGGCATTG ATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTAC ATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGT TCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGC AGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTA TGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGT GATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCAT TGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTGCGATCGCCC GCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGCTGGCTTGTAACT CAGTCTCTTACTAGGAGACCAGCTTGAGCCTGGGTGTTCGCTGGTTAGCCTAACCTGGTTGGCCACCAGGGGTAA GGACTCCTTGGCTTAGAAAGCTAATAAACTTGCCTGCATTAGAGCTTATCTGAGTCAAGTGTCCTCATTGACGCC TCACTCTCTTGAACGGGAATCTTCCTTACTGGGTTCTCTCTCTGACCCAGGCGAGAGAAACTCCAGCAGTGGCGC CCGAACAGGGACTTGAGTGAGAGTGTAGGCACGTACAGCTGAGAAGGCGTCGGACGCGAAGGAAGCGCGGGGTGC GACGCGACCAAGAAGGAGACTTGGTGAGTAGGCTTCTCGAGTGCCGGGAAAAAGCTCGAGCCTAGTTAGAGGACT AGGAGAGGCCGTAGCCGTAACTACTCTTGGGCAAGTAGGGCAGGCGGTGGGTACGCAATGGGGGCGGCTACCTCA GCACTAAATAGGAGACAATTAGACCAATTTGAGAAAATACGACTTCGCCCGAACGGAAAGAAAAAGTACCAAATT AAACATTTAATATGGGCAGGCAAGGAGATGGAGCGCTTCGGCCTCCATGAGAGGTTGTTGGAGACAGAGGAGGGG TGTAAAAGAATCATAGAAGTCCTCTACCCCCTAGAACCAACAGGATCGGAGGGCTTAAAAAGTCTGTTCAATCTT GTGTGCGTGCTATATTGCTTGCACAAGGAACAGAAAGTGAAAGACACAGAGGAAGCAGTAGCAACAGTAAGACAA CACTGCCATCTAGTGGAAAAAGAAAAAAGTGCAACAGAGACATCTAGTGGACAAAAGAAAAATGACAAGGGAATA GCAGCGCCACCTGGTGGCAGTCAGAATTTTCCAGCGCAACAACAAGGAAATGCCTGGGTACATGTACCCTTGTCA CCGCGCACCTTAAATGCGTGGGTAAAAGCAGTAGAGGAGAAAAAATTTGGAGCAGAAATAGTACCCATTTTTTTG TTTCAAGCCCTATCGAATTCCCGCGGCAATTTTAAAAGAAAGGGAGGAATAGGGGGACAGACTTCAGCAGAGAGA CTAATTAATATAATAACAACACAATTAGAAATACAACATTTACAAACCAAAATTCAAAAAATTTTAAATTTTAGA GCCGCGGAGATCTGTTACATAACTTATGGTAAATGGCCTGCCTGGCTGACTGCCCAATGACCCCTGCCCAATGAT GTCAATAATGATGTATGTTCCCATGTAATGCCAATAGGGACTTTCCATTGATGTCAATGGGTGGAGTATTTATGG TAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTATGCCCCCTATTGATGTCAATGATGGTAAAT GGCCTGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTATGTATTAGTCA TTGCTATTACCATGGGAATTCACTAGTGGAGAAGAGCATGCTTGAGGGCTGAGTGCCCCTCAGTGGGCAGAGAGC ACATGGCCCACAGTCCCTGAGAAGTTGGGGGGAGGGGTGGGCAATTGAACTGGTGCCTAGAGAAGGTGGGGCTTG GGTAAACTGGGAAAGTGATGTGGTGTACTGGCTCCACCTTTTTCCCCAGGGTGGGGGAGAACCATATATAAGTGC AGTAGTCTCTGTGAACATTCCTGCCTTCTCCCTCCTGTGAGTTTAAGGTAAGTATCAAGGTTACAAGACAGGTTT AAGGAGACCAATAGAAACTGGGCTTGTCGAGACAGAGAAGACTCTTGCGTTTCTGATAGGCACGCGGCCGCCCGT TTGTGCTAGGGTTCTTAGGCTTCTTGGGGGCTGCTGGAACTGCAATGGGAGCAGCGGCGACAGCCCTGACGGTCC AGTCTCAGCATTTGCTTGCTGGGATACTGCAGCAGCAGAAGAATCTGCTGGCGGCTGTGGAGGCTCAACAGCAGA TGTTGAAGCTGACCATTTGGGGTGTTAAAAACCTCAATGCCCGCGTCACAGCCCTTGAGAAGTACCTAGAGGATC AGGCACGACTAAACTCCTGGGGGTGCGCATGGAAACAAGTATGTCATACCACAGTGGAGTGGCCCTGGACAAATC GGACTCCGGATTGGCAAAATATGACTTGGTTGGAGTGGGAAAGACAAATAGCTGATTTGGAAAGCAACATTACGA GACAATTAGTGAAGGCTAGAGAACAAGAGGAAAAGAATCTAGATGCCTATCAGAAGTTAACTAGTTGGTCAGATT TCTGGTCTTGGTTCGATTTCTCAAAATGGCTTAACATTTTAAAAATGGGATTTTTAGTAATAGTAGGAATAATAG GGTTAAGATTACTTTACACAGTATATGGATGTATAGTGAGGGTTAGGCAGGGATATGTTCCTCTATCTCCACAGA TCCATATGCGGCCGCCTATTGGTCTTACTGACATCCACTTTGCCTTTCTCTCCACAGCTAGCCACCATGCCCAGC TCTGTGTCCTGGGGCATTCTGCTGCTGGCTGGCCTGTGCTGTCTGGTGCCTGTGTCCCTGGCTGAGGACCCTCAG GGGGATGCTGCCCAGAAAACAGACACCTCCCACCATGACCAGGACCACCCCACCTTCAACAAGATCACCCCCAAC CTGGCAGAGTTTGCCTTCAGCCTGTACAGACAGCTGGCCCACCAGAGCAACAGCACCAACATCTTTTTCAGCCCT GTGTCCATTGCCACAGCCTTTGCCATGCTGAGCCTGGGCACCAAGGCTGACACCCATGATGAGATCCTGGAAGGC CTGAACTTCAACCTGACAGAGATCCCTGAGGCCCAGATCCATGAGGGCTTCCAGGAACTGCTGAGAACCCTGAAC CAGCCAGACAGCCAGCTGCAGCTGACAACAGGCAATGGGCTGTTCCTGTCTGAGGGCCTGAAGCTGGTGGACAAG TTTCTGGAAGATGTGAAGAAGCTGTACCACTCTGAGGCCTTCACAGTGAACTTTGGGGACACAGAAGAGGCCAAG AAACAGATCAATGACTATGTGGAAAAGGGCACCCAGGGCAAGATTGTGGACCTTGTGAAAGAGCTGGACAGGGAC ACTGTGTTTGCCCTTGTGAACTACATCTTCTTCAAGGGCAAGTGGGAGAGGCCCTTTGAAGTGAAGGACACTGAG GAAGAGGACTTCCATGTGGACCAAGTGACCACAGTGAAGGTGCCAATGATGAAGAGACTGGGGATGTTCAATATC CAGCACTGCAAGAAACTGAGCAGCTGGGTGCTGCTGATGAAGTACCTGGGCAATGCTACAGCCATATTCTTTCTG CCTGATGAGGGCAAGCTGCAGCACCTGGAAAATGAGCTGACCCATGACATCATCACCAAATTTCTGGAAAATGAG GACAGAAGATCTGCCAGCCTGCATCTGCCCAAGCTGAGCATCACAGGCACATATGACCTGAAGTCTGTGCTGGGA CAGCTGGGAATCACCAAGGTGTTCAGCAATGGGGCAGACCTGAGTGGAGTGACAGAGGAAGCCCCTCTGAAGCTG TCCAAGGCTGTGCACAAGGCAGTGCTGACCATTGATGAGAAGGGCACAGAGGCTGCTGGGGCCATGTTTCTGGAA GCCATCCCCATGTCCATCCCCCCAGAAGTGAAGTTCAACAAGCCCTTTGTGTTCCTGATGATTGAGCAGAACACC AAGAGCCCCCTGTTCATGGGCAAGGTTGTGAACCCCACCCAGAAATGAGGGCCCAATCAACCTCTGGATTACAAA ATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCT TTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTAT GAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGG GGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATC GCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAA TCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCT TCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTT CGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCAAGCTTCGCACTTTTTAAAAGAAAAGGGAGGA CTGGATGGGATTTATTACTCCGATAGGACGCTGGCTTGTAACTCAGTCTCTTACTAGGAGACCAGCTTGAGCCTG GGTGTTCGCTGGTTAGCCTAACCTGGTTGGCCACCAGGGGTAAGGACTCCTTGGCTTAGAAAGCTAATAAACTTG CCTGCATTAGAGCTCTTACGCGTCCCGGGCTCGAGATCCGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCT AACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTAT TTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGG CTTTTGCAAAAAGCTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACA AATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGTCCG CTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAA TACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACC GTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAA GTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTC CTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCT CACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGC CCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAG CAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACT ACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTA GCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAA AAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAG GGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAA TCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAGAAAAACTCATCGAGCATCAAATGAAACTGCAATTTAT TCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGT TCCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCC CCTCGTCAAAAATAAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAACAGCT TATGCATTTCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAAC CGTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAA TCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAATATTTTCACCTGAATCAGGATATTCTTCTAATA CCTGGAATGCTGTTTTTCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGA TGGTCGGAAGAGGCATAAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCAACGCTAC CTTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTTCCCATACAATCGATAGATTGTCGCACCTGATTGCC CGACATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTAGAGCAAG ACGTTTCCCGTTGAATATGGCTCATAACACCCCTTGTATTACTGTTTATGTAAGCAGACAGTTTTATTGTTCATG ATGATATATTTTTATCTTGTGCAATGTAACATCAGAGATTTTGAGACACAACAATTGGTCGACGGATCC hCEF Promoter is underlined β-globulin/IgG chimeric intron comprising SIV RRE Intron is italicised SIV RRE is italicised and double-underlined SEQ ID NO: 31 Plasmid as defined in Figure 2B (pDNA1 pGM691) attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 60 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 120 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 180 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 240 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 300 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 360 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc 420 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg 480 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg 540 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 600 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 660 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt 900 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg 960 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg 1020 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1080 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc 1140 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg 1200 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc 1260 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct 1320 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1380 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct 1440 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc 1500 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg 1560 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg 1620 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg 1680 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgctc gagccaccat 1740 gggagctgcc acatctgccc tgaatagacg gcagctggac cagttcgaga agatcagact 1800 gcggcccaac ggcaagaaga agtaccagat caagcacctg atctgggccg gcaaagagat 1860 ggaaagattc ggcctgcacg agcggctgct ggaaaccgag gaaggctgca agagaattat 1920 cgaggtgctg taccctctgg aacctaccgg ctctgagggc ctgaagtccc tgttcaatct 1980 cgtgtgcgtg ctgtactgcc tgcacaaaga acagaaagtg aaggacaccg aagaggccgt 2040 ggccacagtt agacagcact gccacctggt ggaaaaagag aagtccgcca cagagacaag 2100 cagcggccag aagaagaacg acaagggaat tgctgcccct cctggcggca gccagaattt 2160 tcctgctcag cagcagggaa acgcctgggt gcacgttcca ctgagcccta gaacactgaa 2220 tgcctgggtc aaagccgtgg aagagaagaa gtttggcgcc gagatcgtgc ccatgttcca 2280 ggctctgtct gagggctgca ccccttacga catcaaccag atgctgaacg tgctgggaga 2340 tcaccagggc gctctgcaga tcgtgaaaga gatcatcaac gaagaggctg cccagtggga 2400 cgtgacacat ccattgcctg ctggacctct gccagccgga caactgagag atcctagagg 2460 ctctgatatc gccggcacca ccagctctgt gcaagagcag ctggaatgga tctacaccgc 2520 caatcctaga gtggacgtgg gcgccatcta cagaagatgg atcatcctgg gcctgcagaa 2580 atgcgtgaag atgtacaacc ccgtgtccgt gctggacatc agacagggac ccaaagagcc 2640 cttcaaggac tacgtggacc ggttctataa ggccattaga gccgagcagg ccagcggcga 2700 agtgaagcag tggatgacag agagcctgct gatccagaac gccaatccag actgcaaagt 2760 gatcctgaaa ggcctgggca tgcaccccac actggaagag atgctgacag cctgtcaagg 2820 cgttggcggc ccttcttaca aagccaaagt gatggccgag atgatgcaga ccatgcagaa 2880 ccagaacatg gtgcagcaag gcggccctaa gagacagagg cctcctctga gatgctacaa 2940 ctgcggcaag ttcggccaca tgcagagaca gtgtcctgag cctaggaaaa caaaatgtct 3000 aaagtgtgga aaattgggac acctagcaaa agactgcagg ggacaggtga attttttagg 3060 gtatggacgg tggatggggg caaaaccgag aaattttccc gccgctactc ttggagcgga 3120 accgagtgcg cctcctccac cgagcggcac caccccatac gacccagcaa agaagctcct 3180 gcagcaatat gcagagaaag ggaaacaact gagggagcaa aagaggaatc caccggcaat 3240 gaatccggat tggaccgagg gatattcttt gaactccctc tttggagaag accaataaag 3300 accgtgtaca tcgagggcgt gcccatcaag gctctgctgg atacaggcgc cgacgacacc 3360 atcatcaaag agaacgacct gcagctgagc ggcccttgga ggcctaagat cattggagga 3420 atcggcggag gcctgaacgt caaagagtac aacgaccggg aagtgaagat cgaggacaag 3480 atcctgaggg gcacaatcct gctgggcgcc acacctatca acatcatcgg cagaaatctg 3540 ctggcccctg ccggcgctag actggttatg ggacagctct ctgagaagat ccccgtgaca 3600 cccgtgaagc tgaaagaagg cgctagagga ccttgtgtgc gacagtggcc tctgagcaaa 3660 gagaagattg aggccctgca agaaatctgt agccagctgg aacaagaggg caagatcagc 3720 agagttggcg gcgagaacgc ctacaatacc cctatcttct gcatcaagaa aaaggacaag 3780 agccagtggc ggatgctggt ggactttaga gagctgaaca aggctaccca ggacttcttc 3840 gaggtgcagc tgggaattcc tcatcctgcc ggcctgcgga agatgagaca gatcacagtg 3900 ctggatgtgg gcgacgccta ctacagcatc cctctggacc ccaacttcag aaagtacacc 3960 gccttcacaa tccccaccgt gaacaatcaa ggccctggca tcagatacca gttcaactgc 4020 ctgcctcaag gctggaaggg cagccccacc atttttcaga ataccgccgc cagcatcctg 4080 gaagaaatca agagaaacct gcctgctctg accatcgtgc agtacatgga cgatctgtgg 4140 gtcggaagcc aagagaatga gcacacccac gacaagctgg tggaacagct gagaacaaag 4200 ctgcaggcct ggggcctcga aacccctgag aagaaggtgc agaaagaacc tccttacgag 4260 tggatgggct acaagctgtg gcctcacaag tgggagctga gccggattca gctcgaagag 4320 aaggacgagt ggaccgtgaa cgacatccag aaactcgtgg gcaagctgaa ttgggcagcc 4380 cagctgtatc ccggcctgag gaccaagaac atctgcaagc tgatccgggg aaagaagaac 4440 ctgctggaac tggtcacatg gacacctgag gccgaggccg aatatgccga gaatgccgaa 4500 atcctgaaaa ccgagcaaga ggggacctac tacaagcctg gcattccaat cagagctgcc 4560 gtgcagaaac tggaaggcgg ccagtggtcc taccagttta agcaagaagg ccaggtcctg 4620 aaagtgggca agtacaccaa gcagaagaac acccacacca acgagctgag gacactggct 4680 ggcctggtcc agaaaatctg caaagaggcc ctggtcattt ggggcatcct gcctgttctg 4740 gaactgccca ttgagcggga agtgtgggaa cagtggtggg ccgattactg gcaagtgtct 4800 tggatccccg agtgggactt cgtgtctacc cctcctctgc tgaaactgtg gtacaccctg 4860 acaaaagagc ccattcctaa agaggacgtc tactacgttg acggcgcctg caaccggaac 4920 tccaaagaag gcaaggccgg ctacatcagc cagtacggca agcagagagt ggaaaccctg 4980 gaaaacacca ccaaccagca ggccgagctg accgccatta agatggccct ggaagatagc 5040 ggccccaatg tgaacatcgt gaccgactct cagtacgcca tgggaatcct gacagcccag 5100 cctacacaga gcgatagccc tctggttgag cagatcattg ccctgatgat tcagaagcag 5160 caaatctacc tgcagtgggt gcccgctcac aaaggcatcg gcggaaacga agagatcgat 5220 aagctggtgt ccaagggaat cagacgggtg ctgttcctgg aaaagattga agaggcccaa 5280 gaggaacacg agcgctacca caacaactgg aagaatctgg ccgacaccta cggactgccc 5340 cagatcgtgg ccaaagaaat cgtggctatg tgccccaagt gtcagatcaa gggcgaacct 5400 gtgcacggcc aagtggatgc ttctcctggc acatggcaga tggactgtac ccacctggaa 5460 ggcaaagtgg tcatcgtggc tgtgcacgtg gcctccggct ttattgaggc cgaagtgatc 5520 cccagagaga caggcaaaga aaccgccaag ttcctgctga agatcctgtc cagatggccc 5580 atcacacagc tgcacaccga caacggccct aacttcacat ctcaagaggt ggccgccatc 5640 tgttggtggg gaaagattga gcacacaacc ggcattccct acaatccaca gagccagggc 5700 agcatcgagt ccatgaacaa gcagctcaaa gagattatcg gcaagatccg ggacgactgc 5760 cagtacacag aaacagccgt gctgatggcc tgtcacatcc acaacttcaa gcggaaaggc 5820 ggcatcggag gacagacatc tgccgagaga ctgatcaata tcatcaccac tcagctggaa 5880 atccagcacc tccagaccaa gatccagaag attctgaact tccgggtgta ctaccgcgag 5940 ggcagagatc ctgtttggaa aggcccagca cagctgatct ggaaaggcga aggtgccgtg 6000 gtgctgaagg atggctctga tctgaaggtg gtgcccagac ggaaggccaa gattatcaag 6060 gattacgagc ccaaacagcg cgtgggcaat gaaggcgacg ttgagggcac aagaggcagc 6120 gacaattgaa attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg 6180 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg 6240 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg 6300 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc 6360 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg 6420 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg 6480 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt 6540 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat 6600 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc 6660 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 6720 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 6780 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 6840 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc 6900 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 6960 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 7020 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc 7080 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 7140 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtcc 7200 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 7260 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 7320 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 7380 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 7440 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 7500 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 7560 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 7620 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 7680 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 7740 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 7800 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 7860 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 7920 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 7980 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 8040 