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KR20210056936A - 아데노바이러스 벡터를 포함하는 폐암 진단용 조성물 및 이의 용도 - Google Patents

아데노바이러스 벡터를 포함하는 폐암 진단용 조성물 및 이의 용도 Download PDF

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KR20210056936A
KR20210056936A KR1020200150172A KR20200150172A KR20210056936A KR 20210056936 A KR20210056936 A KR 20210056936A KR 1020200150172 A KR1020200150172 A KR 1020200150172A KR 20200150172 A KR20200150172 A KR 20200150172A KR 20210056936 A KR20210056936 A KR 20210056936A
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최진우
이수민
박종만
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주식회사 코어파마
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Abstract

본 발명은 상피세포에 감염능이 있는 아데노바이러스를 이용한 것으로, SFTP (Surfactant Protein) 유전자의 프로모터; 표지 유전자로는 GFP (Green Fluorescent Prtein)를 이용하였고, 또한 혈중에 대다수로 존재하는 혈구 세포에서는 발현하지 않고, 폐암세포에서만 발현할 수 있도록 miR-142를 표적하는 서열을 배열하여, 폐암 세포에 높은 특이적 발현성을 가진다. 따라서, 본 발명의 아데노바이러스는 혈중에 돌아다니는 폐암 세포에 특이적으로 형광을 발광하게 하여, 폐암 조기진단 및 치료 유용하게 활용될 수 있다.

Description

아데노바이러스 벡터를 포함하는 폐암 진단용 조성물 및 이의 용도 {Lung cancer diagnostic composition comprising an adenovirus vector and use thereof}
본 발명은 아데노바이러스 벡터 (Adenovirus vector)를 포함하는 폐암 (Lung cancer) 진단용 조성물 및 이의 용도에 관한 것이다.
우리나라 암 (악성신생물) 사망자 수는 2002년 우리나라 총 사망자 246,515명 (조사망률 인구 10만 명당 512명) 가운데 25.5% (남성 사망자의 29.6%, 여성 사망자의 20.5%)인 62,887명으로 암으로 인한 사망 (사망률 인구 10만명당 130.7명)이 사망원인 1위를 차지하고 있다. 암 사망순위는 폐암, 위암, 폐암, 대장암 및 췌장암 순으로, 이들 5대 암에 의한 사망이 전체 암 사망의 약 70%를 차지하고 있다. 또한 남성에 있어 주요 암 사망원인은 폐암, 위암, 폐암 및 대장암 등으로 이들 4대 암에 의한 사망자 수 (28,147명)가 전체 남성 암 사망자 수 (40,177명)의 70%를 차지하고 있으며, 여성에 있어 주요 암 사망원인은 위암, 폐암, 폐암, 대장암 및 췌장암 등으로 이들 5대 암에 의한 사망자 수 (13,630명)가 전체 여성 암 사망자 수 (22,710명)의 60%를 차지하고 있다.
이러한 암의 종류는 현재까지 밝혀진 것만 해도 수십 종에 이르며, 주로 발병 조직의 위치에 따라 구분된다. 암은 성장속도가 매우 빠르고, 주변 조직에 침윤하면서 전이 (metastasis)가 일어나 생명을 위협하게 된다. 이러한 암의 종류로는 뇌척수종양, 두경부암, 폐암, 유방암, 흉선종, 식도암, 취암, 대장암, 폐암, 췌장암, 담도암, 신장암, 방광암, 폐암, 고환암, 생식세포종, 난소암, 자궁 경부암, 자궁 내막암, 림프종, 급성 백혈병, 만성 백혈병, 다발성 골수종, 육종, 악성 흑색종 및 피부암등이 있다. 또한 이러한 암의 발병 기전 또는 형태에 따라 다른 분류 체계에 의해 구분되기도 한다.
특히, 폐암은 전 세계적으로 암으로 인한 사망 원인의 30%를 차지할 정도로 암으로 인한 사망의 주요 원인 중의 하나이다. 전체 폐암의 75% 이상이 비소세포폐암 (non-small cell lung cancer, NSCLC)이며, 평균 5년 생존율이 15%에 해당한다. 폐암의 높은 사망률은 일부 절제 불가능한 종양을 가진 환자의 높은 비율과 관련되어 있으며, 또한 절제 가능한 병기의 비소세포폐암 환자 중에서도 외과적으로 절제한 환자의 상당수가 암의 재발로 사망하고 있다. 현재 폐암의 경우 병리학적 병기로 진단하고 있지만, 같은 병기에서도 재발과 생존의 상당한 차이를 보이고 있기 때문에, 폐암의 진단과 치료의 방법이 발달했음에도 불구하고 여전히 생존율이 좋지 않다. 이러한 결과는 조기 진단방법과 알맞은 선별검사의 부족으로 인한 낮은 치료율에 기인하며, 따라서 폐암 사망률을 낮추기 위하여 효과적인 조기 진단 및 맞춤진단방법의 개발이 필요한 실정이다.
폐 계면활성제는 폐포의 표면을 덮고 있는 공기-액체의 경계면에 존재하여 공기-액체 사이에 생기는 강한 표면장력 (surface tension)을 저하시켜 주는 작용을 하여 폐가 collapse되는 것을 방지해 주는 물질로서, 제 Ⅱ형 폐포 (type Ⅱ pneumocyte)로 부터 생산, 분비되어 폐포에 필름 (film)을 형성하여 작용한다. 폐 계면활성제는 제 Ⅱ형 폐포내에 존재하는 것과 폐포의 표면에 존재하는 것 두가지가 있는데, 폐포 내에 존재하는 폐 계면활성제의 추출은 어려운 점이 많아 폐포의 표면에 존재하는 것을 생리식염수로 폐 세정 (lavage)하여 추출하였을 때 생체내의 물질과 가장 가까운 폐 계면활성제를 획득할 수 있으므로 이 방법으로 획득한 폐 계면활성제로 생화학적 구성 및 표면활성의 물리적 특성이 규명되기 시작하였다. 10% 정도의 구성비를 차지하고 있는 단백질 중에는 오염된 혈청 단백질이 8% 정도이고 이것을 제외한 순수한 폐 계면활성제 자체의 단백질은 1% 정도이고 이를 pulmonary surfactant associated protein (PSP, 또는 apoprotein)이라고 불리어 오다가 1987년부터 surfactant protein (SP)이라고 통일되어 불리어진다.
일반적으로 분자량에 따라 3가지 군으로 나뉘는데 분자량이 28,000~35,000 Da인 SP-A, 7,000~8,000 Da인 SP-B와 5,000 Da인 SP-C가 있다. SP-A는 수용성이고 (hydrophilic), SP-B 및 SP-C는 유기용매 가용의 소수성 (hydrophobic)이다. 최근에는 4번째 단백질인 SP-D의 존재 및 역할에 관한 보고가 있다. SP-A는 폐 계면활성제의 표면활성의 물리적 작용을 돕는 것 보다는 폐 계면활성제의 제 Ⅱ형 폐포내의 존재형태인 tubular myelin의 지질 대사와 폐 계면활성제의 생산, 분비, 흡수 및 폐포 방어 기전과 면역 기전에 관계한다고 알려져 있다. 임상에서 폐 계면활성제의 보충요법을 실시할 때 사용하는 calf lung surfactant extract (CLSE)와 Surfactenⓡ은 SP-A를 함유하고 있지 않은 제제인데 이 두가지 제제가 in vitro 표면 활성도가 높고 임상적 개선도가 높아 SP-A는 폐 계면활성능에 필요치 않은 것으로 보고되고 있다. 소수성 단백질 SP-B와 SP-C는 폐 계면활성제의 폐포 내에서의 film 형성 및 폐포 표면에서의 폐 계면활성제의 흡착을 항진시켜폐 계면활성제의 표면활성 작용에 필수적인 것으로 알려져 있다. 폐 계면활성제 단백질 A1, A2, B, C 및 D는 각각 SFTPA1, SFTPA2, SFTPB, SFTPC 및 SFTPD 유전자에 의해 코딩되는 단백질이다.
한편, 기존의 순환종양세포를 진단하는 방식은 어떤 암종에서 유래된 종양세포인지를 판별하기 어려워서 실제 진단의 효용성이 낮았다. 따라서 바이러스를 이용하여 혈중에 돌아다니는 암세포를 잡아내었을 경우에 환자에게 어떤 암인지를 알려줄수 있다면 조기 진단 및 치료에 도움이 될것으로 생각된다.
이에 본 발명자는, SFTPA2 (Surfactant Protein A2) 프로모터 또는 SFTPC (Surfactant Protein C) 프로모터를 아데노바이러스의 E1a 및 E1b와 작동가능하게 연결하고, 아데노바이러스 E3 부위에 GFP (Green Fluorescent Prtein)를 도입하되, 상기 GFP 말단에 암세포에서만 발현 할 수 있도록 miR-142를 표적하는 서열을 도입하여, 폐암 세포에 높은 특이적 발현성을 확보함으로써, 본 발명을 완성하였다. 이는 순환종양세포를 진단하는 방식에 활용될 수 있으며, 이를 통해 어떠한 암종에서 유래된 종양세포인지를 판별할 수 있을 것으로 보인다.
본 발명은 상기와 같은 문제를 해결하기 위해, 폐암 세포에 높은 특이적 발현성을 가지는 아데노바이러스 벡터를 발명하여, 본 발명의 아데노바이러스를 이용할시 혈중에 돌아다니는 폐암 세포를 잡아내어 형광을 띄게 만드는 방법으로 조기진단 및 치료에 도움을 주는 것을 목적으로 한다.
상기한 과제를 해결하기 위하여, 본 발명은 SFTP(Surfactant Protein) 유전자의 프로모터, 표지 유전자 및 miR 142를 표적하는 염기서열을 포함하는 바이러스 벡터를 제공한다.
본 발명의 일 실시예에 있어서, “SFTP 유전자 프로모터”는 SFTPA2 (Surfactant Protein A2) 유전자 프로모터 또는 SFTPC (Surfactant Protein C) 유전자 프로모터인 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “SFTPA2 (Surfactant Protein A2) 유전자 프로모터”는 서열번호 5로 표시되고, “SFTPC (Surfactant Protein C) 유전자 프로모터”는 서열번호 6으로 표시되는 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “표지”는 루시퍼라아제 (luciferase), 증강 녹색 형광 단백질 (enhanced green fluorescent protein, EGFP), 녹색 형광 단백질 (green fluorescent protein, GFP), 노랑 형광 단백질 (Yellow Fluorescent Protein, YFP), 적색 형광 단백질 (Red Fluorescent Protein, RFP)및 청색 형광 단백 (cyan fluorescent protein, CFP)으로 이루어진 군에서 선택된 1종 이상인 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “miR-142를 표적하는 서열”은 miR-142-3p 표적서열인 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “miR-142-3p 표적서열”은 서열번호 7로 표시되는 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “miR-142를 표적하는 서열”은 표지 유전자의 5’UTR 부위에 위치하는 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “벡터”는 SFTP에 작동가능하게 연결된 인핸서를 포함하는 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “벡터의 구조”는 SFTP 유전자 프로모터, E1a, E1b, L1, L2, E2, L3, L4, E3 및 E4가 순차적으로 배열된 것을 특징으로 할 수 있다.
본 발명의 일시예에 따르면, 더욱 구체적인 “벡터의 구조”는 SFTP 유전자 프로모터, E1a, E1b, L1, L2, E2, L3, L4, E3, 표지 프로모터, 상기 표지 프로모터에 작동가능하게 연결된 표지 유전자, 상기 표지 유전자의 5’UTR 부위에 도입된 miR-142를 표적하는 서열 및 E4가 순차적으로 배열된 것일 수 있다.
본 발명의 일 실시예에 있어서, “벡터”는 폐암세포에서 특이적으로 표지 유전자를 발현하는 것을 특징으로 할 수 있다.
본 발명의 일 실시예에 있어서, “벡터”는 서열번호 8 또는 서열번호 9로 표시되는 것을 특징으로 할 수 있다.
또한, 본 발명은 본 발명의 바이러스 벡터를 포함하는 폐암 진단용 키트를 제공한다.
또한, 본 발명은 본 발명의 바이러스 벡터를 포함하는 폐암 진단용 조성물을 제공한다.
또한, 본 발명은 폐암세포를 검출하는 방법을 제공한다.
또한, 본 발명은 순환폐암세포의 분리 및 배양 방법을 제공한다.
마지막으로, 본 발명은 폐암의 진단에 필요한 정보를 제공하는 방법을 제공한다.
본 발명의 상피세포에 감염능이 있는 아데노바이러스는 혈중에 돌아다니는 폐암 세포에서 특이적으로 형광을 발현하므로, 이를 이용하여 폐암의 조기진단 및 치료에 이용할 수 있다.
도 1은 본 발명의 코스미드 벡터 제작의 개략도이다.
도 2는 본 발명의 아데노바이러스의 구조를 모식화한 도이다.
도 3은 재조합 벡터의 암세포 특이적 발현 효과를 확인한 도이다.
이하, 첨부된 도면을 참조하여 본 발명의 구현예로 본 발명을 상세히 설명하기로 한다. 다만, 하기 구현 예는 본 발명에 대한 예시로 제시되는 것으로, 당업자에게 주지 저명한 기술 또는 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우에는 그 상세한 설명을 생략할 수 있고, 이에 의해 본 발명이 제한되지는 않는다. 본 발명은 후술하는 특허 청구범위의 기재 및 그로부터 해석되는 균등 범주 내에서 다양한 변형 및 응용이 가능하다.
또한, 본 명세서에서 사용되는 용어 (terminology)들은 본 발명의 바람직한 실시 예를 적절히 표현하기 위해 사용된 용어들로서, 이는 사용자, 운용자의 의도 또는 본 발명이 속하는 분야의 관례 등에 따라 달라질 수 있다. 따라서 본 용어들에 대한 정의는 본 명세서 전반에 걸친 내용을 토대로 내려져야 할 것이다. 명세서 전체에서, 어떤 부분이 어떤 구성요소를 “포함”한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.
본 명세서 전체에 걸쳐, 특정 물질의 농도를 나타내기 위하여 사용되는 '%'는 별도의 언급이 없는 경우, 고체/고체는 (w/w) %, 고체/액체는 (w/v) %, 그리고 액체/액체는 (v/v) %이다.
일 측면에서, 본 발명은 SFTP(Surfactant Protein) 유전자의 프로모터, 표지 유전자 및 miR 142를 표적하는 염기서열을 포함하는 바이러스 벡터를 제공한다.
본 발명에서 용어, "바이러스 벡터"는 원하는 세포, 조직 및/또는 기관으로 치료 유전자 또는 유전물질을 전달할 수 있는 바이러스 벡터이다.
일 구현예에서, SFTP 유전자 프로모터는 SFTPA2(Surfactant Protein A2) 유전자 프로모터 또는 SFTPC(Surfactant Protein C) 유전자 프로모터일 수 있으며, SFTPA2 유전자의 프로모터는 서열번호 5로 표시되는 염기서열을 포함할 수 있고, SFTPAC 유전자의 프로모터는 서열번호 6으로 표시되는 염기서열을 포함할 수 있다.
일 구현예에서, 상기 벡터는 SFTP에 작동가능하게 연결된 인핸서를 포함할 수 있다.
일 구현예에서, 상기 바이러스 벡터는 아데노바이러스 (Adenovirus), 아데노 조력 바이러스 (Adeno-associated virus), 렌티바이러스, 레트로바이러스, HIV (Human immunodeficiency virus), MLV (Murine leukemia virus), ASLV (Avian sarcoma/leukosis), SNV (Spleen necrosis virus), RSV (Rous sarcoma virus), MMTV (Mouse mammary tumor virus) 및 헤르페스 심플렉스 바이러스 (Herpes simplex virus)로 이루어진 군 중에서 선택된 1종 이상을 포함할 수 있으나, 아데노바이러스 벡터인 것이 가장 바람직하다.
일 구현예에서, 상기 바이러스 벡터는 아데노바이러스 벡터일 수 있으며, 이의 E3 영역 내에 표지 유전자 및 miR 142를 표적하는 염기서열을 포함할 수 있고, E1 영역 내 핵산 서열의 결실을 포함할 수 있으며, IRES 서열이 E1 영역 내 삽입됨으로써 E1a 및 E1b로 나뉠 수 있다.
본 발명에서 사용되는 용어 "결실"은 하나 이상의 아미노산 또는 뉴클레오티드의 삭제를 말하며, 본 발명의 일 실시예에서는 아데노바이러스의 E1 유전자 (영역)를 결실시켰다.
일 구현예에서, SFTP 유전자의 프로모터가 아데노바이러스의 내재적 유전자와 작동 가능하게 연결될 수 있으며, 아데노바이러스의 내재적 유전자는 E1a, E1b, L1, L2, E2, L3, L4, E3 또는 E4를 포함할 수 있다.
일 구현예에서, 본 발명의 벡터의 구조는 SFTP 유전자 프로모터, E1a, E1b, L1, L2, E2, L3, L4, E3 및 E4가 순차적으로 배열될 수 있으며, SFTP 유전자 프로모터, E1a, E1b, L1, L2, E2, L3, L4, E3, 표지 프로모터, 상기 표지 프로모터에 작동가능하게 연결된 표지 유전자, 상기 표지 유전자의 5’UTR 부위에 도입된 miR-142를 표적하는 서열 및 E4가 순차적으로 배열될 수 있다.
본 발명에서 용어, "작동가능하게 연결된 (operably linked)"은 일반적 기능을 수행하도록 핵산 발현조절 서열과 목적하는 단백질 또는 RNA를 코딩하는 핵산 서열이 기능적으로 연결 (functional linkage)되어 있는 것을 말한다. 예를 들어 프로모터와 단백질 또는 RNA를 코딩하는 핵산 서열이 작동가능하게 연결되어 코딩하는 핵산서열의 발현에 영향을 미칠 수 있다. 재조합 벡터와의 작동적 연결은 당해 기술분야에서 잘 알려진 유전자 재조합 기술을 이용하여 제조할 수 있으며, 부위-특이적 DNA 절단 및 연결은 당해 기술 분야에서 일반적으로 알려진 효소 등을 사용한다.
일 구현예에서, miR 142를 표적하는 염기서열은 miR-142-3p 표적서열일 수 있으며, 서열번호 7로 표시되는 염기서열을 포함할 수 있다.
본 발명에서 용어, "마이크로 RNA (miRNA)"는 MicroRNA (약칭 miR)는 약 22개 nucleotide로 이루어진 noncoding RNA로 유전자 발현을 조절하는 역할을 한다. 상기 microRNA는 유전자의 전사 후 post-transcription 단계에서 작용하며, 포유류의 경우 유전자의 60% 정도가 microRNA에 의해 발현이 조절되는 것으로 알려져 있다. MicroRNA는 다양한 생체 내 프로세스에서 중요한 역할을 하며 암, 심장 질환, 신경 관련 질병에 연관이 있는 것으로 밝혀져 있다. miRNA 는 1 개 사슬 RNA 이지만, premature 의 2 개 사슬 RNA 중 목적 유전자의 발현을 억제할 수 있는 한, 어느 측의 1 개 사슬 RNA 의 표적 배열을 사용할 수 있다. 예컨대, miR-142에는 miR-142-3p 및 miR-142-5p가 존재하며, 본 발명에 있어서는 어느 표적 배열을 사용해도 되며, miR-142에는 miR-142-3p 및 miR-142-5p의 양자가 포함되고, 바람직하게는 miR-142-3p일 수 있다.
본 발명에서 용어, "miR-142-3p"는 이의 유전자가 B 세포 백혈병(aggressive B cell leukemia)에 있어서 전좌가 일어나는 부위에 존재하고 있으며, 조혈 조직 (골수, 비장, 흉선 등)에서 발현하고 있다고 알려져 있다. 또한, miR-142-3p 는 마우스 태아 간장 (태아의 조혈 조직)에 있어서 발현이 확인되어, 조혈계의 분화에 관여하고 있는 것으로 알려져 있다.