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 8100 atctaaagta tatatgagta aacttggtct gacagttaga aaaactcatc gagcatcaaa 8160 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc 8220 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg 8280 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata 8340 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaacagc 8400 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca 8460 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga 8520 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc 8580 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt 8640 tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg 8700 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca 8760 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca 8820 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca 8880 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga 8940 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat 9000 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacaattggt 9060 cgac 9064 SEQ ID NO: 32 Plasmid as defined in Figure 2C (pDNA2a pGM297) attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 60 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 120 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 180 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 240 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 300 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 360 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc 420 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg 480 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg 540 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 600 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 660 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt 900 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg 960 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg 1020 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1080 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc 1140 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg 1200 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc 1260 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct 1320 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1380 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct 1440 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc 1500 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg 1560 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg 1620 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg 1680 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgctc gagactagtg 1740 acttggtgag taggcttcga gcctagttag aggactagga gaggccgtag ccgtaactac 1800 tctgggcaag tagggcaggc ggtgggtacg caatgggggc ggctacctca gcactaaata 1860 ggagacaatt agaccaattt gagaaaatac gacttcgccc gaacggaaag aaaaagtacc 1920 aaattaaaca tttaatatgg gcaggcaagg agatggagcg cttcggcctc catgagaggt 1980 tgttggagac agaggagggg tgtaaaagaa tcatagaagt cctctacccc ctagaaccaa 2040 caggatcgga gggcttaaaa agtctgttca atcttgtgtg cgtactatat tgcttgcaca 2100 aggaacagaa agtgaaagac acagaggaag cagtagcaac agtaagacaa cactgccatc 2160 tagtggaaaa agaaaaaagt gcaacagaga catctagtgg acaaaagaaa aatgacaagg 2220 gaatagcagc gccacctggt ggcagtcaga attttccagc gcaacaacaa ggaaatgcct 2280 gggtacatgt acccttgtca ccgcgcacct taaatgcgtg ggtaaaagca gtagaggaga 2340 aaaaatttgg agcagaaata gtacccatgt ttcaagccct atcagaaggc tgcacaccct 2400 atgacattaa tcagatgctt aatgtgctag gagatcatca aggggcatta caaatagtga 2460 aagagatcat taatgaagaa gcagcccagt gggatgtaac acacccacta cccgcaggac 2520 ccctaccagc aggacagctc agggaccctc gcggctcaga tatagcaggg accaccagct 2580 cagtacaaga acagttagaa tggatctata ctgctaaccc ccgggtagat gtaggtgcca 2640 tctaccggag atggattatt ctaggacttc aaaagtgtgt caaaatgtac aacccagtat 2700 cagtcctaga cattaggcag ggacctaaag agcccttcaa ggattatgtg gacagatttt 2760 acaaggcaat tagagcagaa caagcctcag gggaagtgaa acaatggatg acagaatcat 2820 tactcattca aaatgctaat ccagattgta aggtcatcct gaagggccta ggaatgcacc 2880 ccacccttga agaaatgtta acggcttgtc agggggtagg aggcccaagc tacaaagcaa 2940 aagtaatggc agaaatgatg cagaccatgc aaaatcaaaa catggtgcag cagggaggtc 3000 caaaaagaca aagaccccca ctaagatgtt ataattgtgg aaaatttggc catatgcaaa 3060 gacaatgtcc ggaaccaagg aaaacaaaat gtctaaagtg tggaaaattg ggacacctag 3120 caaaagactg caggggacag gtgaattttt tagggtatgg acggtggatg ggggcaaaac 3180 cgagaaattt tcccgccgct actcttggag cggaaccgag tgcgcctcct ccaccgagcg 3240 gcaccacccc atacgaccca gcaaagaagc tcctgcagca atatgcagag aaagggaaac 3300 aactgaggga gcaaaagagg aatccaccgg caatgaatcc ggattggacc gagggatatt 3360 ctttgaactc cctctttgga gaagaccaat aaagacagtg tatatagaag gggtccccat 3420 taaggcactg ctagacacag gggcagatga caccataatt aaagaaaatg atttacaatt 3480 atcaggtcca tggagaccca aaattatagg gggcatagga ggaggcctta atgtaaaaga 3540 atataacgac agggaagtaa aaatagaaga taaaattttg agaggaacaa tattgttagg 3600 agcaactccc attaatataa taggtagaaa tttgctggcc ccggcaggtg cccggttagt 3660 aatgggacaa ttatcagaaa aaattcctgt cacacctgtc aaattgaagg aaggggctcg 3720 gggaccctgt gtaagacaat ggcctctctc taaagagaag attgaagctt tacaggaaat 3780 atgttcccaa ttagagcagg aaggaaaaat cagtagagta ggaggagaaa atgcatacaa 3840 taccccaata ttttgcataa agaagaagga caaatcccag tggaggatgc tagtagactt 3900 tagagagtta aataaggcaa cccaagattt ctttgaagtg caattaggga taccccaccc 3960 agcaggatta agaaagatga gacagataac agttttagat gtaggagacg cctattattc 4020 cataccattg gatccaaatt ttaggaaata tactgctttt actattccca cagtgaataa 4080 tcagggaccc gggattaggt atcaattcaa ctgtctcccg caagggtgga aaggatctcc 4140 tacaatcttc caaaatacag cagcatccat tttggaggag ataaaaagaa acttgccagc 4200 actaaccatt gtacaataca tggatgattt atgggtaggt tctcaagaaa atgaacacac 4260 ccatgacaaa ttagtagaac agttaagaac aaaattacaa gcctggggct tagaaacccc 4320 agaaaagaag gtgcaaaaag aaccacctta tgagtggatg ggatacaaac tttggcctca 4380 caaatgggaa ctaagcagaa tacaactgga ggaaaaagat gaatggactg tcaatgacat 4440 ccagaagtta gttgggaaac taaattgggc agcacaattg tatccaggtc ttaggaccaa 4500 gaatatatgc aagttaatta gaggaaagaa aaatctgtta gagctagtga cttggacacc 4560 tgaggcagaa gctgaatatg cagaaaatgc agagattctt aaaacagaac aggaaggaac 4620 ctattacaaa ccaggaatac ctattagggc agcagtacag aaattggaag gaggacagtg 4680 gagttaccaa ttcaaacaag aaggacaagt cttgaaagta ggaaaataca ccaagcaaaa 4740 gaacacccat acaaatgaac ttcgcacatt agctggttta gtgcagaaga tttgcaaaga 4800 agctctagtt atttggggga tattaccagt tctagaactc ccgatagaaa gagaggtatg 4860 ggaacaatgg tgggcggatt actggcaggt aagctggatt cccgaatggg attttgtcag 4920 caccccacct ttgctcaaac tatggtacac attaacaaaa gaacccatac ccaaggagga 4980 cgtttactat gtagatggag catgcaacag aaattcaaaa gaaggaaaag caggatacat 5040 ctcacaatac ggaaaacaga gagtagaaac attagaaaac actaccaatc agcaagcaga 5100 attaacagct ataaaaatgg ctttggaaga cagtgggcct aatgtgaaca tagtaacaga 5160 ctctcaatat gcaatgggaa ttttgacagc acaacccaca caaagtgatt caccattagt 5220 agagcaaatt atagccttaa tgatacaaaa gcaacaaata tatttgcagt gggtaccagc 5280 acataaagga ataggaggaa atgaggagat agataaatta gtgagtaaag gcattagaag 5340 agttttattc ttagaaaaaa tagaagaagc tcaagaagag catgaaagat atcataataa 5400 ttggaaaaac ctagcagata catatgggct tccacaaata gtagcaaaag agatagtggc 5460 catgtgtcca aaatgtcaga taaagggaga accagtgcat ggacaagtgg atgcctcacc 5520 tggaacatgg cagatggatt gtactcatct agaaggaaaa gtagtcatag ttgcggtcca 5580 tgtagccagt ggattcatag aagcagaagt catacctagg gaaacaggaa aagaaacggc 5640 aaagtttcta ttaaaaatac tgagtagatg gcctataaca cagttacaca cagacaatgg 5700 gcctaacttt acctcccaag aagtggcagc aatatgttgg tggggaaaaa ttgaacatac 5760 aacaggtata ccatataacc cccaatctca aggatcaata gaaagcatga acaaacaatt 5820 aaaagagata attgggaaaa taagagatga ttgccaatat acagagacag cagtactgat 5880 ggcttgccat attcacaatt ttaaaagaaa gggaggaata gggggacaga cttcagcaga 5940 gagactaatt aatataataa caacacaatt agaaatacaa catttacaaa ccaaaattca 6000 aaaaatttta aattttagag tctactacag agaagggaga gaccctgtgt ggaaaggacc 6060 agcacaatta atctggaaag gggaaggagc agtggtcctc aaggacggaa gtgacctaaa 6120 ggttgtacca agaaggaaag ctaaaattat taaggattat gaacccaaac aaagagtggg 6180 taatgagggt gacgtggaag gtaccagggg atctgataac taaatggcag ggaatagtca 6240 gatattggat gagacaaaga aatttgaaat ggaactatta tatgcatcag ctggcggccg 6300 cgaattcact agtgattccc gtttgtgcta gggttcttag gcttcttggg ggctgctgga 6360 actgcaatgg gagcagcggc gacagccctg acggtccagt ctcagcattt gcttgctggg 6420 atactgcagc agcagaagaa tctgctggcg gctgtggagg ctcaacagca gatgttgaag 6480 ctgaccattt ggggtgttaa aaacctcaat gcccgcgtca cagcccttga gaagtaccta 6540 gaggatcagg cacgactaaa ctcctggggg tgcgcatgga aacaagtatg tcataccaca 6600 gtggagtggc cctggacaaa tcggactccg gattggcaaa atatgacttg gttggagtgg 6660 gaaagacaaa tagctgattt ggaaagcaac attacgagac aattagtgaa ggctagagaa 6720 caagaggaaa agaatctaga tgcctatcag aagttaacta gttggtcaga tttctggtct 6780 tggttcgatt tctcaaaatg gcttaacatt ttaaaaatgg gatttttagt aatagtagga 6840 ataatagggt taagattact ttacacagta tatggatgta tagtgagggt taggcaggga 6900 tatgttcctc tatctccaca gatccatatc caatcgaatt cccgcggccg caattcactc 6960 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca 7020 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg 7080 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt 7140 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg 7200 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt 7260 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata 7320 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt 7380 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac 7440 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc 7500 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 7560 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 7620 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 7680 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 7740 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 7800 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt 7860 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa 7920 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 7980 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ccgcttcctc gctcactgac 8040 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 8100 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 8160 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 8220 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 8280 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 8340 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 8400 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 8460 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 8520 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 8580 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 8640 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 8700 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 8760 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 8820 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 8880 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 8940 taaacttggt ctgacagtta gaaaaactca tcgagcatca aatgaaactg caatttattc 9000 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac 9060 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt 9120 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa 9180 tcaccatgag tgacgactga atccggtgag aatggcaaca gcttatgcat ttctttccag 9240 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg 9300 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa 9360 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt 9420 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg 9480 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata 9540 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct 9600 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc 9660 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg 9720 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc 9780 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct 9840 tgtgcaatgt aacatcagag attttgagac acaacaattg gtcgac 9886 SEQ ID NO: 33 Plasmid as defined in Figure 2D (pDNA2b pGM299) tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 60 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 120 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg 180 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 240 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 300 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 360 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 420 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 480 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 540 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 600 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 660 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 720 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat 780 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc 840 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa 900 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac 960 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta 1020 aggctagagt acttaatacg actcactata ggctagcctc gagaattcga ttatgcccct 1080 aggaccagaa gaaagaagat tgcttcgctt gatttggctc ctttacagca ccaatccata 1140 tccaccaagt ggggaaggga cggccagaca acgccgacga gccaggagaa ggtggagaca 1200 acagcaggat caaattagag tcttggtaga aagactccaa gagcaggtgt atgcagttga 1260 ccgcctggct gacgaggctc aacacttggc tatacaacag ttgcctgacc ctcctcattc 1320 agcttagaat cactagtgaa ttcacgcgtg gtacctctag agtcgacccg ggcggccgct 1380 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg 1440 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag 1500 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga 1560 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga 1620 tccgtcgacc aattgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata 1680 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga 1740 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg 1800 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc 1860 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg 1920 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc 1980 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc 2040 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg 2100 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta 2160 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg 2220 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa 2280 tgcataagct gttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg 2340 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa 2400 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt 2460 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc 2520 agtttcattt gatgctcgat gagtttttct aactgtcaga ccaagtttac tcatatatac 2580 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 2640 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 2700 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2760 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2820 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 2880 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2940 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 3000 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 3060 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 3120 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 3180 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 3240 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 3300 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 3360 ttgctcacat ggctcgacag atct 3384 SEQ ID NO: 34 Plasmid as defined in Figure 2E (pDNA3a pGM301) attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 60 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 120 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 180 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 240 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 300 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 360 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc 420 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg 480 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg 540 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 600 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 660 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt 900 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg 960 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg 1020 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1080 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc 1140 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg 1200 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc 1260 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct 1320 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1380 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct 1440 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc 1500 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg 1560 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg 1620 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg 1680 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgat tgccatggca 1740 acatatatcc agagagtaca gtgcatctca acatcactac tggttgttct caccacattg 1800 gtctcgtgtc agattcccag ggataggctc tctaacatag gggtcatagt cgatgaaggg 1860 aaatcactga agatagctgg atcccacgaa tcgaggtaca tagtactgag tctagttccg 1920 ggggtagact ttgagaatgg gtgcggaaca gcccaggtta tccagtacaa gagcctactg 1980 aacaggctgt taatcccatt gagggatgcc ttagatcttc aggaggctct gataactgtc 2040 accaatgata cgacacaaaa tgccggtgct ccccagtcga gattcttcgg tgctgtgatt 2100 ggtactatcg cacttggagt ggcgacatca gcacaaatca ccgcagggat tgcactagcc 2160 gaagcgaggg aggccaaaag agacatagcg ctcatcaaag aatcgatgac aaaaacacac 2220 aagtctatag aactgctgca aaacgctgtg ggggaacaaa ttcttgctct aaagacactc 2280 caggatttcg tgaatgatga gatcaaaccc gcaataagcg aattaggctg tgagactgct 2340 gccttaagac tgggtataaa attgacacag cattactccg agctgttaac tgcgttcggc 2400 tcgaatttcg gaaccatcgg agagaagagc ctcacgctgc aggcgctgtc ttcactttac 2460 tctgctaaca ttactgagat tatgaccaca atcaggacag ggcagtctaa catctatgat 2520 gtcatttata cagaacagat caaaggaacg gtgatagatg tggatctaga gagatacatg 2580 gtcaccctgt ctgtgaagat ccctattctt tctgaagtcc caggtgtgct catacacaag 2640 gcatcatcta tttcttacaa catagacggg gaggaatggt atgtgactgt ccccagccat 2700 atactcagtc gtgcttcttt cttagggggt gcagacataa ccgattgtgt tgagtccaga 2760 ttgacctata tatgccccag ggatcccgca caactgatac ctgacagcca gcaaaagtgt 2820 atcctggggg acacaacaag gtgtcctgtc acaaaagttg tggacagcct tatccccaag 2880 tttgcttttg tgaatggggg cgttgttgct aactgcatag catccacatg tacctgcggg 2940 acaggccgaa gaccaatcag tcaggatcgc tctaaaggtg tagtattcct aacccatgac 3000 aactgtggtc ttataggtgt caatggggta gaattgtatg ctaaccggag agggcacgat 3060 gccacttggg gggtccagaa cttgacagtc ggtcctgcaa ttgctatcag acccgttgat 3120 atttctctca accttgctga tgctacgaat ttcttgcaag actctaaggc tgagcttgag 3180 aaagcacgga aaatcctctc ggaggtaggt agatggtaca actcaagaga gactgtgatt 3240 acgatcatag tagttatggt cgtaatattg gtggtcatta tagtgatcat catcgtgctt 3300 tatagactca gaaggtgaaa tcactagtga attcactcct caggtgcagg ctgcctatca 3360 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc 3420 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata 3480 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag 3540 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc 3600 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta 3660 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta 3720 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt 3780 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc 3840 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc 3900 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 3960 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4020 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc 4080 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 4140 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 4200 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 4260 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4320 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4380 gtatcttatc atgtctgtcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 4440 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 4500 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 4560 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 4620 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 4680 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 4740 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 4800 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 4860 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 4920 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 4980 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5040 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5100 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 5160 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 5220 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 5280 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttaga 5340 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat 5400 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga 5460 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta 5520 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat 5580 ccggtgagaa tggcaacagc ttatgcattt ctttccagac ttgttcaaca ggccagccat 5640 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct 5700 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca 5760 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt 5820 ctaatacctg gaatgctgtt tttccgggga tcgcagtggt gagtaaccat gcatcatcag 5880 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc 5940 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact 6000 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat 6060 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag 6120 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag 6180 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat 6240 tttgagacac aacaattggt cgac 6264 SEQ ID NO: 35 Plasmid as defined in Figure 2F (pDNA3b pGM303) attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 60 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 120 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 180 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 240 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 300 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 360 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc 420 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg 480 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg 540 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 600 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 660 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt 900 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg 960 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg 1020 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1080 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc 1140 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg 1200 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc 1260 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct 1320 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1380 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct 1440 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc 1500 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg 1560 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc 1620 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt 1680 gctgtctcat cattttggca aagaattcct cgagcatgtg gtctgagtta aaaatcagga 1740 gcaacgacgg aggtgaagga ccagaggacg ccaacgaccc ccggggaaag ggggtgcaac 1800 acatccatat ccagccatct ctacctgttt atggacagag ggttagggat ggtgataggg 1860 gcaaacgtga ctcgtactgg tctacttctc ctagtggtag caccacaaaa ccagcatcag 1920 gttgggagag gtcaagtaaa gccgacacat ggttgctgat tctctcattc acccagtggg 1980 ctttgtcaat tgccacagtg atcatctgta tcataatttc tgctagacaa gggtatagta 2040 tgaaagagta ctcaatgact gtagaggcat tgaacatgag cagcagggag gtgaaagagt 2100 cacttaccag tctaataagg caagaggtta tagcaagggc tgtcaacatt cagagctctg 2160 tgcaaaccgg aatcccagtc ttgttgaaca aaaacagcag ggatgtcatc cagatgattg 2220 ataagtcgtg cagcagacaa gagctcactc agcactgtga gagtacgatc gcagtccacc 2280 atgccgatgg aattgcccca cttgagccac atagtttctg gagatgccct gtcggagaac 2340 cgtatcttag ctcagatcct gaaatctcat tgctgcctgg tccgagcttg ttatctggtt 2400 ctacaacgat ctctggatgt gttaggctcc cttcactctc aattggcgag gcaatctatg 2460 cctattcatc aaatctcatt acacaaggtt gtgctgacat agggaaatca tatcaggtcc 2520 tgcagctagg gtacatatca ctcaattcag atatgttccc tgatcttaac cccgtagtgt 2580 cccacactta tgacatcaac gacaatcgga aatcatgctc tgtggtggca accgggacta 2640 ggggttatca gctttgctcc atgccgactg tagacgaaag aaccgactac tctagtgatg 2700 gtattgagga tctggtcctt gatgtcctgg atctcaaagg gagaactaag tctcaccggt 2760 atcgcaacag cgaggtagat cttgatcacc cgttctctgc actatacccc agtgtaggca 2820 acggcattgc aacagaaggc tcattgatat ttcttgggta tggtggacta accacccctc 2880 tgcagggtga tacaaaatgt aggacccaag gatgccaaca ggtgtcgcaa gacacatgca 2940 atgaggctct gaaaattaca tggctaggag ggaaacaggt ggtcagcgtg atcatccagg 3000 tcaatgacta tctctcagag aggccaaaga taagagtcac aaccattcca atcactcaaa 3060 actatctcgg ggcggaaggt agattattaa aattgggtga tcgggtgtac atctatacaa 3120 gatcatcagg ctggcactct caactgcaga taggagtact tgatgtcagc caccctttga 3180 ctatcaactg gacacctcat gaagccttgt ctagaccagg aaataaagag tgcaattggt 3240 acaataagtg tccgaaggaa tgcatatcag gcgtatacac tgatgcttat ccattgtccc 3300 ctgatgcagc taacgtcgct accgtcacgc tatatgccaa tacatcgcgt gtcaacccaa 3360 caatcatgta ttctaacact actaacatta taaatatgtt aaggataaag gatgttcaat 3420 tagaggctgc atataccacg acatcgtgta tcacgcattt tggtaaaggc tactgctttc 3480 acatcatcga gatcaatcag aagagcctga ataccttaca gccgatgctc tttaagacta 3540 gcatccctaa attatgcaag gccgagtctt aagcggccgc gcatgcgaat tcactcctca 3600 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata 3660 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca 3720 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt 3780 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta 3840 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct 3900 ataaagaggt catcagtata tgaaacagcc ccctgctgtc tattccttat tccatagaaa 3960 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac 4020 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact 4080 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg 4140 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4200 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4260 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga 4320 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 4380 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 4440 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 4500 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa 4560 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4620 ttgtccaaac tcatcaatgt atcttatcat gtctgtccgc ttcctcgctc actgactcgc 4680 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4740 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4800 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4860 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4920 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4980 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5040 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5100 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5160 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5220 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5280 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5340 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5400 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5460 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5520 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5580 cttggtctga cagttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat 5640 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac 5700 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa 5760 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac 5820 catgagtgac gactgaatcc ggtgagaatg gcaacagctt atgcatttct ttccagactt 5880 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat 5940 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac 6000 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac 6060 ctgaatcagg atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga 6120 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt 6180 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc 6240 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac 6300 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg 6360 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg 6420 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg 6480 caatgtaaca tcagagattt tgagacacaa caattggtcg ac 6522 SEQ ID NO: 36 Plasmid as defined in Figure 2G (pDNA2a pGM407) ggtacctcaa tattggccat tagccatatt attcattggt tatatagcat aaatcaatat 60 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc 120 atgtccaata tgaccgccat gttggcattg attattgact agttattaat agtaatcaat 180 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa 240 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt 300 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta 360 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt 420 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc 480 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca 540 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat 600 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa 660 caactgcgat cgcccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 720 tatataagca gagctcgctg gcttgtaact cagtctctta ctaggagacc agcttgagcc 780 tgggtgttcg ctggttagcc taacctggtt ggccaccagg ggtaaggact ccttggctta 840 gaaagctaat aaacttgcct gcattagagc ttatctgagt caagtgtcct cattgacgcc 900 tcactctctt gaacgggaat cttccttact gggttctctc tctgacccag gcgagagaaa 960 ctccagcagt ggcgcccgaa cagggacttg agtgagagtg taggcacgta cagctgagaa 1020 ggcgtcggac gcgaaggaag cgcggggtgc gacgcgacca agaaggagac ttggtgagta 1080 ggcttctcga gtgccgggaa aaagctcgag cctagttaga ggactaggag aggccgtagc 1140 cgtaactact cttgggcaag tagggcaggc ggtgggtacg caatgggggc ggctacctca 1200 gcactaaata ggagacaatt agaccaattt gagaaaatac gacttcgccc gaacggaaag 1260 aaaaagtacc aaattaaaca tttaatatgg gcaggcaagg agatggagcg cttcggcctc 1320 catgagaggt tgttggagac agaggagggg tgtaaaagaa tcatagaagt cctctacccc 1380 ctagaaccaa caggatcgga gggcttaaaa agtctgttca atcttgtgtg cgtgctatat 1440 tgcttgcaca aggaacagaa agtgaaagac acagaggaag cagtagcaac agtaagacaa 1500 cactgccatc tagtggaaaa agaaaaaagt gcaacagaga catctagtgg acaaaagaaa 1560 aatgacaagg gaatagcagc gccacctggt ggcagtcaga attttccagc gcaacaacaa 1620 ggaaatgcct gggtacatgt acccttgtca ccgcgcacct taaatgcgtg ggtaaaagca 1680 gtagaggaga aaaaatttgg agcagaaata gtacccattt ttttgtttca agccctatcg 1740 aattcccgtt tgtgctaggg ttcttaggct tcttgggggc tgctggaact gcaatgggag 1800 cagcggcgac agccctgacg gtccagtctc agcatttgct tgctgggata ctgcagcagc 1860 agaagaatct gctggcggct gtggaggctc aacagcagat gttgaagctg accatttggg 1920 gtgttaaaaa cctcaatgcc cgcgtcacag cccttgagaa gtacctagag gatcaggcac 1980 gactaaactc ctgggggtgc gcatggaaac aagtatgtca taccacagtg gagtggccct 2040 ggacaaatcg gactccggat tggcaaaata tgacttggtt ggagtgggaa agacaaatag 2100 ctgatttgga aagcaacatt acgagacaat tagtgaaggc tagagaacaa gaggaaaaga 2160 atctagatgc ctatcagaag ttaactagtt ggtcagattt ctggtcttgg ttcgatttct 2220 caaaatggct taacatttta aaaatgggat ttttagtaat agtaggaata atagggttaa 2280 gattacttta cacagtatat ggatgtatag tgagggttag gcagggatat gttcctctat 2340 ctccacagat ccatatccgc ggcaatttta aaagaaaggg aggaataggg ggacagactt 2400 cagcagagag actaattaat ataataacaa cacaattaga aatacaacat ttacaaacca 2460 aaattcaaaa aattttaaat tttagagccg cggagatctg ttacataact tatggtaaat 2520 ggcctgcctg gctgactgcc caatgacccc tgcccaatga tgtcaataat gatgtatgtt 2580 cccatgtaat gccaataggg actttccatt gatgtcaatg ggtggagtat ttatggtaac 2640 tgcccacttg gcagtacatc aagtgtatca