일 구현예에서, 표지는 발색효소, 방사성 동위원소, 크로모포어(chromopore), 발광물질, 형광물질, 프로브 또는 택(tag)을 포함할 수 있으며, 형광물질은 Cy(cyanine) 계열, 로다민(Rhodamine) 계열, 알렉사(Alexa) 계열, BODIPY 계열 또는 ROX 계열의 형광물질일 수 있으며, 루시퍼라아제 (luciferase), 증강 녹색 형광 단백질 (enhanced green fluorescent protein, EGFP), 녹색 형광 단백질 (green fluorescent protein, GFP), 노랑 형광 단백질 (Yellow Fluorescent Protein, YFP), 적색 형광 단백질 (Red Fluorescent Protein, RFP) 또는 청색 형광 단백 (cyan fluorescent protein, CFP)일 수 있다.
일 구현예에서, 상기 표지는 형광 물질, 프로브 또는 택(tag)일 수 있다. 상기 표지 물질은 본 기술 분야에서 알려진 어떠한 방법에 의해 검출될 수 있는 어떠한 화학적 모이어티라도 될 수 있다. 검출가능한 표지의 예는 분광(spectroscopy), 광화학(photochemistry)에 의해 또는 생화학적(biochemical), 면역학적(immunochemical) 또는 화학적 수단에 의해 검출될 수 있는 어떠한 모이어티라도 포함될 수 있다. 핵산 프로브에 표지하는 적절한 방법은 상기 표지의 종류 및 상기 표지의 위치 및 프로브에 따라 선택될 수 있다. 표지의 예는 효소, 효소 기질, 동위원소물질, 형광 다이, 발색단(chromophores), 화학발광 표지(chemiluminescent label), 전기화학적 발광 표지(electrochemical luminescent label), 특이적인 결합 파트너를 갖는 리간드, 및 서로 반응하여 검출 신호의 강도를 증가, 변형 또는 감소시킬 수 있는 기타 표지를 포함할 수 있다.
본 발명에서 용어, "프로브"란 mRNA와 특이적 결합을 이룰 수 있는 짧게는 수 염기 내지 길게는 수백 염기에 해당하는 RNA 또는 DNA 등의 핵산 단편을 의미하며 라벨링 되어 있어서 특정 mRNA의 존재 유무를 확인할 수 있다. 프로브는 올리고 뉴클레오타이드(oligonucleotide) 프로브, 단쇄 DNA(single stranded DNA) 프로브, 이중쇄 DNA(double stranded DNA) 프로브, RNA 프로브 등의 형태로 제작될 수 있다. 본 발명에서는 상기 AFP, HMMR, NXPH4, PITX1, THBS4 및/또는 UBE2T 유전자와 상보적인 프로브를 이용하여 혼성화를 실시하여, 혼성화 여부를 통해 상기 유전자 발현 정도를 진단할 수 있다. 적당한 프로브의 선택 및 혼성화 조건은 통상의 기술분야에 공지된 것을 기초로 변형할 수 있으므로 본 발명에서는 이에 대해 특별히 한정하지 않는다.
본 발명에서, 프로브를 cDNA 분자와 혼성화시키는 적합한 조건은 최적화 절차에 의하여 일련의 과정으로 결정될 수 있다. 이런 절차는 연구실에서 사용을 위한 프로토콜을 수립하기 위하여 당업자에 의하여 일련의 과정으로 실시된다. 예를 들어, 온도, 성분의 농도, 혼성화 및 세척 시간, 완충액 성분 및 이들의 pH 및 이온세기 등의 조건은 프로브의 길이 및 GC 양 및 타깃 뉴클레오타이드 서열 등의 다양한 인자에 의존한다. 혼성화를 위한 상세한 조건은 Joseph Sambrook, et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.(2001); 및 M.L.M. Anderson, NucleicAcidHybridization, Springer-Verlag New York Inc. N.Y.(1999)에서 확인할 수 있다. 예를 들어, 상기 엄격조건 중에서 고 엄격조건은 0.5 M NaHPO4, 7% SDS(sodium dodecyl sulfate),1mM EDTA에서 65℃ 조건으로 혼성화하고, 0.1 x SSC(standard saline citrate)/0.1% SDS에서 68℃ 조건으로 세척하는 것을 의미한다. 또는, 고 엄격조건은 6 x SSC/0.05% 소듐 파이로포스페이트에서 48℃ 조건으로 세척하는 것을 의미한다. 저 엄격조건은 예를 들어, 0.2 x SSC/0.1% SDS에서 42℃ 조건으로 세척하는 것을 의미한다.
일 구현예에서, 상기 바이러스 벡터는 E1a 및 E1b와 작동가능하게 연결된 SFTP 유전자 프로모터; 표지 유전자 및 miR-142를 표적하는 서열을 순차적으로 포함하는 아데노바이러스 재조합 벡터일 수 있다.
본 발명에서 용어, "재조합 벡터"란 적당한 숙주세포에서 목적 단백질 또는 목적 RNA을 발현할 수 있는 벡터로서, 유전자 삽입물이 발현되도록 작동가능하게 연결된 필수적인 조절 요소를 포함하는 유전자 작제물을 말한다.
일 구현예에서, 본 발명의 바이러스 벡터는 폐암세포에서 특이적으로 표지 유전자를 발현할 수 있으며, 서열번호 8 또는 9로 표시되는 염기서열을 포함할 수 있다.
본 발명의 벡터 시스템은 당업계에 공지된 다양한 방법을 통해 구축될 수 있으며, 이에 대한 구체적인 방법은 Sambrook et al.(2001), Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press에 개시되어 있으며, 이 문헌은 본 명세서에 참조로서 삽입된다.
본 발명의 벡터는 전형적으로 클로닝을 위한 벡터 또는 발현을 위한 벡터로서 구축될 수 있다. 또한, 본 발명의 벡터는 원핵 세포 또는 진핵 세포를 숙주로하여 구축될 수 있다. 본 발명의 벡터가 발현 벡터이고, 원핵 세포를 숙주로 하는 경우에는, 전사를 진행시킬 수 있는 강력한 프로모터 (예컨대, tac 프로모터, lac 프로모터, lacUV5 프로모터, lpp 프로모터, pLλ 프로모터, pRλ 프로모터, rac5 프로모터, amp 프로모터, recA 프로모터, SP6 프로모터, trp 프로모터 및 T7 프로모터 등), 해독의 개시를 위한 라이보좀 결합 자리 및 전사/해독 종결 서열을 포함하는 것이 일반적이다. 숙주 세포로서 E. coli (예컨대, HB101, BL21, DH5α 등)가 이용되는 경우, E. coli 트립토판 생합성 경로의 프로모터 및 오퍼레이터 부위 (Yanofsky, C.(1984), J. Bacteriol., 158:1018-1024) 그리고 파아지 λ의 좌향 프로모터 (pLλ 프로모터, Herskowitz, I. and Hagen, D.(1980), Ann. Rev. Genet., 14:399-445)가 조절 부위로서 이용될 수 있다.
레트로바이러스 (Retrovirus)는 숙주세포의 게놈에 통합기능이 있으며, 인체에 해가 없지만 통합 시 정상세포의 기능을 억제할 수 있고, 다양한 세포에 감염되고, 증식이 쉬우며, 1~7 kb 정도의 외부 유전자를 수용할 수 있고, 복제결핍 바이러스를 생성할 수 있는 능력이 있다는 특징을 갖는다. 그러나 레트로바이러스는 유사분열 이후의 세포에 감염되기 힘들며, in vivo 상태에서 유전자 전달이 어렵고, 체세포 조직을 항시 in vitro에서 증식하여야만 한다는 단점도 지니고 있다. 또한, 레트로바이러스는 원형암유전자 (proto-oncogene)에 통합될 수 있어 돌연변이의 위험성이 있고 세포를 괴사시킬 수도 있다.
한편, 아데노바이러스 (Adenovirus)는 클로닝 벡터 (cloning vector)로 여러가지 장점을 가지는데, 중간정도의 크기로 세포 핵 속에서 복제될 수 있으며, 임상적으로 무독성이고, 외부 유전자를 삽입하여도 안정적이며, 유전자의 재배열이나 손실이 일어나지 않고, 진핵생물을 형질전환시킬 수 있으며, 숙주세포 염색체에 통합되어도 안정적이면서도 높은 수준으로 발현된다. 아데노바이러스의 좋은 숙주세포는 인간의 조혈, 림프, 골수종의 원인이 되는 세포이다. 그러나 선상 DNA라서 증식이 어렵고, 감염된 바이러스를 회복시키는 것이 쉽지 않으며, 바이러스의 감염율이 낮다. 또한 전달된 유전자의 발현이 1~2주 후에 가장 많이 되고, 일부 세포에서는 3~4주 정도만 발현이 유지된다. 또한, 문제가 되는 것은 높은 면역 항원성을 갖는다는 것이다.
아데노-조력 바이러스 (Adeno-associated virus, AAV)는 상기와 같은 문제점들을 보완할 수 있으면서, 유전자 치료제로의 많은 장점을 가지고 있어 최근에 선호되고 있다. 아는 아데노-위성 바이러스 (adeno-satellitovirus)라고도 불린다. 아데노-조력 바이러스 입자의 직경은 20 nm이며, 인체에 해가 거의 없는 것으로 알려져 유럽에서 유전자 치료제로 판매가 승인되기도 하였다.
상기 아데노-조력 바이러스는 단일가닥의 원바이러스 (Provirus)로서, 복제하기 위하여 보조 바이러스를 필요로 하고, 아데노-조력 바이러스 게놈 (genome)은 4,680 bp로서 감염세포의 염색체 19번 특정부위에 삽입이 가능하다. 트랜스 유전자 (trans-gene)는 각각 145bp의 두 개의 역위말단반복 (inverted terminal repeat, ITR) 서열부분과 시그날 서열 (signal sequence)부분에 의해 연결된 플라즈미드 DNA (plasmid DNA)에 삽입된다. 아데노-조력 바이러스 rep 부분과 cap 부분을 발현시키는 다른 플라즈미드 DNA와 함께 트랜스펙션 (transfection)시키고 아데노 바이러스는 보조 바이러스로서 첨가한다. 아데노-조력 바이러스는 유전자를 전달하는 숙주세포의 범위가 넓고, 반복투여 시 면역 부작용이 적으며, 유전자 발현 기간이 긴 장점을 지니고 있다. 더구나, 아데노-조력 바이러스 게놈이 숙주세포의 염색체에 통합되어도 안전하고, 숙주의 유전자발현을 변형시키거나 재배열시키지 않는다.
상기 아데노-조력 바이러스는 4종의 혈청형이 있는 것으로 알려져 있다. 목적 유전자의 전달에 사용될 수 있는 많은 아데노-조력 바이러스의 혈청형들 중에서, 가장 널리 연구된 벡터는 아데노-조력 바이러스 혈청형 2 (Adeno-associated virus serotype 2)로, 낭포성섬유증, 혈우병 및 카나반 병의 임상적 유전자 전달에 현재 사용되고 있다. 또한, 최근에는 암 유전자 치료 분야 (cancer gene therapy)에서 rAAV (recombinant adeno-associated virus)의 잠재성이 증가하고 있다. 본 발명 에서 사용한 것 역시 아데노-조력 바이러스 혈청형 2이며, 적절한 바이러스벡터를 선택하여 적용할 수 있으나, 이에 제한되지 않는다.
또한, 본 발명의 벡터에서 외래 유전자를 발현시키는 경우, 진핵 세포를 숙주로 하는 경우에는, 포유동물 세포의 유전체로부터 유래된 프로모터 (예: 메탈로티오닌 프로모터) 또는 포유동물 바이러스로부터 유래된 프로모터 (예: 아데노바이러스 후기 프로모터, 백시니아 바이러스 7.5K 프로모터, SV40 프로모터, 사이토메갈로바이러스 프로모터 및 HSV의 사 프로모터)가 이용될 수 있으며, 구체적으로, 레트로바이러스의 LTR 프로모터, 사이토메갈로바이러스 CMV 프로모터, 라우스 육종 바이러스 RSV 프로모터, MMT 프로모터, EF-1 알파 프로모터, UB6 프로모터, 치킨 베타-액틴 프로모터, CAG 프로모터, RPE65 프로모터 및 옵신 프로모터로 이루어진 군으로부터 선택된 프로모터로 이루어진 군에서 선택된 1종 이상을 포함할 수 있으나, 이에 제한되지 않으며, 전사 종결 서열로서 폴리아데닐화 서열을 일반적으로 갖는다.
본 발명의 벡터는 단백질의 정제를 용이하게 하기 위하여, 필요에 따라 다른 서열과 융합될 수도 있으며, 융합되는 서열은 예컨대, 글루타티온 S-트랜스퍼라제 (Pharmacia, USA), 말토스 결합 단백질 (NEB, USA), FLAG (IBI, USA) 및 6x His (hexahistidine; Quiagen, USA) 등이 이용될 수 있으나, 이에 제한되지는 않는다. 또한, 본 발명의 발현 벡터는 선택표지로서, 당업계에서 통상적으로 이용되는 항생제 내성 유전자를 포함할 수 있으며, 예를 들어 암피실린, 겐타마이신, 카베니실린, 클로람페니콜, 스트렙토마이신, 카나마이신, 게네티신, 네오마이신 및 테트라사이클린에 대한 내성 유전자가 있다.
본 발명에서 이용된 바이러스는 또한 상기 기술된 것외의 다른 변형을 또한 포함할 수 있다. 어떠한 추가의 성분 또는 변형도 임의로 사용할 수 있지만 본 발명을 위해 의무적이지는 않다.
외인성 성분의 삽입은 바이러스의 효과를 향상시킬 수 있다. 외인성 조직 또는 종양-특이적인 프로모터의 사용은 재조합체 벡터에서 일반적이며 이들은 또한 본 발명에서 이용될 수 있다.
일 측면에서, 본 발명은 본 발명의 바이러스 벡터를 포함하는, 폐암세포 검출용 조성물에 관한 것이다.
일 구현예에서, 폐암세포는 순환종양세포일 수 있으며, 혈중 순환종양세포인 것이 가장 바람직하다.
본 발명에서 사용된 용어, "폐암세포 검출용"은 "폐암암세포 동정용"과 서로 교차 또는 혼용되어 사용될 수 있다.
본 발명에서 사용된 용어 "검출하다", "검출하는" 또는 "검출"은 검출가능하게 표지된 조성물의 발견 또는 식별 또는 특이적 관찰의 일반적 실시를 기재하는 것일 수 있다.
일 측면에서, 본 발명은 본 발명의 폐암세포 검출용 조성물을 포함하는 폐암 진단용 키트에 관한 것이다.
일 구현예에서, 상기 키트는 대상체 또는 환자로부터 생체 시료를 수집하기 위한 도구 및/또는 시약 뿐 아니라 그 시료로부터 세포를 분리하기 위한 도구 및/또는 시약을 더 포함할 수 있다.
일 구현예에서, 폐암세포는 순환종양세포일 수 있으며, 혈중 순환종양세포인 것이 가장 바람직하다.
일 측면에서, 본 발명은 개체로부터 분리된 시료에 본 발명의 폐암세포 검출용 조성물을 처리하는 단계를 포함하는, 폐암세포 검출 방법에 관한 것이다.
일 구현예에서, 상기 방법은 시료 내 표지 유전자 발현 세포가 폐암세포인 것으로 판단하는 단계를 추가로 포함할 수 있다.
일 구현예에서, 시료는 혈액, 혈청, 혈장, 타액, 복수, 누액, 뇌척수액, 흉수, 활액, 림프, 또는 뇨인, 암세포 검출 방법.혈액, 양수, 혈장, 혈청, 정액, 골수, 뇨, 조직 생검 또는 세포로부터 단백질을 추출한 용액일 수 있으며, 혈액인 것이 가장 바람직하다.
일 구현예에서, 폐암세포는 순환종양세포일 수 있으며, 혈중 순환종양세포인 것이 가장 바람직하다.
본 발명의 폐암세포 검출 방법은 개체의 혈액 내에 존재하는 폐암세포를 검출 및 분리 동정하는 방법을 포함한다.
일 측면에서, 본 발명은 개체로부터 분리된 혈액에 본 발명의 폐암세포 검출용 조성물을 처리하는 단계; 혈액 내 표지 유전자 발현 세포를 분리하는 단계; 및 분리한 세포를 배양하는 단계를 포함하는, 순환폐암세포의 분리 및 배양 방법에 관한 것이다.
일 측면에서, 본 발명은 개체로부터 분리된 혈액에 본 발명의 폐암세포 검출용 조성물을 처리하는 단계; 혈액 내 표지 유전자 발현 세포를 분리하는 단계; 및 분리한 세포를 배양하는 단계를 포함하는, 폐암의 진단에 필요한 정보를 제공하는 방법에 관한 것이다.
본 발명에서 사용되는 용어, "개체"란, 암이 발병하였거나 발병할 수 있는 인간을 포함한 원숭이, 소, 말, 양, 돼지, 닭, 칠면조, 메추라기, 고양이, 개, 마우스, 쥐, 토끼 또는 기니아 피그를 포함한 모든 동물을 의미한다.
이하, 실시예를 통하여 본 발명을 보다 자세히 설명한다. 다만, 상기 실시예 및 실험예는 본 발명에 대한 예시로 제시되는 것으로, 당업자에게 주지 저명한 기술 또는 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우에는 그 상세한 설명을 생략할 수 있고, 이에 의해 본 발명이 제한되지는 않는다. 본 발명은 후술하는 특허청구범위의 기재 및 그로부터 해석되는 균등 범주 내에서 다양한 변형 및 응용이 가능하다.
실시예 1. 폐암 특이적 발현의 아데노바이러스 벡터 제조
1-1. 프라이머 제작
본 발명의 재조합 벡터에 사용할 SFTPA2 (Surfactant Protein A2) 유전자의 프로모터 또는 SFTPC (Surfactant Protein C) 유전자의 프로모터를 클로닝하기 위한 PCR을 수행하기 위해 표 1과 같이 SpeⅠ엔자임 서열을 포함한 프라이머를 제작하였다.
염기서열 서열번호
hSFTPA2 SpeⅠ 정방향(F) GCGCACTAGTACTCCTCAATGGTTTGGGAT 1
hSFTPA2 SpeⅠ 역방향(R) ATATACTAGTGCTTCAGGCCTCCGGCTGGA 2
hSFTPC SpeⅠ 정방향(F) GCGCACTAGTATTGGCAGATTTGACCTCGT 3
hSFTPC SpeⅠ 역방향(R) GTGTACTAGTCTTGCTGCAGGTGCTATGCT 4
1-2. 아데노바이러스 벡터 제조
OD260 Inc. adenoquick 2.0 cosmid 제작 제품을 이용하여 진행하였으며, 본 발명자의 실험실 내에서 보유 중인 pAd1127-htert와 pAd1129-GFP를 이용하여 하기와 같은 실험을 진행하였다. (1) pAd1127-htert의 htert 프로모터를 상기 실시예 1.1의 프라이머를 이용하여 PCR 수행 결과 얻은 SFTPA2 유전자 프로모터 또는 SFTPC 유전자 프로모터로 교체 하였다 (pAd1127-SFTPA2promoter, pAd1127-SFTPCpromoter). (2) SFTPA2 유전자 프로모터 또는 SFTPC 유전자 프로모터, E1a 및 E1b를 포함하는 pAd1127-PSApromoter 벡터 또는 pAd1127-SFTPCpromoter 벡터; E2, L1, L2, L3 및 L4를 포함하는 pAd1128 벡터; E3, fiber 및 GFP를 포함하는 pAd1129-GFP 벡터; 및 E4를 포함하는 pAd1130 벡터를 이용하여 코스미드 벡터를 제작하였다(도 1 참조). (3) miR-142를 표적하는 서열의 클로닝 방법은 miR-142-3p 표적서열을 BbsⅠ제한효소 사이트를 추가하여 합성하고, pA1129 벡터와 상기 BbsⅠ제한효소 사이트를 포함하여 합성한 miR-142-3p 표적서열을 BbsⅠ으로 엔자임 커팅 (enzyme cutting) 후 라이게이션 (ligation)을 하여 최종 코스미드 벡터를 제작하였다 (도 2 참조). 이때, miR-142-3p 표적서열이 벡터에 들어간 자리는 pA1129 벡터 GFP 유전자의 5’UTR 지역이다.