tatgccaagt atgcccccta ttgatgtcaa 2700 tgatggtaaa tggcctgcct ggcattatgc ccagtacatg accttatggg actttcctac 2760 ttggcagtac atctatgtat tagtcattgc tattaccatg ggaattcact agtggagaag 2820 agcatgcttg agggctgagt gcccctcagt gggcagagag cacatggccc acagtccctg 2880 agaagttggg gggaggggtg ggcaattgaa ctggtgccta gagaaggtgg ggcttgggta 2940 aactgggaaa gtgatgtggt gtactggctc cacctttttc cccagggtgg gggagaacca 3000 tatataagtg cagtagtctc tgtgaacatt caagcttctg ccttctccct cctgtgagtt 3060 tgctagccac catgcccagc tctgtgtcct ggggcattct gctgctggct ggcctgtgct 3120 gtctggtgcc tgtgtccctg gctgaggacc ctcaggggga tgctgcccag aaaacagaca 3180 cctcccacca tgaccaggac caccccacct tcaacaagat cacccccaac ctggcagagt 3240 ttgccttcag cctgtacaga cagctggccc accagagcaa cagcaccaac atctttttca 3300 gccctgtgtc cattgccaca gcctttgcca tgctgagcct gggcaccaag gctgacaccc 3360 atgatgagat cctggaaggc ctgaacttca acctgacaga gatccctgag gcccagatcc 3420 atgagggctt ccaggaactg ctgagaaccc tgaaccagcc agacagccag ctgcagctga 3480 caacaggcaa tgggctgttc ctgtctgagg gcctgaagct ggtggacaag tttctggaag 3540 atgtgaagaa gctgtaccac tctgaggcct tcacagtgaa ctttggggac acagaagagg 3600 ccaagaaaca gatcaatgac tatgtggaaa agggcaccca gggcaagatt gtggaccttg 3660 tgaaagagct ggacagggac actgtgtttg cccttgtgaa ctacatcttc ttcaagggca 3720 agtgggagag gccctttgaa gtgaaggaca ctgaggaaga ggacttccat gtggaccaag 3780 tgaccacagt gaaggtgcca atgatgaaga gactggggat gttcaatatc cagcactgca 3840 agaaactgag cagctgggtg ctgctgatga agtacctggg caatgctaca gccatattct 3900 ttctgcctga tgagggcaag ctgcagcacc tggaaaatga gctgacccat gacatcatca 3960 ccaaatttct ggaaaatgag gacagaagat ctgccagcct gcatctgccc aagctgagca 4020 tcacaggcac atatgacctg aagtctgtgc tgggacagct gggaatcacc aaggtgttca 4080 gcaatggggc agacctgagt ggagtgacag aggaagcccc tctgaagctg tccaaggctg 4140 tgcacaaggc agtgctgacc attgatgaga agggcacaga ggctgctggg gccatgtttc 4200 tggaagccat ccccatgtcc atccccccag aagtgaagtt caacaagccc tttgtgttcc 4260 tgatgattga gcagaacacc aagagccccc tgttcatggg caaggttgtg aaccccaccc 4320 agaaatgagg gcccaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 4380 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 4440 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 4500 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 4560 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 4620 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 4680 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct 4740 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 4800 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 4860 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 4920 cgcaagcttc gcacttttta aaagaaaagg gaggactgga tgggatttat tactccgata 4980 ggacgctggc ttgtaactca gtctcttact aggagaccag cttgagcctg ggtgttcgct 5040 ggttagccta acctggttgg ccaccagggg taaggactcc ttggcttaga aagctaataa 5100 acttgcctgc attagagctc ttacgcgtcc cgggctcgag atccgcatct caattagtca 5160 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 5220 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 5280 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 5340 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5400 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5460 tatcttatca tgtctgtccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 5520 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 5580 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 5640 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 5700 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 5760 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 5820 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 5880 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 5940 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 6000 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 6060 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 6120 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 6180 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 6240 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 6300 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 6360 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttagaa 6420 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata 6480 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat 6540 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa 6600 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc 6660 cggtgagaat ggcaacagct tatgcatttc tttccagact tgttcaacag gccagccatt 6720 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg 6780 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa 6840 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc 6900 taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg 6960 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct 7020 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc 7080 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc 7140 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga 7200 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc 7260 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt 7320 ttgagacaca acaattggtc gacggatcc 7349 SEQ ID NO: 37 Codon-optimised SIV gag-pol nucleic acid sequence (from pGM691) atgggagctg ccacatctgc cctgaataga cggcagctgg accagttcga gaagatcaga 60 ctgcggccca acggcaagaa gaagtaccag atcaagcacc tgatctgggc cggcaaagag 120 atggaaagat tcggcctgca cgagcggctg ctggaaaccg aggaaggctg caagagaatt 180 atcgaggtgc tgtaccctct ggaacctacc ggctctgagg gcctgaagtc cctgttcaat 240 ctcgtgtgcg tgctgtactg cctgcacaaa gaacagaaag tgaaggacac cgaagaggcc 300 gtggccacag ttagacagca ctgccacctg gtggaaaaag agaagtccgc cacagagaca 360 agcagcggcc agaagaagaa cgacaaggga attgctgccc ctcctggcgg cagccagaat 420 tttcctgctc agcagcaggg aaacgcctgg gtgcacgttc cactgagccc tagaacactg 480 aatgcctggg tcaaagccgt ggaagagaag aagtttggcg ccgagatcgt gcccatgttc 540 caggctctgt ctgagggctg caccccttac gacatcaacc agatgctgaa cgtgctggga 600 gatcaccagg gcgctctgca gatcgtgaaa gagatcatca acgaagaggc tgcccagtgg 660 gacgtgacac atccattgcc tgctggacct ctgccagccg gacaactgag agatcctaga 720 ggctctgata tcgccggcac caccagctct gtgcaagagc agctggaatg gatctacacc 780 gccaatccta gagtggacgt gggcgccatc tacagaagat ggatcatcct gggcctgcag 840 aaatgcgtga agatgtacaa ccccgtgtcc gtgctggaca tcagacaggg acccaaagag 900 cccttcaagg actacgtgga ccggttctat aaggccatta gagccgagca ggccagcggc 960 gaagtgaagc agtggatgac agagagcctg ctgatccaga acgccaatcc agactgcaaa 1020 gtgatcctga aaggcctggg catgcacccc acactggaag agatgctgac agcctgtcaa 1080 ggcgttggcg gcccttctta caaagccaaa gtgatggccg agatgatgca gaccatgcag 1140 aaccagaaca tggtgcagca aggcggccct aagagacaga ggcctcctct gagatgctac 1200 aactgcggca agttcggcca catgcagaga cagtgtcctg agcctaggaa aacaaaatgt 1260 ctaaagtgtg gaaaattggg acacctagca aaagactgca ggggacaggt gaatttttta 1320 gggtatggac ggtggatggg ggcaaaaccg agaaattttc ccgccgctac tcttggagcg 1380 gaaccgagtg cgcctcctcc accgagcggc accaccccat acgacccagc aaagaagctc 1440 ctgcagcaat atgcagagaa agggaaacaa ctgagggagc aaaagaggaa tccaccggca 1500 atgaatccgg attggaccga gggatattct ttgaactccc tctttggaga agaccaataa 1560 agaccgtgta catcgagggc gtgcccatca aggctctgct ggatacaggc gccgacgaca 1620 ccatcatcaa agagaacgac ctgcagctga gcggcccttg gaggcctaag atcattggag 1680 gaatcggcgg aggcctgaac gtcaaagagt acaacgaccg ggaagtgaag atcgaggaca 1740 agatcctgag gggcacaatc ctgctgggcg ccacacctat caacatcatc ggcagaaatc 1800 tgctggcccc tgccggcgct agactggtta tgggacagct ctctgagaag atccccgtga 1860 cacccgtgaa gctgaaagaa ggcgctagag gaccttgtgt gcgacagtgg cctctgagca 1920 aagagaagat tgaggccctg caagaaatct gtagccagct ggaacaagag ggcaagatca 1980 gcagagttgg cggcgagaac gcctacaata cccctatctt ctgcatcaag aaaaaggaca 2040 agagccagtg gcggatgctg gtggacttta gagagctgaa caaggctacc caggacttct 2100 tcgaggtgca gctgggaatt cctcatcctg ccggcctgcg gaagatgaga cagatcacag 2160 tgctggatgt gggcgacgcc tactacagca tccctctgga ccccaacttc agaaagtaca 2220 ccgccttcac aatccccacc gtgaacaatc aaggccctgg catcagatac cagttcaact 2280 gcctgcctca aggctggaag ggcagcccca ccatttttca gaataccgcc gccagcatcc 2340 tggaagaaat caagagaaac ctgcctgctc tgaccatcgt gcagtacatg gacgatctgt 2400 gggtcggaag ccaagagaat gagcacaccc acgacaagct ggtggaacag ctgagaacaa 2460 agctgcaggc ctggggcctc gaaacccctg agaagaaggt gcagaaagaa cctccttacg 2520 agtggatggg ctacaagctg tggcctcaca agtgggagct gagccggatt cagctcgaag 2580 agaaggacga gtggaccgtg aacgacatcc agaaactcgt gggcaagctg aattgggcag 2640 cccagctgta tcccggcctg aggaccaaga acatctgcaa gctgatccgg ggaaagaaga 2700 acctgctgga actggtcaca tggacacctg aggccgaggc cgaatatgcc gagaatgccg 2760 aaatcctgaa aaccgagcaa gaggggacct actacaagcc tggcattcca atcagagctg 2820 ccgtgcagaa actggaaggc ggccagtggt cctaccagtt taagcaagaa ggccaggtcc 2880 tgaaagtggg caagtacacc aagcagaaga acacccacac caacgagctg aggacactgg 2940 ctggcctggt ccagaaaatc tgcaaagagg ccctggtcat ttggggcatc ctgcctgttc 3000 tggaactgcc cattgagcgg gaagtgtggg aacagtggtg ggccgattac tggcaagtgt 3060 cttggatccc cgagtgggac ttcgtgtcta cccctcctct gctgaaactg tggtacaccc 3120 tgacaaaaga gcccattcct aaagaggacg tctactacgt tgacggcgcc tgcaaccgga 3180 actccaaaga aggcaaggcc ggctacatca gccagtacgg caagcagaga gtggaaaccc 3240 tggaaaacac caccaaccag caggccgagc tgaccgccat taagatggcc ctggaagata 3300 gcggccccaa tgtgaacatc gtgaccgact ctcagtacgc catgggaatc ctgacagccc 3360 agcctacaca gagcgatagc cctctggttg agcagatcat tgccctgatg attcagaagc 3420 agcaaatcta cctgcagtgg gtgcccgctc acaaaggcat cggcggaaac gaagagatcg 3480 ataagctggt gtccaaggga atcagacggg tgctgttcct ggaaaagatt gaagaggccc 3540 aagaggaaca cgagcgctac cacaacaact ggaagaatct ggccgacacc tacggactgc 3600 cccagatcgt ggccaaagaa atcgtggcta tgtgccccaa gtgtcagatc aagggcgaac 3660 ctgtgcacgg ccaagtggat gcttctcctg gcacatggca gatggactgt acccacctgg 3720 aaggcaaagt ggtcatcgtg gctgtgcacg tggcctccgg ctttattgag gccgaagtga 3780 tccccagaga gacaggcaaa gaaaccgcca agttcctgct gaagatcctg tccagatggc 3840 ccatcacaca gctgcacacc gacaacggcc ctaacttcac atctcaagag gtggccgcca 3900 tctgttggtg gggaaagatt gagcacacaa ccggcattcc ctacaatcca cagagccagg 3960 gcagcatcga gtccatgaac aagcagctca aagagattat cggcaagatc cgggacgact 4020 gccagtacac agaaacagcc gtgctgatgg cctgtcacat ccacaacttc aagcggaaag 4080 gcggcatcgg aggacagaca tctgccgaga gactgatcaa tatcatcacc actcagctgg 4140 aaatccagca cctccagacc aagatccaga agattctgaa cttccgggtg tactaccgcg 4200 agggcagaga tcctgtttgg aaaggcccag cacagctgat ctggaaaggc gaaggtgccg 4260 tggtgctgaa ggatggctct gatctgaagg tggtgcccag acggaaggcc aagattatca 4320 aggattacga gcccaaacag cgcgtgggca atgaaggcga cgttgagggc acaagaggca 4380 gcgacaattg a 4391 SEQ ID NO: 38 Exemplary CAG promoter attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 60 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 120 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 180 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 240 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 300 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 360 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc 420 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg 480 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg 540 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 600 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 660 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 720 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 780 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc 840 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt 900 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg 960 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg 1020 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1080 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc 1140 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg 1200 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc 1260 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct 1320 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1380 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct 1440 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc 1500 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg 1560 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg 1620 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg 1680 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgctc gagccacc 1738 SEQ ID NO: 39 Exemplified WPRE component (mWPRE) gggcccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 60 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 120 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 180 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 240 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 300 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 360 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg 420 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 480 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 540 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcaagct 600 SEQ ID NO: 40 Exemplified Human SFTPB transgene ATGCACCAAGCAGGGTACCCAGGCTGCAGAGGTGCCATGGCTGAGTCACACCTGCTGCAGTGGCTGCTGCTGCTG CTGCCCACGCTCTGTGGCCCAGGCACTGCTGCCTGGACCACCTCATCCTTGGCCTGTGCCCAGGGCCCTGAGTTC TGGTGCCAAAGCCTGGAGCAAGCATTGCAGTGCAGAGCCCTAGGGCATTGCCTACAGGAAGTCTGGGGACATGTG GGAGCCGATGACCTATGCCAAGAGTGTGAGGACATCGTCCACATCCTTAACAAGATGGCCAAGGAGGCCATTTTC CAGGACACGATGAGGAAGTTCCTGGAGCAGGAGTGCAACGTCCTCCCCTTGAAGCTGCTCATGCCCCAGTGCAAC CAAGTGCTTGACGACTACTTCCCCCTGGTCATCGACTACTTCCAGAACCAGACTGACTCAAACGGCATCTGTATG CACCTGGGCCTGTGCAAATCCCGGCAGCCAGAGCCAGAGCAGGAGCCAGGGATGTCAGACCCCCTGCCCAAACCT CTGCGGGACCCTCTGCCAGACCCTCTGCTGGACAAGCTCGTCCTCCCTGTGCTGCCCGGGGCCCTCCAGGCGAGG CCTGGGCCTCACACACAGGATCTCTCCGAGCAGCAATTCCCCATTCCTCTCCCCTATTGCTGGCTCTGCAGGGCT CTGATCAAGCGGATCCAAGCCATGATTCCCAAGGGTGCGCTAGCTGTGGCAGTGGCCCAGGTGTGCCGCGTGGTA CCTCTGGTGGCGGGCGGCATCTGCCAGTGCCTGGCTGAGCGCTACTCCGTCATCCTGCTCGACACGCTGCTGGGC CGCATGCTGCCCCAGCTGGTCTGCCGCCTCGTCCTCCGGTGCTCCATGGATGACAGCGCTGGCCCAAGGTCGCCG ACAGGAGAATGGCTGCCGCGAGACTCTGAGTGCCACCTCTGCATGTCCGTGACCACCCAGGCCGGGAACAGCAGC GAGCAGGCCATACCACAGGCAATGCTCCAGGCCTGTGTTGGCTCCTGGCTGGACAGGGAAAAGTGCAAGCAATTT GTGGAGCAGCACACGCCCCAGCTGCTGACCCTGGTGCCCAGGGGCTGGGATGCCCACACCACCTGCCAGGCCCTC GGGGTGTGTGGGACCATGTCCAGCCCTCTCCAGTGTATCCACAGCCCCGACCTTTGA SEQ ID NO: 41 Exemplified Human SFTPB polypeptide MAESHLLQWLLLLLPTLCGPGTAAWTTSSLACAQGPEFWCQSLEQALQCRALGHCLQEVWGHVGADDLCQECEDI VHILNKMAKEAIFQDTMRKFLEQECNVLPLKLLMPQCNQVLDDYFPLVIDYFQNQTDSNGICMHLGLCKSRQPEP EQEPGMSDPLPKPLRDPLPDPLLDKLVLPVLPGALQARPGPHTQDLSEQQFPIPLPYCWLCRALIKRIQAMIPKG ALAVAVAQVCRVVPLVAGGICQCLAERYSVILLDTLLGRMLPQLVCRLVLRCSMDDSAGPRSPTGEWLPRDSECH LCMSVTTQAGNSSEQAIPQAMLQACVGSWLDREKCKQFVEQHTPQLLTLVPRGWDAHTTCQALGVCGTMSSPLQC IHSPDL SEQ ID NO: 42 Exemplified Human ADAMTS13 transgene ATGCACCAGCGTCACCCCCGGGCAAGATGCCCTCCCCTCTGTGTGGCCGGAATCCTTGCCTGTGGCTTTCTCCTG GGCTGCTGGGGACCCTCCCATTTCCAGCAGAGTTGTCTTCAGGCTTTGGAGCCACAGGCCGTGTCTTCTTACTTG AGCCCTGGTGCTCCCTTAAAAGGCCGCCCTCCTTCCCCTGGCTTCCAGAGGCAGAGGCAGAGGCAGAGGCGGGCT GCAGGCGGCATCCTACACCTGGAGCTGCTGGTGGCCGTGGGCCCCGATGTCTTCCAGGCTCACCAGGAGGACACA GAGCGCTATGTGCTCACCAACCTCAACATCGGGGCAGAACTGCTTCGGGACCCGTCCCTGGGGGCTCAGTTTCGG GTGCACCTGGTGAAGATGGTCATTCTGACAGAGCCTGAGGGTGCTCCAAATATCACAGCCAACCTCACCTCGTCC CTGCTGAGCGTCTGTGGGTGGAGCCAGACCATCAACCCTGAGGACGACACGGATCCTGGCCATGCTGACCTGGTC CTCTATATCACTAGGTTTGACCTGGAGTTGCCTGATGGTAACCGGCAGGTGCGGGGCGTCACCCAGCTGGGCGGT GCCTGCTCCCCAACCTGGAGCTGCCTCATTACCGAGGACACTGGCTTCGACCTGGGAGTCACCATTGCCCATGAG ATTGGGCACAGCTTCGGCCTGGAGCACGACGGCGCGCCCGGCAGCGGCTGCGGCCCCAGCGGACACGTGATGGCT TCGGACGGCGCCGCGCCCCGCGCCGGCCTCGCCTGGTCCCCCTGCAGCCGCCGGCAGCTGCTGAGCCTGCTCAGC GCAGGACGGGCGCGCTGCGTGTGGGACCCGCCGCGGCCTCAACCCGGGTCCGCGGGGCACCCGCCGGATGCGCAG CCTGGCCTCTACTACAGCGCCAACGAGCAGTGCCGCGTGGCCTTCGGCCCCAAGGCTGTCGCCTGCACCTTCGCC AGGGAGCACCTGGATATGTGCCAGGCCCTCTCCTGCCACACAGACCCGCTGGACCAAAGCAGCTGCAGCCGCCTC CTCGTTCCTCTCCTGGATGGGACAGAATGTGGCGTGGAGAAGTGGTGCTCCAAGGGTCGCTGCCGCTCCCTGGTG GAGCTGACCCCCATAGCAGCAGTGCATGGGCGCTGGTCTAGCTGGGGTCCCCGAAGTCCTTGCTCCCGCTCCTGC GGAGGAGGTGTGGTCACCAGGAGGCGGCAGTGCAACAACCCCAGACCTGCCTTTGGGGGGCGTGCATGTGTTGGT GCTGACCTCCAGGCCGAGATGTGCAACACTCAGGCCTGCGAGAAGACCCAGCTGGAGTTCATGTCGCAACAGTGC