상기 (2)에서 제작한 코스미드 벡터의 1) CMV 프로모터; 2) 상기 프로모터에 작동 가능하게 연결된 목적 단백질 (표지 단백질)을 코딩하는 염기서열; 및 3) 상기 염기서열의 5’UTR에 삽입된 miR-142를 표적화하는 염기서열을 포함하는 벡터를 제조하였다. 이때 CMV 프로모터는 예시일 뿐이며 레트로바이러스의 LTR 프로모터, 사이토메갈로바이러스 CMV 프로모터, 라우스 육종 바이러스 RSV 프로모터, MMT 프로모터, EF-1 알파 프로모터, UB6 프로모터, 치킨 베타-액틴 프로모터, CAG 프로모터, RPE65 프로모터 및 옵신 프로모터로 이루어진 군으로부터 선택된 프로모터로 이루어진 군에서 선택된 1종 을 포함할 수 있으나, 이에 제한되지 않는다.
실시예 2. 폐암세포 특이적 발현 확인
(1) 293A 세포 6 well에 접종 (seeding)하였다. 이때의 cell confluency는 70% 정도였다. (2) 다음날 cacl2 0.5M 100ul에 PacI digested cosmid 2ug가 들어간 용액과 2X Hebs 용액 100ul를 혼합하였고, 이 때 Hebs 용액에 cacl2 및 DNA 용액을 조금씩 넣어주면서 vortexing 하였다. 이 후, 15분 incubation 후 293A 세포에 한 방울씩 뿌렸다. (3) 배지 교체 (media change)를 6시간 후 및 overnight 두 가지 방법 모두 진행하여 수행하였다. (4) cell confluency가 100% 정도 되었을때 60파이 dish로 옮겨주었다. (5) 그 후, 매일 배지 (media)를 1ml씩 첨가하였고, (6) 약 5일 후, 배양상등액 (sup)과 세포를 추출하여, 3번 동결과 융해 (Freeze and thaw) 하였다. (7) 293A 세포에 바이러스를 감염 (infection) 시켜서 round up 진행하였고, 이의 결과를 도 3에 나타내었다.
도 3에서 나타난 바와 같이, 재조합 벡터의 암세포 (293A) 특이적 발현 효과를 확인하였다.
따라서 본 발명의 아데노바이러스 벡터는 폐암 세포에 높은 특이적 발현성을 가짐을 확인하여 이를 폐암 진단용 키트, 폐암 진단용 조성물, 폐암세포를 검출하는 방법 및 폐암을 진단하는 방법에 활용할 수 있음을 확인하였다.
이제까지 본 발명에 대하여 그 바람직한 실시 예들을 중심으로 살펴보았다.
본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시 예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허청구 범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.
<110> University-Industry Cooperation Group of Kyung Hee University Core Pharma Co.,Ltd <120> Lung cancer diagnostic composition comprising an adenovirus vector and use thereof <130> PN1909-438 <160> 9 <170> KoPatentIn 3.0 <210> 1 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of SFTPA2 promoter gene <400> 1 ggcgactagt acattgtttg ctgcacgttg 30 <210> 2 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of SFTPA2 promoter gene <400> 2 atatactagt gcttcaggcc tccggctgga 30 <210> 3 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of SFTPC promoter gene <400> 3 gcgcactagt attggcagat ttgacctcgt 30 <210> 4 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of SFTPC promoter gene <400> 4 gtgtactagt cttgctgcag gtgctatgct 30 <210> 5 <211> 1691 <212> DNA <213> Artificial Sequence <220> <223> SFTPA2 promoter gene <400> 5 actcctcaat ggtttgggat ttttcaaacc aattccaaat aagccagggg agattttcct 60 gtctgggttc ccatcattac cacctccaca gttggagatt ccctcctaat ccagatgctc 120 aaaccaggac cctaacaact agggcttcat atatcactgg gacacttagc ataatcacca 180 ttgcaggttg ccagacatca cccaaagtga tgttctacca aagacagaaa ctgcagctct 240 cccatgacct tccaggtgct gccctgactc ctccagctga acctgctagg gcagagggaa 300 gtggaacgtg cagcatattt gcctgccact ctgtccatgt gcccaccaac tcggctgcag 360 gctccagaga cactggctgg gggtcctggg gctcagagct gagcaaggtt acagggcaga 420 caggttggcc tggatcaacc tgaccaacct cagctgaagc agtgacttct tcattgtctg 480 aaactaaacc aggaacccag tggagctccc tgagtgcaga gtgggtgacc ttagccagtg 540 aggcatttct cttcttacag acctggtatt tttctttacc aggttctgtg ctcccctcaa 600 gggtcctaag tattcctgca gcctgagtgt tctgcaggga aatgtgctgt gtaaacacta 660 tgcctatttc ctgcttggag aacaggtttt gtggtagagc tctcaggggt ggggaagaag 720 cctggcagcc cacatgctat aaattctgtt gtccaccttt atgctctaac ttggaggcag 780 agacccaagc agctggaggc tctgtgtgtg ggtgagttta gccacatccc ctaggtgttc 840 tccagcttga ggatcgcagg cagagaggac cagcccagca gccacaggcc tgaccaaggc 900 ccaggttggg aaggagggca actccccatt ttccactggg aggtgtttca cagcatggtc 960 aacatggatg acctacaaag accctcatgc tttgatatct tccttggcca gatccatccc 1020 taccagggac tcagcagggc ctaagcaggg ggagtgggag aacagacccc aagaggaagg 1080 aagacctcat cactgactgt gagagggtca tcaccccccg atgatcacct tgaagggtcc 1140 aggctgtggg ccctattcat ctcttttcat tctcaggtcg ctgatttctt ggagcctgaa 1200 aagaaggtaa ctgggcatat gagggacaga tggagtgagt cagtgagtga gtgacctcat 1260 ccttccttct aacagcctgg gagggacagg gcaggttttc tgcagagcat ggaagattca 1320 gctgaaatca gagaggtgaa gccagtttcc cagggtaaca tagtgaggca ctgaaagaaa 1380 ggagactgca ctggagccca ggtccctggg ctccccagag ctccttactc ttcctcctcc 1440 tcagcagtct tgagacccca caacctccag ccggaggcct gaagcatgag gccatgccag 1500 gtgccaggtg atgcttggaa ttttcccggg tgcttcgggt cttcccagca ctctggtctc 1560 gcccgccctg cctctcgggc tctgcccagc ttcctgagtc ctgacagagc acagtggggg 1620 agatgttggc agaggtggca gatgggctca cggccatccc tccggcagga gcagcgactg 1680 gacccagagc c 1691 <210> 6 <211> 1051 <212> DNA <213> Artificial Sequence <220> <223> SFTPC promoter gene <400> 6 attggcagat ttgacctcgt aaatacatag agatggcttt gggaaggcac taggaaagac 60 agagaaaaga gaaggagaca gtcctcaaag ctgatcgtat ttgggtgagt attattctca 120 gggcaaattt aggatcagag gatgcagaaa ggggagtcta gaggggtaga gtgtagacca 180 cagggtgagt gagctgattc gaggatgggg agactgggag cccaccagtg accagagcca 240 gccctgttca gggctgtccg ggcagaagaa agcagtgtca gacctggaat ctgccatcag 300 cacagcctgc aattgacaga caagcccaga gcaaagaagg aagcactgca catgagtaag 360 agcttgccac cagtggggac agagtttcca gaattaggaa aataatcact gggggcaagt 420 ttgaggttgg taccagatat gtgggaggag gcaaggtaag ggaaagagta cttgaagttg 480 gaactggtcc ttgcagggaa atgcacattt atgaaacccc gaaaactgat gtcaaagcac 540 ctcctgcctt gggcagagtc ctctcagagt ctacaggtgc tgcctccaga accctcttcc 600 tggagcgcat ccctatgtat ctagaaattc tgctgggaaa tatgatggtc agacccttgg 660 ccacctgaaa ggttcagggt ggtagaagaa aaaggaaagc cacagggcag caggggcagg 720 tgccagcaag gaaggcaggc acgccaggaa gacacccatg gtgagaagtg cagatggccc 780 gagggcaagt ttgctcaact cacccaggtt tgctcttgct ggggccaaga ggactcatgt 840 gccagggcca agggcccttg ggggctctca cagggggctt atctgggctt cggttctgga 900 gggccaggaa caaacaggct tcaaagccaa gggcttggct ggcacacagg gggcttggtc 960 cttcacctct gtcccctctc cctacggaca catataagac cctggtcaca cctgggagag 1020 gaggagagga gagcatagca cctgcagcaa g 1051 <210> 7 <211> 132 <212> DNA <213> Artificial Sequence <220> <223> miR-142-3p target sequence <400> 7 ccagaagctt gctagctcca taaagtagga aacactacat cactccataa agtaggaaac 60 actacagata tctccataaa gtaggaaaca ctacatcact ccataaagta ggaaacacta 120 caaagcttgt ag 132 <210> 8 <211> 46408 <212> DNA <213> Artificial Sequence <220> <223> recombinant vector sequence(SFTPA2-GFP-miR142) <400> 8 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatatttgt ctagggagct 360 agcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tggctagcac 420 tagtactcct caatggtttg ggatttttca aaccaattcc aaataagcca ggggagattt 480 tcctgtctgg gttcccatca ttaccacctc cacagttgga gattccctcc taatccagat 540 gctcaaacca ggaccctaac aactagggct tcatatatca ctgggacact tagcataatc 600 accattgcag gttgccagac atcacccaaa gtgatgttct accaaagaca gaaactgcag 660 ctctcccatg accttccagg tgctgccctg actcctccag ctgaacctgc tagggcagag 720 ggaagtggaa cgtgcagcat atttgcctgc cactctgtcc atgtgcccac caactcggct 780 gcaggctcca gagacactgg ctgggggtcc tggggctcag agctgagcaa ggttacaggg 840 cagacaggtt ggcctggatc aacctgacca acctcagctg aagcagtgac ttcttcattg 900 tctgaaacta aaccaggaac ccagtggagc tccctgagtg cagagtgggt gaccttagcc 960 agtgaggcat ttctcttctt acagacctgg tatttttctt taccaggttc tgtgctcccc 1020 tcaagggtcc taagtattcc tgcagcctga gtgttctgca gggaaatgtg ctgtgtaaac 1080 actatgccta tttcctgctt ggagaacagg ttttgtggta gagctctcag gggtggggaa 1140 gaagcctggc agcccacatg ctataaattc tgttgtccac ctttatgctc taacttggag 1200 gcagagaccc aagcagctgg aggctctgtg tgtgggtgag tttagccaca tcccctaggt 1260 gttctccagc ttgaggatcg caggcagaga ggaccagccc agcagccaca ggcctgacca 1320 aggcccaggt tgggaaggag ggcaactccc cattttccac tgggaggtgt ttcacagcat 1380 ggtcaacatg gatgacctac aaagaccctc atgctttgat atcttccttg gccagatcca 1440 tccctaccag ggactcagca gggcctaagc agggggagtg ggagaacaga ccccaagagg 1500 aaggaagacc tcatcactga ctgtgagagg gtcatcaccc cccgatgatc accttgaagg 1560 gtccaggctg tgggccctat tcatctcttt tcattctcag gtcgctgatt tcttggagcc 1620 tgaaaagaag gtaactgggc atatgaggga cagatggagt gagtcagtga gtgagtgacc 1680 tcatccttcc ttctaacagc ctgggaggga cagggcaggt tttctgcaga gcatggaaga 1740 ttcagctgaa atcagagagg tgaagccagt ttcccagggt aacatagtga ggcactgaaa 1800 gaaaggagac tgcactggag cccaggtccc tgggctcccc agagctcctt actcttcctc 1860 ctcctcagca gtcttgagac cccacaacct ccagccggag gcctgaagca tgaggccatg 1920 ccaggtgcca ggtgatgctt ggaattttcc cgggtgcttc gggtcttccc agcactctgg 1980 tctcgcccgc cctgcctctc gggctctgcc cagcttcctg agtcctgaca gagcacagtg 2040 ggggagatgt tggcagaggt ggcagatggg ctcacggcca tccctccggc aggagcagcg 2100 actggaccca gagccactag tacaccggga ctgaaaatga gacatattat ctgccacgga 2160 ggtgttatta ccgaagaaat ggccgccagt cttttggacc agctgatcga agaggtactg 2220 gctgataatc ttccacctcc tagccatttt gaaccaccta cccttcacga actgtatgat 2280 ttagacgtga cggcccccga agatcccaac gaggaggcgg tttcgcagat ttttcccgac 2340 tctgtaatgt tggcggtgca ggaagggatt gacttactca cttttccgcc ggcgcccggt 2400 tctccggagc cgcctcacct ttcccggcag cccgagcagc cggagcagag agccttgggt 2460 ccggtttcta tgccaaacct tgtaccggag gtgatcgatc ttacctgcca cgaggctggc 2520 tttccaccca gtgacgacga ggatgaagag ggtgaggagt ttgtgttaga ttatgtggag 2580 caccccgggc acggttgcag gtcttgtcat tatcaccgga ggaatacggg ggacccagat 2640 attatgtgtt cgctttgcta tatgaggacc tgtggcatgt ttgtctacag tcctgtgtct 2700 gaacctgagc ctgagcccga gccagaaccg gagcctgcaa gacctacccg ccgtcctaaa 2760 atggcgcctg ctatcctgag acgcccgaca tcacctgtgt ctagagaatg caatagtagt 2820 acggatagct gtgactccgg tccttctaac acacctcctg agatacaccc ggtggtcccg 2880 ctgtgcccca ttaaaccagt tgccgtgaga gttggtgggc gtcgccaggc tgtggaatgt 2940 atcgaggact tgcttaacga gcctgggcaa cctttggact tgagctgtaa acgccccagg 3000 ccataaggtg taaacctgtg acccctctcc ctcccccccc taacgttact ggccgaagcc 3060 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt 3120 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc 3180 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc 3240 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc 3300 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg 3360 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct 3420 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat 3480 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc 3540 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc 3600 acaacccggg atcctctaga gtcgacatgg aggcttggga gtgtttggaa gatttttctg 3660 ctgtgcgtaa cttgctggaa cagagctcta acagtacctc ttggttttgg aggtttctgt 3720 ggggctcatc ccaggcaaag ttagtctgca gaattaagga ggattacaag tgggaatttg 3780 aagagctttt gaaatcctgt ggtgagctgt ttgattcttt gaatctgggt caccaggcgc 3840 ttttccaaga gaaggtcatc aagactttgg atttttccac accggggcgc gctgcggctg 3900 ctgttgcttt tttgagtttt ataaaggata aatggagcga agaaacccat ctgagcgggg 3960 ggtacctgct ggattttctg gccatgcatc tgtggagagc ggttgtgaga cacaagaatc 4020 gcctgctact gttgtcttcc gtccgcccgg cgataatacc gacggaggag cagcagcagc 4080 agcaggagga agccaggcgg cggcggcagg agcagagccc atggaacccg agagccggcc 4140 tggaccctcg ggaatgaatg ttgtacaggt ggctgaactg tatccagaac tgagacgcat 4200 tttgacaatt acagaggatg ggcaggggct aaagggggta aagagggagc ggggggcttg 4260 tgaggctaca gaggaggcta ggaatctagc ttttagctta atgaccagac accgtcctga 4320 gtgtattact tttcaacaga tcaaggataa ttgcgctaat gagcttgatc tgctggcgca 4380 gaagtattcc atagagcagc tgaccactta ctggctgcag ccaggggatg attttgagga 4440 ggctattagg gtatatgcaa aggtggcact taggccagat tgcaagtaca agatcagcaa 4500 acttgtaaat atcaggaatt gttgctacat ttctgggaac ggggccgagg tggagataga 4560 tacggaggat agggtggcct ttagatgtag catgataaat atgtggccgg gggtgcttgg 4620 catggacggg gtggttatta tgaatgtaag gtttactggc cccaatttta gcggtacggt 4680 tttcctggcc aataccaacc ttatcctaca cggtgtaagc ttctatgggt ttaacaatac 4740 ctgtgtggaa gcctggaccg atgtaagggt tcggggctgt gccttttact gctgctggaa 4800 gggggtggtg tgtcgcccca aaagcagggc ttcaattaag aaatgcctct ttgaaaggtg 4860 taccttgggt atcctgtctg agggtaactc cagggtgcgc cacaatgtgg cctccgactg 4920 tggttgcttc atgctagtga aaagcgtggc tgtgattaag cataacatgg tatgtggcaa 4980 ctgcgaggac agggcctctc agatgctgac ctgctcggac ggcaactgtc acctgctgaa 5040 gaccattcac gtagccagcc actctcgcaa ggcctggcca gtgtttgagc ataacatact 5100 gacccgctgt tccttgcatt tgggtaacag gaggggggtg ttcctacctt accaatgcaa 5160 tttgagtcac actaagatat tgcttgagcc cgagagcatg tccaaggtga acctgaacgg 5220 ggtgtttgac atgaccatga agatctggaa ggtgctgagg tacgatgaga cccgcaccag 5280 gtgcagaccc tgcgagtgtg gcggtaaaca tattaggaac cagcctgtga tgctggatgt 5340 gaccgaggag ctgaggcccg atcacttggt gctggcctgc acccgcgctg agtttggctc 5400 tagcgatgaa gatacagatt gaagtactga aatgtgtggg cgtggcttaa gggtgggaaa 5460 gaatatataa ggtgggggtc ttatgtagtt ttgtatctgt tttgcagcag ccgccgccgc 5520 catgagcacc aactcgtttg atggaagcat tgtgagctca tatttgacaa cgcgcatgcc 5580 cccatgggcc ggggtgcgtc agaatgtgat gggctccagc attgatggtc gccccgtcct 5640 gcccgcaaac tctactacct tgacctacga gaccgtgtct ggaacgccgt tggagactgc 5700 agcctccgcc gccgcttcag ccgctgcagc caccgcccgc gggattgtga ctgactttgc 5760 tttcctgagc ccgcttgcaa gcagtgcagc ttcccgttca tccgcccgcg atgacaagtt 5820 gacggctctt ttggcacaat tggattcttt gacccgggaa cttaatgtcg tttctcagca 5880 gctgttggat ctgcgccagc aggtttctgc cctgaaggct tcctcccctc ccaatgcggt 5940 ttaacggtcc gggccagagt ggccaacata aataaaaaac cagactctgt ttggatttgg 6000 atcaagcaag tgtcttgctg tctttattta ggggttttgc gcgcgcggta ggcccgggac 6060 cagcggtctc ggtcgttgag ggtcctgtgt attttttcca ggacgtggta aaggtgactc 6120 tggatgttca gatacatggg cataagcccg tctctggggt ggaggtagca ccactgcaga 6180 gcttcatgct gcggggtggt gttgtagatg atccagtcgt agcaggagcg ctgggcgtgg 6240 tgcctaaaaa tgtctttcag tagcaagctg attgccaggg gcaggccctt ggtgtaagtg 6300 tttacaaagc ggttaagctg ggatgggtgc atacgtgggg atatgagatg catcttggac 6360 tgtattttta ggttggctat gttcccagcc atatccctcc ggggattcat gttgtgcaga 6420 