GCCAGGACCGACGGCCAGCCGCTGCGCTCCTCCCCTGGCGGCGCCTCCTTCTACCACTGGGGTGCTGCTGTACCA CACAGCCAAGGGGATGCTCTGTGCAGACACATGTGCCGGGCCATTGGCGAGAGCTTCATCATGAAGCGTGGAGAC AGCTTCCTCGATGGGACCCGGTGTATGCCAAGTGGCCCCCGGGAGGACGGGACCCTGAGCCTGTGTGTGTCGGGC AGCTGCAGGACATTTGGCTGTGATGGTAGGATGGACTCCCAGCAGGTATGGGACAGGTGCCAGGTGTGTGGTGGG GACAACAGCACGTGCAGCCCACGGAAGGGCTCTTTCACAGCTGGCAGAGCGAGAGAATATGTCACGTTTCTGACA GTTACCCCCAACCTGACCAGTGTCTACATTGCCAACCACAGGCCTCTCTTCACACACTTGGCGGTGAGGATCGGA GGGCGCTATGTCGTGGCTGGGAAGATGAGCATCTCCCCTAACACCACCTACCCCTCCCTCCTGGAGGATGGTCGT GTCGAGTACAGAGTGGCCCTCACCGAGGACCGGCTGCCCCGCCTGGAGGAGATCCGCATCTGGGGACCCCTCCAG GAAGATGCTGACATCCAGGTTTACAGGCGGTATGGCGAGGAGTATGGCAACCTCACCCGCCCAGACATCACCTTC ACCTACTTCCAGCCTAAGCCACGGCAGGCCTGGGTGTGGGCCGCTGTGCGTGGGCCCTGCTCGGTGAGCTGTGGG GCAGGGCTGCGCTGGGTAAACTACAGCTGCCTGGACCAGGCCAGGAAGGAGTTGGTGGAGACTGTCCAGTGCCAA GGGAGCCAGCAGCCACCAGCGTGGCCAGAGGCCTGCGTGCTCGAACCCTGCCCTCCCTACTGGGCGGTGGGAGAC TTCGGCCCATGCAGCGCCTCCTGTGGGGGTGGCCTGCGGGAGCGGCCAGTGCGCTGCGTGGAGGCCCAGGGCAGC CTCCTGAAGACATTGCCCCCAGCCCGGTGCAGAGCAGGGGCCCAGCAGCCAGCTGTGGCGCTGGAAACCTGCAAC CCCCAGCCCTGCCCTGCCAGGTGGGAGGTGTCAGAGCCCAGCTCATGCACATCAGCTGGTGGAGCAGGCCTGGCC TTGGAGAACGAGACCTGTGTGCCAGGGGCAGATGGCCTGGAGGCTCCAGTGACTGAGGGGCCTGGCTCCGTAGAT GAGAAGCTGCCTGCCCCTGAGCCCTGTGTCGGGATGTCATGTCCTCCAGGCTGGGGCCATCTGGATGCCACCTCT GCAGGGGAGAAGGCTCCCTCCCCATGGGGCAGCATCAGGACGGGGGCTCAAGCTGCACACGTGTGGACCCCTGCG GCAGGGTCGTGCTCCGTCTCCTGCGGGCGAGGTCTGATGGAGCTGCGTTTCCTGTGCATGGACTCTGCCCTCAGG GTGCCTGTCCAGGAAGAGCTGTGTGGCCTGGCAAGCAAGCCTGGGAGCCGGCGGGAGGTCTGCCAGGCTGTCCCG TGCCCTGCTCGGTGGCAGTACAAGCTGGCGGCCTGCAGCGTGAGCTGTGGGAGAGGGGTCGTGCGGAGGATCCTG TATTGTGCCCGGGCCCATGGGGAGGACGATGGTGAGGAGATCCTGTTGGACACCCAGTGCCAGGGGCTGCCTCGC CCGGAACCCCAGGAGGCCTGCAGCCTGGAGCCCTGCCCACCTAGGTGGAAAGTCATGTCCCTTGGCCCATGTTCG GCCAGCTGTGGCCTTGGCACTGCTAGACGCTCGGTGGCCTGTGTGCAGCTCGACCAAGGCCAGGACGTGGAGGTG GACGAGGCGGCCTGTGCGGCGCTGGTGCGGCCCGAGGCCAGTGTCCCCTGTCTCATTGCCGACTGCACCTACCGC TGGCATGTTGGCACCTGGATGGAGTGCTCTGTTTCCTGTGGGGATGGCATCCAGCGCCGGCGTGACACCTGCCTC GGACCCCAGGCCCAGGCGCCTGTGCCAGCTGATTTCTGCCAGCACTTGCCCAAGCCGGTGACTGTGCGTGGCTGC TGGGCTGGGCCCTGTGTGGGACAGGGTGCCTGTGGCAGGCAGCACCTTGAGCCAACAGGAACCATTGACATGCGA GGCCCAGGGCAGGCAGACTGTGCAGTGGCCATTGGGCGGCCCCTCGGGGAGGTGGTGACCCTCCGCGTCCTTGAG AGTTCTCTCAACTGCAGTGCGGGGGACATGTTGCTGCTTTGGGGCCGGCTCACCTGGAGGAAGATGTGCAGGAAG CTGTTGGACATGACTTTCAGCTCCAAGACCAACACGCTGGTGGTGAGGCAGCGCTGCGGGCGGCCAGGAGGTGGG GTGCTGCTGCGGTATGGGAGCCAGCTTGCTCCTGAAACCTTCTACAGAGAATGTGACATGCAGCTCTTTGGGCCC TGGGGTGAAATCGTGAGCCCCTCGCTGAGTCCAGCCACGAGTAATGCAGGGGGCTGCCGGCTCTTCATTAATGTG GCTCCGCACGCACGGATTGCCATCCATGCCCTGGCCACCAACATGGGCGCTGGGACCGAGGGAGCCAATGCCAGC TACATCTTGATCCGGGACACCCACAGCTTGAGGACCACAGCGTTCCATGGGCAGCAGGTGCTCTACTGGGAGTCA GAGAGCAGCCAGGCTGAGATGGAGTTCAGCGAGGGCTTCCTGAAGGCTCAGGCCAGCCTGCGGGGCCAGTACTGG ACCCTCCAATCATGGGTACCGGAGATGCAGGACCCTCAGTCCTGGAAGGGAAAGGAAGGAACCTGA SEQ ID NO: 43 Exemplified Human ADAMTS13 polypeptide MHQRHPRARCPPLCVAGILACGFLLGCWGPSHFQQSCLQALEPQAVSSYLSPGAPLKGRPPSPGFQRQRQRQRRA AGGILHLELLVAVGPDVFQAHQEDTERYVLTNLNIGAELLRDPSLGAQFRVHLVKMVILTEPEGAPNITANLTSS LLSVCGWSQTINPEDDTDPGHADLVLYITRFDLELPDGNRQVRGVTQLGGACSPTWSCLITEDTGFDLGVTIAHE IGHSFGLEHDGAPGSGCGPSGHVMASDGAAPRAGLAWSPCSRRQLLSLLSAGRARCVWDPPRPQPGSAGHPPDAQ PGLYYSANEQCRVAFGPKAVACTFAREHLDMCQALSCHTDPLDQSSCSRLLVPLLDGTECGVEKWCSKGRCRSLV ELTPIAAVHGRWSSWGPRSPCSRSCGGGVVTRRRQCNNPRPAFGGRACVGADLQAEMCNTQACEKTQLEFMSQQC ARTDGQPLRSSPGGASFYHWGAAVPHSQGDALCRHMCRAIGESFIMKRGDSFLDGTRCMPSGPREDGTLSLCVSG SCRTFGCDGRMDSQQVWDRCQVCGGDNSTCSPRKGSFTAGRAREYVTFLTVTPNLTSVYIANHRPLFTHLAVRIG GRYVVAGKMSISPNTTYPSLLEDGRVEYRVALTEDRLPRLEEIRIWGPLQEDADIQVYRRYGEEYGNLTRPDITF TYFQPKPRQAWVWAAVRGPCSVSCGAGLRWVNYSCLDQARKELVETVQCQGSQQPPAWPEACVLEPCPPYWAVGD FGPCSASCGGGLRERPVRCVEAQGSLLKTLPPARCRAGAQQPAVALETCNPQPCPARWEVSEPSSCTSAGGAGLA LENETCVPGADGLEAPVTEGPGSVDEKLPAPEPCVGMSCPPGWGHLDATSAGEKAPSPWGSIRTGAQAAHVWTPA AGSCSVSCGRGLMELRFLCMDSALRVPVQEELCGLASKPGSRREVCQAVPCPARWQYKLAACSVSCGRGVVRRIL YCARAHGEDDGEEILLDTQCQGLPRPEPQEACSLEPCPPRWKVMSLGPCSASCGLGTARRSVACVQLDQGQDVEV DEAACAALVRPEASVPCLIADCTYRWHVGTWMECSVSCGDGIQRRRDTCLGPQAQAPVPADFCQHLPKPVTVRGC WAGPCVGQGACGRQHLEPTGTIDMRGPGQADCAVAIGRPLGEVVTLRVLESSLNCSAGDMLLLWGRLTWRKMCRK LLDMTFSSKTNTLVVRQRCGRPGGGVLLRYGSQLAPETFYRECDMQLFGPWGEIVSPSLSPATSNAGGCRLFINV APHARIAIHALATNMGAGTEGANASYILIRDTHSLRTTAFHGQQVLYWESESSQAEMEFSEGFLKAQASLRGQYW TLQSWVPEMQDPQSWKGKEGT SEQ ID NO: 44 Exemplified rSIV Rev protein MPLGPEERRLLRLIWLLYSTNPYPPSGEGTARQRRRARRRWRQQQDQIRVLVERLQEQVYAVDRLADEAQHLAIQ QLPDPPHSA SEQ ID NO: 45 oIC001 DNA forward primer GGGAAAGTGATGTGGTGTACTG SEQ ID NO: 46 oIC002 DNA and RNA reverse primer CAATGGACACAGGGCTGAAA SEQ ID NO: 47 oIC105 RNA forward primer TGCCTTCTCCCTCCTGT SEQ ID NO: 48 CAGGS promoter comprising a chicken β-actin splice donor and a rabbit β-globulin splice acceptor ggagtcgctgcgttgccttcgccccgtgccccgctccgcgccgcctcgcgccgcccgccccggctctgactgacc gcgttactcccacaggtgagcgggcgggacggcccttctcctccgggctgtaattagcgcttggtttaatgacgg ctcgtttcttttctgtggctgcgtgaaagccttaaagggctccgggagggccctttgtgcgggggggagcggctc ggggggtgcgtgcgtgtgtgtgtgcgtggggagcgccgcgtgcggcccgcgctgcccggcggctgtgagcgctgc gggcgcggcgcggggctttgtgcgctccgcgtgtgcgcgaggggagcgcggccgggggcggtgccccgcggtgcg ggggggctgcgaggggaacaaaggctgcgtgcggggtgtgtgcgtgggggggtgagcagggggtgtgggcgcggc ggtcgggctgtaacccccccctgcacccccctccccgagttgctgagcacggcccggcttcgggtgcggggctcc gtgcggggcgtggcgcggggctcgccgtgccgggcggggggtggcggcaggtgggggtgccgggcggggcggggc cgcctcgggccggggagggctcgggggaggggcgcggcggccccggagcgccggcggctgtcgaggcgcggcgag ccgcagccattgccttttatggtaatcgtgcgagagggcgcaagggacttcctttgtcccaaatctgtgcggagc cgaaatctgggaggcgccgccgcaccccctctagcgggcgcggggcgaagcggtgcggcgccggcaggaaggaaa tgggcggggagggccttcgtgcgtcgccgcgccgccgtccccttctccctctccagcctcggggctgtccgcggg gggacggctgccttcgggggggacggggcagggcggggttcggcttctggcgtgtgaccggcagctctagagcct ctgctaaccatgttcatgccttcttctttttcctacag SEQ ID NO: 49 Plasmid as defined in Figure 2H (pDNA3 pMD2.G) GGATCCCCTGAGGGGGCCCCCATGGGCTAGAGGATCCGGCCTCGGCCTCTGCATAAATAAAAAAAATTAGTCAGC CATGAGCTTGGCCCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTA CCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATAT ATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACG TCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGG TAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAA TGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTC ATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTT CCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGT AACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTA GTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGC CTCCCCTCGAAGCTTACATGTGGTACCGAGCTCGGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATG TTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGAC CAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTAT TTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTA AAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTG TAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTATGGTTG GGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCATGTTCATACCTCTTATCTTCCTCCCA CAGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCATCACTTTGGCAAAGCACGTGAGATCTGAATTCAACAG AGATCGATCTGTTTCCTTGACACTATGAAGTGCCTTTTGTACTTAGCCTTTTTATTCATTGGGGTGAATTGCAAG TTCACCATAGTTTTTCCACACAACCAAAAAGGAAACTGGAAAAATGTTCCTTCTAATTACCATTATTGCCCGTCA AGCTCAGATTTAAATTGGCATAATGACTTAATAGGCACAGCCATACAAGTCAAAATGCCCAAGAGTCACAAGGCT ATTCAAGCAGACGGTTGGATGTGTCATGCTTCCAAATGGGTCACTACTTGTGATTTCCGCTGGTATGGACCGAAG TATATAACACAGTCCATCCGATCCTTCACTCCATCTGTAGAACAATGCAAGGAAAGCATTGAACAAACGAAACAA GGAACTTGGCTGAATCCAGGCTTCCCTCCTCAAAGTTGTGGATATGCAACTGTGACGGATGCCGAAGCAGTGATT GTCCAGGTGACTCCTCACCATGTGCTGGTTGATGAATACACAGGAGAATGGGTTGATTCACAGTTCATCAACGGA AAATGCAGCAATTACATATGCCCCACTGTCCATAACTCTACAACCTGGCATTCTGACTATAAGGTCAAAGGGCTA TGTGATTCTAACCTCATTTCCATGGACATCACCTTCTTCTCAGAGGACGGAGAGCTATCATCCCTGGGAAAGGAG GGCACAGGGTTCAGAAGTAACTACTTTGCTTATGAAACTGGAGGCAAGGCCTGCAAAATGCAATACTGCAAGCAT TGGGGAGTCAGACTCCCATCAGGTGTCTGGTTCGAGATGGCTGATAAGGATCTCTTTGCTGCAGCCAGATTCCCT GAATGCCCAGAAGGGTCAAGTATCTCTGCTCCATCTCAGACCTCAGTGGATGTAAGTCTAATTCAGGACGTTGAG AGGATCTTGGATTATTCCCTCTGCCAAGAAACCTGGAGCAAAATCAGAGCGGGTCTTCCAATCTCTCCAGTGGAT CTCAGCTATCTTGCTCCTAAAAACCCAGGAACCGGTCCTGCTTTCACCATAATCAATGGTACCCTAAAATACTTT GAGACCAGATACATCAGAGTCGATATTGCTGCTCCAATCCTCTCAAGAATGGTCGGAATGATCAGTGGAACTACC ACAGAAAGGGAACTGTGGGATGACTGGGCACCATATGAAGACGTGGAAATTGGACCCAATGGAGTTCTGAGGACC AGTTCAGGATATAAGTTTCCTTTATACATGATTGGACATGGTATGTTGGACTCCGATCTTCATCTTAGCTCAAAG GCTCAGGTGTTCGAACATCCTCACATTCAAGACGCTGCTTCGCAACTTCCTGATGATGAGAGTTTATTTTTTGGT GATACTGGGCTATCCAAAAATCCAATCGAGCTTGTAGAAGGTTGGTTCAGTAGTTGGAAAAGCTCTATTGCCTCT TTTTTCTTTATCATAGGGTTAATCATTGGACTATTCTTGGTTCTCCGAGTTGGTATCCATCTTTGCATTAAATTA AAGCACACCAAGAAAAGACAGATTTATACAGACATAGAGATGAACCGACTTGGAAAGTAACTCAAATCCTGCACA ACAGATTCTTCATGTTTGGACCAAATCAACTTGTGATACCATGCTCAAAGAGGCCTCAATTATATTTGAGTTTTT AATTTTTATGGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAATGCCCTGGC CCACAAGTTTCACTAAGCTCGCTTCCTTGCTGTCCAATTTCTATTAAAGGTTCCTTGGTTCCCTAAGTCCAACTA CTAAACTGGGGGATATTATGAAGGGCCTTGAGCATCTGGATTCTGCCTAATAAAAAACATTTATTTTCATTGCAA TGATGTATTTAAATTATTTCTGAATATTTTACTAAAAAGGGAATGTGGGAGGTCAGTGCATTTAAAACATAAAGA AATGAAGAGCTAGTTCAAACCTTGGGAAAATACACTATATCTTAAACTCCATGAAAGAAGGTGAGGCTGCAAACA GCTAATGCACATTGGCAACAGCCCTGATGCCTATGCCTTATTCATCCCTCAGAAAAGGATTCAAGTAGAGGCTTG ATTTGGAGGTTAAAGTTTGGCTATGCTGTATTTTACATTACTTATTGTTTTAGCTGTCCTCATGAATGTCTTTTC ACTACCCATTTGCTTATCCTGCATCTCTCAGCCTTGACTCCACTCAGTTCTCTTGCTTAGAGATACCACCTTTCC CCTGAAGTGTTCCTTCCATGTTTTACGGCGAGATGGTTTCTCCTCGCCTGGCCACTCAGCCTTAGTTGTCTCTGT TGTCTTATAGAGGTCTACTTGAAGAAGGAAAAACAGGGGGCATGGTTTGACTGTCCTGTGAGCCCTTCTTCCCTG CCTCCCCCACTCACAGTGACCCGGAATCCCTCGACATGGCAGTCTAGCACTAGTGCGGCCGCAGATCTGCTTCCT CGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGT TATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAA AGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGA GGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTC CGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCT GTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACC GCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCA CTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCT ACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTT GATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAG GATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTT TGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAA GTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTAT TTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCA GTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGG CCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAA GTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTG GTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGG TTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCAC TGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCT GAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAA CTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCA GTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAA AAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTT TTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGT SEQ ID NO: 50 HIV RRE sequence agtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaataggagctttgttcct tgggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacaggccagacaattatt gtctggtatagtgcagcagcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaactcacagt ctggggcatcaagcagctccaggcaagaatcctggctgtggaaagatacctaaaggatcaacagctcctggggat ttggggttgctctggaaaactcatttgcaccactgctgtgccttggaatgctagt EXAMPLES The invention is now described with reference to the Examples below. These are not limiting on the scope of the invention, and a person skilled in the art would be appreciate that suitable equivalents could be used within the scope of the present invention. Thus, the Examples may be considered component parts of the invention, and the individual aspects described therein may be considered as disclosed independently, or in any combination. Example 1 – Design and production of a β-globin/IgG chimeric intron and insertion into an SIV vector genome plasmid A functional β-globin/IgG chimeric intron comprising an SIV RRE was designed using SnapGene® Software (version 5.3.2, Insightful Science, San Diego, CA, USA; available at www.snapgene.com). To begin, the chimeric intron sequence of the pCI Mammalian Expression Vector (Cat. E1731, Promega, Madison, WI, USA) was analysed using the Basic Local Alignment Search Tool (BLAST, National Centre for Biotechnology Information, Bethesda, MD, USA; https://blast.ncbi.nlm.nih.gov/Blast.cgi). A region homologous to the Hemoglobin Subunit B gene, as well as the splice donor, branch site, polypyrimidine tract, and splice acceptor were mapped onto the chimeric intron. The Rev Response Element (RRE) from r.SIV (Griesenbach et al. (2012) American Journal of Respiratory and Critical Care Medicine 186, 846-856). was moved from upstream of the promoter and inserted in silico within the intron, 5ʹ of the region homologous to Hemoglobin, and 3ʹ to the splice acceptor site. The resulting lentiviral genome plasmid backbone, with the RRE moved from between the partial GAG and cPPT elements to within an intron, was synthesized as a DNA Ultramer with a 5ʹ KpnI and a3ʹ NheI restriction site by IDT (IDT, Newark, NJ, USA). The synthesized Ultramer was double digested with KpnI-HF and NheI restriction enzymes (Cat.# R3142 and R3131, New England BioLabs, Ipswich, MA, USA) following manufacture’s protocols, size separated on a 1% agarose gel, purified with a Monarch® DNA gel extraction kit (Cat.# T1020, New England BioLabs), and ligated with T4 DNA ligase (Cat.# M0202, New England BioLabs) following manufacture’s protocols. Ligation products were heat shock transformed into 5α competent Escherichia coli (Cat. #2987, New England BioLabs), plated on LB plates with 50 μg/mL of kanamycin, and incubated overnight at 37°C. Individual colonies were picked, grown up in 5 mL liquid LB cultures with 50 μg/mL kanamycin. Clones were MiniPrepped using a Monarch® Plasmid MiniPrep Kit (Cat.# T010, New England BioLabs) and screened by PCR, restriction digest, and sanger sequencing. Glycerol stocks were made of a successful clone by combining 500 μL of E. coli in liquid culture with 500 μL of 50% glycerol, and storing immediately at -80°C. Figure 1 shows a schematic of the resulting β-globin/IgG chimeric intron comprising an SIV RRE. Figure 2G shows a plasmid map of the pGM407 SIV vector genome plasmid, which comprises an AAT transgene under the control of a hCEF promoter. The pGM407 plasmid lacks an intron and comprises a SIV RRE. Figure 2A shows a plasmid map of a plasmid derived from pGM407 which has had the SIV RRE deleted and the β-globin/IgG chimeric intron comprising an SIV RRE of Figure 1 inserted between with hCEF promoter and the AAT transgene, as described above. Example 2 – Transfection of HEK293T cells with a plasmid comprising a β-globin/IgG chimeric intron with an inserted SIV RRE increases AAT transgene expression HEK293T cells were transfected with a plasmid lacking the endogenous SIV RRE and comprising (i) an AAT transgene under the control of an hCEF promoter and (ii) a β-globin/IgG chimeric intron with an inserted SIV RRE as produced in Example 1 (pGM991). As shown in Figure 3, transfection of HEK293T cells with pGM991 significantly increased AAT expression compared with transfection of HEK293T cells with a corresponding plasmid comprising the endogenous SIV RRE and lacking the β-globin/IgG chimeric intron with an inserted SIV RRE (pGM407). In particular, an increase in AAT expression of 10.6-fold was observed on transfection with the plasmid lacking the endogenous SIV RRE and comprising the β-globin/IgG chimeric intron with an inserted SIV RRE compared with transfection with a corresponding plasmid comprising the endogenous SIV RRE and lacking said intron/RRE. PCR of DNA extracted from the transfected HEK293T cells was carried out using primers that bind to either side of the intron:

As shown in Figure 4A, the intron in pGM991 was present and a large 1127 bp product was detected. In contrast, pGM407 produced a smaller product of 371 bp. RNA was extracted from the HEK293T cells transfected with either pGM991 or pGM407. RT- PCR was carried out on the extracted RNA using primers which bind to either side of the intron:

As shown in Figure 4B, the intron was spliced successfully by the transfected HEK293T cells, such that both pGM991 and pGM407 produced the same 277 bp RNA transcript. Example 3 – SIV vector comprising a β-globin/IgG chimeric intron with an inserted SIV RRE is correctly packaged and spliced HEK293T cells were transduced with (a) a VSV-G pseudotyped SIV vector (SIV.VSV-G) lacking the endogenous SIV RRE and comprising (i) an AAT transgene under the control of an hCEF promoter and (ii) a β-globin/IgG chimeric intron with an inserted SIV RRE as produced in Example 1 (vGM291); or (b) a SIV.VSV-G vector comprising the endogenous SIV RRE and lacking this chimeric intron-RRE (vGM290). Non-transduced cells were used as controls. PCR of DNA extracted from the transduced HEK293T was carried out using primers which bind to either side of the intron:

As shown in Figure 5, vGM291 was properly packaged to produce SIV.VSV-G particles, with an 1127 bp product being detected. In contrast, vGM290 produced a smaller product of 371 bp. RNA was extracted from the transduced HEK293T cells transduced with either vGM291; or vGM290. RT-PCR was carried out on the extracted RNA using primers which bind to either side of the intron:

As shown in Figure 6, the intron was spliced successfully by the transduced HEK293T cells, such that both vGM290 and vGM291 produced the same 277 bp RNA transcript. The effect of inclusion of the β-globin/IgG chimeric intron with an inserted SIV RRE on AAT transgene expression was also investigated. As shown in Figure 7, transduction of HEK293T cells with this SIV.VSV-G vector significantly increased AAT expression compared with transduction of HEK293T cells with a corresponding SIV.VSV-G vector comprising the endogenous SIV RRE and lacking the β- globin/IgG chimeric intron with an inserted SIV RRE (vGM290). In particular, an increase in AAT expression of 686-fold was observed on transduction with vGM291 compared with transduction with vGM290. Example 4 – SIV vector comprising a β-globin/IgG chimeric intron with an inserted SIV RRE increases AAT transgene transcription Furthermore, the effect of the intron on transgene transcription was investigated. Reverse transcription droplet digital PCR (RT-ddPCR) was carried out on RNA extracted from HEK293T cells transfected with plasmids as described in Example 2. Amount of mRNA was quantified by the number of WPRE copies using RT-ddPCR, standardised to the housekeeping gene β-2-microglobulin (B2M). As shown in Figure 8A, use of the pGM991 plasmid comprising the β-globin/IgG chimeric intron with an inserted SIV RRE significantly increased the amount of mRNA produced by the HEK293T cells compared with use of the pGM407 plasmid which lacks the β-globin/IgG chimeric intron with an inserted SIV RRE. Furthermore, the amount of mRNA produced per plasmid copy was also calculated, for which plasmid copy number was quantified by ddPCR from DNA extracted from transfected HEK293T cells. As shown in Figure 8B, use of the pGM991 plasmid significantly increased the number of mRNA copies per plasmid copy compared to the pGM407 plasmid. Example 5 – SIV.FN/H vector comprising a β-globin/IgG chimeric intron with an inserted SIV RRE increases AAT transgene transcription The experiment in Example 3 was repeated with an F/HN pseudotyped SIV vector, rather than a VSV-G pseudotyped SIV vector. As shown in Figure 9, transduction of HEK293T cells with a SIV.F/HN vector containing the β-globin/IgG chimeric intron (vGM295) significantly increased AAT expression compared to HEK293T cells transduced with a corresponding SIV.F/HN vector comprising the endogenous SIV RRE and lacking the β-globin/IgG chimeric intron (vGM294). In particular, an increase in AAT expression of 501-fold was observed on transduction with vGM295 compared to transduction with vGM294. Thus, the use of the β-globin/IgG chimeric intron with an inserted SIV RRE facilitates the maturation of a stable mRNA molecule, resulting in increased mRNA transcript levels, and consequently increased transgene expression, and this effect is observed in multiple different pseudotyped SIV vectors. Whilst this is exemplified with AAT, it is expected that similar advantages will be achieved using other transgenes.