accaccagca cagtgtatcc ggtgcacttg ggaaatttgt catgtagctt agaaggaaat 6480 gcgtggaaga acttggagac gcccttgtga cctccaagat tttccatgca ttcgtccata 6540 atgatggcaa tgggcccacg ggcggcggcc tgggcgaaga tatttctggg atcactaacg 6600 tcatagttgt gttccaggat gagatcgtca taggccattt ttacaaagcg cgggcggagg 6660 gtgccagact gcggtataat ggttccatcc ggcccagggg cgtagttacc ctcacagatt 6720 tgcatttccc acgctttgag ttcagatggg gggatcatgt ctacctgcgg ggcgatgaag 6780 aaaacggttt ccggggtagg ggagatcagc tgggaagaaa gcaggttcct gagcagctgc 6840 gacttaccgc agccggtggg cccgtaaatc acacctatta ccggctgcaa ctggtagtta 6900 agagagctgc agctgccgtc atccctgagc aggggggcca cttcgttaag catgtccctg 6960 actcgcatgt tttccctgac caaatccgcc agaaggcgct cgccgcccag cgatagcagt 7020 tcttgcaagg aagcaaagtt tttcaacggt ttgagaccgt ccgccgtagg catgcttttg 7080 agcgtttgac caagcagttc caggcggtcc cacagctcgg tcacctgctc tacggcatct 7140 cgatccagca tatctcctcg tttcgcgggt tggggcggct ttcgctgtac ggcagtagtc 7200 ggtgctcgtc cagacgggcc agggtcatgt ctttccacgg gcgcagggtc ctcgtcagcg 7260 tagtctgggt cacggtgaag gggtgcgctc cgggctgcgc gctggccagg gtgcgcttga 7320 ggctggtcct gctggtgctg aagcgctgcc ggtcttcgcc ctgcgcgtcg gccaggtagc 7380 atttgaccat ggtgtcatag tccagcccct ccgcggcgtg gcccttggcg cgcagcttgc 7440 ccttggagga ggcgccgcac gaggggcagt gcagactttt gagggcgtag agcttgggcg 7500 cgagaaatac cgattccggg gagtaggcat ccgcgccgca ggccccgcag acggtctcgc 7560 attccacgag ccaggtgagc tctggccgtt cggggtcaaa aaccaggttt cccccatgct 7620 ttttgatgcg tttcttacct ctggtttcca tgagccggtg tccacgctcg gtgacgaaaa 7680 ggctgtccgt gtccccgtat acagacttga gaggcctgtc ctcgagcggt gttccgcggt 7740 cctcctcgta tagaaactcg gaccactctg agacaaaggc tcgcgtccag gccagcacga 7800 aggaggctaa gtgggagggg tagcggtcgt tgtccactag ggggtccact cgctccaggg 7860 tgtgaagaca catgtcgccc tcttcggcat caaggaaggt gattggtttg taggtgtagg 7920 ccacgtgacc gggtgttcct gaaggggggc tataaaaggg ggtgggggcg cgttcgtcct 7980 cactctcttc cgcatcgctg tctgcgaggg ccagctgttg gggtgagtac tccctctgaa 8040 aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat 8100 tcacctggcc cgcggtgatg cctttgaggg tggccgcatc catctggtca gaaaagacaa 8160 tctttttgtt gtcaagcttg gtggcaaacg acccgtagag ggcgttggac agcaacttgg 8220 cgatggagcg cagggtttgg tttttgtcgc gatcggcgcg ctccttggcc gcgatgttta 8280 gctgcacgta ttcgcgcgca acgcaccgcc attcgggaaa gacggtggtg cgctcgtcgg 8340 gcaccaggtg cacgcgccaa ccgcggttgt gcagggtgac aaggtcaacg ctggtggcta 8400 cctctccgcg taggcgctcg ttggtccagc agaggcggcc gcccttgcgc gagcagaatg 8460 gcggtagggg gtctagctgc gtctcgtccg gggggtctgc gtccacggta aagaccccgg 8520 gcagcaggcg cgcgtcgaag tagtctatct tgcatccttg caagtctagc gcctgctgcc 8580 atgcgcgggc ggcaagcgcg cgctcgtatg ggttgagtgg gggaccccat ggcatggggt 8640 gggtgagcgc ggaggcgtac atgccgcaaa tgtcgtaaac gtagaggggc tctctgagta 8700 ttccaagata tgtagggtag catcttccac cgcggatgct ggcgcgcacg taatcgtata 8760 gttcgtgcga gggagcgagg aggtcgggac cgaggttgct acgggcgggc tgctctgctc 8820 ggaagactat ctgcctgaag atggcatgtg agttggatga tatggttgga cgctggaaga 8880 cgttgaagct ggcgtctgtg agacctaccg cgtcacgcac gaaggaggcg taggagtcgc 8940 gcagcttgtt gaccagctcg gcggtgacct gcacgtctag ggcgcagtag tccagggttt 9000 ccttgatgat gtcatactta tcctgtccct tttttttcca cagctcgcgg ttgaggacaa 9060 actcttcgcg gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtaag 9120 agcctagcat gtagaactgg ttgacggcct ggtaggcgca gcatcccttt tctacgggta 9180 gcgcgtatgc ctgcgcggcc ttccggagcg aggtgtgggt gagcgcaaag gtgtccctga 9240 ccatgacttt gaggtactgg tatttgaagt cagtgtcgtc gcatccgccc tgctcccaga 9300 gcaaaaagtc cgtgcgcttt ttggaacgcg gatttggcag ggcgaaggtg acatcgttga 9360 agagtatctt tcccgcgcga ggcataaagt tgcgtgtgat gcggaagggt cccggcacct 9420 cggaacggtt gttaattacc tgggcggcga gcacgatctc gtcaaagccg ttgatgttgt 9480 ggcccacaat gtaaagttcc aagaagcgcg ggatgccctt gatggaaggc aattttttaa 9540 gttcctcgta ggtgagctct tcaggggagc tgagcccgtg ctctgaaagg gcccagtctg 9600 caagatgagg gttggaagcg acgaatgagc tccacaggtc acgggccatt agcatttgca 9660 ggtggtcgcg aaaggtccta aactggcgac ctatggccat tttttctggg gtgatgcagt 9720 agaaggtaag cgggtcttgt tcccagcggt cccatccaag gttcgcggct aggtctcgcg 9780 cggcagtcac tagaggctca tctccgccga acttcatgac cagcatgaag ggcacgagct 9840 gcttcccaaa ggcccccatc caagtatagg tctctacatc gtaggtgaca aagagacgct 9900 cggtgcgagg atgcgagccg atcgggaaga actggatctc ccgccaccaa ttggaggagt 9960 ggctattgat gtggtgaaag tagaagtccc tgcgacgggc cgaacactcg tgctggcttt 10020 tgtaaaaacg tgcgcagtac tggcagcggt gcacgggctg tacatcctgc acgaggttga 10080 cctgacgacc gcgcacaagg aagcagagtg ggaatttgag cccctcgcct ggcgggtttg 10140 gctggtggtc ttctacttcg gctgcttgtc cttgaccgtc tggctgctcg aggggagtta 10200 cggtggatcg gaccaccacg ccgcgcgagc ccaaagtcca gatgtccgcg cgcggcggtc 10260 ggagcttgat gacaacatcg cgcagatggg agctgtccat ggtctggagc tcccgcggcg 10320 tcaggtcagg cgggagctcc tgcaggttta cctcgcatag acgggtcagg gcgcgggcta 10380 gatccaggtg atacctaatt tccaggggct ggttggtggc ggcgtcgatg gcttgcaaga 10440 ggccgcatcc ccgcggcgcg actacggtac cgcgcggcgg gcggtgggcc gcgggggtgt 10500 ccttggatga tgcatctaaa agcggtgacg cgggcgagcc cccggaggta gggggggctc 10560 cggacccgcc gggagagggg gcaggggcac gtcggcgccg cgcgcgggca ggagctggtg 10620 ctgcgcgcgt aggttgctgg cgaacgcgac gacgcggcgg ttgatctcct gaatctggcg 10680 cctctgcgtg aagacgacgg gcccggtgag cttgaacctg aaagagagtt cgacagaatc 10740 aatttcggtg tcgttgacgg cggcctggcg caaaatctcc tgcacgtctc ctgagttgtc 10800 ttgataggcg atctcggcca tgaactgctc gatctcttcc tcctggagat ctccgcgtcc 10860 ggctcgctcc acggtggcgg cgaggtcgtt ggaaatgcgg gccatgagct gcgagaaggc 10920 gttgaggcct ccctcgttcc agacgcggct gtagaccacg cccccttcgg catcgcgggc 10980 gcgcatgacc acctgcgcga gattgagctc cacgtgccgg gcgaagacgg cgtagtttcg 11040 caggcgctga aagaggtagt tgagggtggt ggcggtgtgt tctgccacga agaagtacat 11100 aacccagcgt cgcaacgtgg attcgttgat atcccccaag gcctcaaggc gctccatggc 11160 ctcgtagaag tccacggcga agttgaaaaa ctgggagttg cgcgccgaca cggttaactc 11220 ctcctccaga agacggatga gctcggcgac agtgtcgcgc acctcgcgct caaaggctac 11280 aggggcctct tcttcttctt caatctcctc ttccataagg gcctcccctt cttcttcttc 11340 tggcggcggt gggggagggg ggacacggcg gcgacgacgg cgcaccggga ggcggtcgac 11400 aaagcgctcg atcatctccc cgcggcgacg gcgcatggtc tcggtgacgg cgcggccgtt 11460 ctcgcggggg cgcagttgga agacgccgcc cgtcatgtcc cggttatggg ttggcggggg 11520 gctgccatgc ggcagggata cggcgctaac gatgcatctc aacaattgtt gtgtaggtac 11580 tccgccgccg agggacctga gcgagtccgc atcgaccgga tcggaaaacc tctcgagaaa 11640 ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc gtggcgggcg gcagcgggcg 11700 gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg taattaaagt aggcggtctt 11760 gagacggcgg atggtcgaca gaagcaccat gtccttgggt ccggcctgct gaatgcgcag 11820 gcggtcggcc atgccccagg cttcgttttg acatcggcgc aggtctttgt agtagtcttg 