Claims

CLAIMS 1. A retroviral vector comprising an intron; wherein: (a) the endogenous Rev response element (RRE) of the retroviral genome is deleted; and (b) a retroviral RRE is inserted into the intron within 100 bp 5’ of the splice acceptor’s branch site of the intron.

2. A retroviral vector according to claim 1, wherein the retroviral RRE is inserted within 20 bp 5’ of the splice acceptor’s branch site of the intron.

3. A retroviral vector according to claim 1 or 2, wherein: (a) the intron is a chimeric intron, optionally selected from a β-globin/IgG chimeric intron or a chimeric intron from the CAGGS promoter; or (b) the intron is a viral intron, optionally selected from SV40 intron, CMV Intron A and adenovirus tripartite leader sequence intron.

4. A retroviral vector comprising a chimeric intron; wherein: (a) the endogenous RRE of the retroviral genome is deleted; and (b) a retroviral RRE is inserted into the chimeric intron.

5. A retroviral vector according to any one of the preceding claims, wherein the retroviral RRE inserted into the intron is the endogenous RRE of the retroviral genome.

6. A retroviral vector according to any one of the preceding claims, wherein the RRE is a Simian immunodeficiency virus (SIV) RRE.

7. A retroviral vector according to claim 6, wherein the RRE comprises or consists of a nucleic acid sequence having at least 90% identity to SEQ ID NO: 1.

8. A retroviral vector according to any one of the preceding claims, wherein the intron is less than 1,000bp in length, preferably less than 800 bp in length.

9. A retroviral vector according to any one of claims 3 to 8, wherein the chimeric intron is a β- globin/IgG chimeric intron or a chimeric intron from the CAGGS promoter.

10. A retroviral vector according to claim 9, wherein the chimeric intron is a β-globin/IgG chimeric intron and the RRE is inserted between (i) a splice donor site comprising or consisting of a nucleic acid sequence of TGAGTTTAAGGTAAGT (SEQ ID NO: 2); and (ii) a splice acceptor site comprising or consisting of a nucleic acid sequence of CTCTCCACAG (SEQ ID NO: 3).

11. A retroviral vector according to claim 9 or 10, wherein the β-globin/IgG chimeric intron comprises or consists of a nucleic acid sequence having at least 90% identity to SEQ ID NO: 4.

12. A retroviral vector according to any one of the preceding claims, wherein the intron is a β- globin/IgG chimeric intron and the RRE is an SIV RRE, and optionally wherein the chimeric intron comprising the RRE comprises or consists of a nucleic acid sequence having at least 90% identity to SEQ ID NO: 5.

13. A retroviral vector according to any one of the preceding claims, wherein the intron is between a promoter and a transgene operably linked to said promoter, wherein optionally the promoter is selected the group consisting of a cytomegalovirus (CMV) promoter, elongation factor 1a (EF1a) promoter, and a hybrid human CMV enhancer/EF1a (hCEF) promoter, preferably a hCEF promoter.

14. A retroviral vector according to any one of claims 13, wherein the transgene encodes a therapeutic protein, wherein optionally said therapeutic protein is selected from: (a) a secreted therapeutic protein, optionally Alpha-1 Antitrypsin (AAT), Factor VIII, Surfactant Protein B (SFTPB), ADAMTS13, Factor VII, Factor IX, Factor X, Factor XI, von Willebrand Factor, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), decorin, Surfactant Protein C (SP-C), an anti-inflammatory protein and a monoclonal antibody against an infectious agent; or (b) CFTR, ABCA3, DNAH5, DNAH11, DNAI1, DNAI2, CSF2RA, CSF2RB and TRIM-72.

15. A retroviral vector according to any one of the preceding claims, wherein the retroviral vector is a lentiviral vector.

16. A retroviral vector according to claim 15, wherein the lentiviral vector is selected from the group consisting of a Human immunodeficiency virus (HIV) vector, a Simian immunodeficiency virus (SIV) vector, a Feline immunodeficiency virus (FIV) vector, an Equine infectious anaemia virus (EIAV) vector, and a Visna/maedi virus vector.

17. A retroviral vector according to any one of the preceding claims, which is pseudotyped with haemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus or G glycoprotein from Vesicular Stomatitis Virus (G-VSV).

18. A retroviral vector according to any one of claims 13 to 17, which increases transgene expression by at least about 2-fold, preferably at least about 5-fold, more preferably at least about 10-fold compared with a corresponding vector which lacks a intron into which a retroviral RRE has been inserted.

19. A nucleic acid comprising or consisting of a intron into which a retroviral RRE has been inserted, wherein optionally (i) the intron; and/or (ii) the RRE are as defined in any one of claims 1 to 12.

20. A plasmid comprising a nucleic acid according to claim 19.

21. A retroviral vector according to any one of claims 1 to 18, a nucleic acid according to claim 19 or a plasmid according to claim 20, which is codon-optimised.

22. A composition comprising a retroviral vector according to any one of claims 1 to 18 or 21, a nucleic acid according to claim 19 or 21 or a plasmid according to claim 20 or 21, and a pharmaceutically-acceptable carrier.

23. A host cell comprising a retroviral vector according to any one of claims 1 to 18 or 21, a nucleic acid according to claim 19 or 21 or a plasmid according to claim 20 or 21.

24. A retroviral vector according to any one of claims 1 to 18 or 21, a nucleic acid according to claim 19 or 21, a plasmid according to claim 20 or 21, or a composition according to claim 22 for use in a method of treatment.

25. A method of producing a retroviral vector, said method comprising the following steps: (a) growing cells in suspension; (b) transfecting the cells with one or more plasmids; (c) adding a nuclease; (d) harvesting the lentivirus; (e) adding trypsin; and (f) purification; wherein the one or more plasmids comprises a vector genome plasmid which comprises a nucleic acid as defined in claim 19, and optionally (i) a promoter as defined in claim 13 and/or (ii) a transgene as defined in claim 14.

26. A method of differentiating between a retroviral vector and a transgene expressed by said retroviral vector, said method comprising the steps of: (a1) transducing cells with a retroviral vector as defined in any one of claims 1 to 18; (b1) culturing the cells to allow transgene expression by the retrovirus; and (c1) quantifying RNA within the cells; or (a2) quantifying RNA within cells of a sample obtained from a patient who has undergone treatment with a retroviral vector according to any one of claims 1 to 18 or 21, a nucleic acid according to claim 19 or 21, a plasmid according to claim 20 or 21, or a composition according to claim 23 ; wherein: (i) the amount of RNA comprising the chimeric intron into which a retroviral RRE has been inserted corresponds to the copy number of the retroviral vector; and (ii) the amount of RNA lacking the chimeric intron into which a retroviral RRE has been inserted corresponds to the amount of transgene mRNA; wherein optionally RNA is quantified by a PCR-based or in situ hybridisation-based assay.