11880 catgagcctt tctaccggca cttcttcttc tccttcctct tgtcctgcat ctcttgcatc 11940 tatcgctgcg gcggcggcgg agtttggccg taggtggcgc cctcttcctc ccatgcgtgt 12000 gaccccgaag cccctcatcg gctgaagcag ggctaggtcg gcgacaacgc gctcggctaa 12060 tatggcctgc tgcacctgcg tgagggtaga ctggaagtca tccatgtcca caaagcggtg 12120 gtatgcgccc gtgttgatgg tgtaagtgca gttggccata acggaccagt taacggtctg 12180 gtgacccggc tgcgagagct cggtgtacct gagacgcgag taagccctcg agtcaaatac 12240 gtagtcgttg caagtccgca ccaggtactg gtatcccacc aaaaagtgcg gcggcggctg 12300 gcggtagagg ggccagcgta gggtggccgg ggctccgggg gcgagatctt ccaacataag 12360 gcgatgatat ccgtagatgt acctggacat ccaggtgatg ccggcggcgg tggtggaggc 12420 gcgcggaaag tcgcggacgc ggttccagat gttgcgcagc ggcaaaaagt gctccatggt 12480 cgggacgctc tggccggtca ggcgcgcgca atcgttgacg ctctagaccg tgcaaaagga 12540 gagcctgtaa gcgggcactc ttccgtggtc tggtggataa attcgcaagg gtatcatggc 12600 ggacgaccgg ggttcgagcc ccgtatccgg ccgtccgccg tgatccatgc ggttaccgcc 12660 cgcgtgtcga acccaggtgt gcgacgtcag acaacggggg agtgctcctt ttggcttcct 12720 tccaggcgcg gcggctgctg cgctagcttt tttggccact ggccgcgcgc agcgtaagcg 12780 gttaggctgg aaagcgaaag cattaagtgg ctcgctccct gtagccggag ggttattttc 12840 caagggttga gtcgcgggac ccccggttcg agtctcggac cggccggact gcggcgaacg 12900 ggggtttgcc tccccgtcat gcaagacccc gcttgcaaat tcctccggaa acagggacga 12960 gccccttttt tgcttttccc agatgcatcc ggtgctgcgg cagatgcgcc cccctcctca 13020 gcagcggcaa gagcaagagc agcggcagac atgcagggca ccctcccctc ctcctaccgc 13080 gtcaggaggg gcgacatccg cggttgacgc ggcagcagat ggtgattacg aacccccgcg 13140 gcgccgggcc cggcactacc tggacttgga ggagggcgag ggcctggcgc ggctaggagc 13200 gccctctcct gagcggcacc caagggtgca gctgaagcgt gatacgcgtg aggcgtacgt 13260 gccgcggcag aacctgtttc gcgaccgcga gggagaggag cccgaggaga tgcgggatcg 13320 aaagttccac gcagggcgcg agctgcggca tggcctgaat cgcgagcggt tgctgcgcga 13380 ggaggacttt gagcccgacg cgcgaaccgg gattagtccc gcgcgcgcac acgtggcggc 13440 cgccgacctg gtaaccgcat acgagcagac ggtgaaccag gagattaact ttcaaaaaag 13500 ctttaacaac cacgtgcgta cgcttgtggc gcgcgaggag gtggctatag gactgatgca 13560 tctgtgggac tttgtaagcg cgctggagca aaacccaaat agcaagccgc tcatggcgca 13620 gctgttcctt atagtgcagc acagcaggga caacgaggca ttcagggatg cgctgctaaa 13680 catagtagag cccgagggcc gctggctgct cgatttgata aacatcctgc agagcatagt 13740 ggtgcaggag cgcagcttga gcctggctga caaggtggcc gccatcaact attccatgct 13800 tagcctgggc aagttttacg cccgcaagat ataccatacc ccttacgttc ccatagacaa 13860 ggaggtaaag atcgaggggt tctacatgcg catggcgctg aaggtgctta ccttgagcga 13920 cgacctgggc gtttatcgca acgagcgcat ccacaaggcc gtgagcgtga gccggcggcg 13980 cgagctcagc gaccgcgagc tgatgcacag cctgcaaagg gccctggctg gcacgggcag 14040 cggcgataga gaggccgagt cctactttga cgcgggcgct gacctgcgct gggccccaag 14100 ccgacgcgcc ctggaggcag ctggggccgg acctgggctg gcggtggcac ccgcgcgcgc 14160 tggcaacgtc ggcggcgtgg aggaatatga cgaggacgat gagtacgagc cagaggacgg 14220 cgagtactaa gcggtgatgt ttctgatcag atgatgcaag acgcaacgga cccggcggtg 14280 cgggcggcgc tgcagagcca gccgtccggc cttaactcca cggacgactg gcgccaggtc 14340 atggaccgca tcatgtcgct gactgcgcgc aatcctgacg cgttccggca gcagccgcag 14400 gccaaccggc tctccgcaat tctggaagcg gtggtcccgg cgcgcgcaaa ccccacgcac 14460 gagaaggtgc tggcgatcgt aaacgcgctg gccgaaaaca gggccatccg gcccgacgag 14520 gccggcctgg tctacgacgc gctgcttcag cgcgtggctc gttacaacag cggcaacgtg 14580 cagaccaacc tggaccggct ggtgggggat gtgcgcgagg ccgtggcgca gcgtgagcgc 14640 gcgcagcagc agggcaacct gggctccatg gttgcactaa acgccttcct gagtacacag 14700 cccgccaacg tgccgcgggg acaggaggac tacaccaact ttgtgagcgc actgcggcta 14760 atggtgactg agacaccgca aagtgaggtg taccagtctg ggccagacta ttttttccag 14820 accagtagac aaggcctgca gaccgtaaac ctgagccagg ctttcaaaaa cttgcagggg 14880 ctgtgggggg tgcgggctcc cacaggcgac cgcgcgaccg tgtctagctt gctgacgccc 14940 aactcgcgcc tgttgctgct gctaatagcg cccttcacgg acagtggcag cgtgtcccgg 15000 gacacatacc taggtcactt gctgacactg taccgcgagg ccataggtca ggcgcatgtg 15060 gacgagcata ctttccagga gattacaagt gtcagccgcg cgctggggca ggaggacacg 15120 ggcagcctgg aggcaaccct aaactacctg ctgaccaacc ggcggcagaa gatcccctcg 15180 ttgcacagtt taaacagcga ggaggagcgc attttgcgct acgtgcagca gagcgtgagc 15240 cttaacctga tgcgcgacgg ggtaacgccc agcgtggcgc tggacatgac cgcgcgcaac 15300 atggaaccgg gcatgtatgc ctcaaaccgg ccgtttatca accgcctaat ggactacttg 15360 catcgcgcgg ccgccgtgaa ccccgagtat ttcaccaatg ccatcttgaa cccgcactgg 15420 ctaccgcccc ctggtttcta caccggggga ttcgaggtgc ccgagggtaa cgatggattc 15480 ctctgggacg acatagacga cagcgtgttt tccccgcaac cgcagaccct gctagagttg 15540 caacagcgcg agcaggcaga ggcggcgctg cgaaaggaaa gcttccgcag gccaagcagc 15600 ttgtccgatc taggcgctgc ggccccgcgg tcagatgcta gtagcccatt tccaagcttg 15660 atagggtctc ttaccagcac tcgcaccacc cgcccgcgcc tgctgggcga ggaggagtac 15720 ctaaacaact cgctgctgca gccgcagcgc gaaaaaaacc tgcctccggc atttcccaac 15780 aacgggatag agagcctagt ggacaagatg agtagatgga agacgtacgc gcaggagcac 15840 agggacgtgc caggcccgcg cccgcccacc cgtcgtcaaa ggcacgaccg tcagcggggt 15900 ctggtgtggg aggacgatga ctcggcagac gacagcagcg tcctggattt gggagggagt 15960 ggcaacccgt ttgcgcacct tcgccccagg ctggggagaa tgttttaaaa aaaaaaaaaa 16020 gcatgatgca aaataaaaaa ctcaccaagg ccatggcacc gagcgttggt tttcttgtat 16080 tccccttagt atgcggcgcg cggcgatgta tgaggaaggt cctcctccct cctacgagag 16140 tgtggtgagc gcggcgccag tggcggcggc gctgggttct cccttcgatg ctcccctgga 16200 cccgccgttt gtgcctccgc ggtacctgcg gcctaccggg gggagaaaca gcatccgtta 16260 ctctgagttg gcacccctat tcgacaccac ccgtgtgtac ctggtggaca acaagtcaac 16320 ggatgtggca tccctgaact accagaacga ccacagcaac tttctgacca cggtcattca 16380 aaacaatgac tacagcccgg gggaggcaag cacacagacc atcaatcttg acgaccggtc 16440 gcactggggc ggcgacctga aaaccatcct gcataccaac atgccaaatg tgaacgagtt 16500 catgtttacc aataagttta aggcgcgggt gatggtgtcg cgcttgccta ctaaggacaa 16560 tcaggtggag ctgaaatacg agtgggtgga gttcacgctg cccgagggca actactccga 16620 gaccatgacc atagacctta tgaacaacgc gatcgtggag cactacttga aagtgggcag 16680 acagaacggg gttctggaaa gcgacatcgg ggtaaagttt gacacccgca acttcagact 16740 ggggtttgac cccgtcactg gtcttgtcat gcctggggta tatacaaacg aagccttcca 16800 tccagacatc attttgctgc caggatgcgg ggtggacttc acccacagcc gcctgagcaa 16860 cttgttgggc atccgcaagc ggcaaccctt ccaggagggc tttaggatca cctacgatga 16920 tctggagggt ggtaacattc ccgcactgtt ggatgtggac gcctaccagg cgagcttgaa 16980 agatgacacc gaacagggcg ggggtggcgc aggcggcagc aacagcagtg gcagcggcgc 17040 ggaagagaac tccaacgcgg cagccgcggc aatgcagccg gtggaggaca tgaacgatca 17100 tgccattcgc ggcgacacct ttgccacacg ggctgaggag aagcgcgctg aggccgaagc 17160 agcggccgaa gctgccgccc ccgctgcgca acccgaggtc gagaagcctc agaagaaacc 17220 ggtgatcaaa cccctgacag aggacagcaa gaaacgcagt tacaacctaa taagcaatga 17280 cagcaccttc acccagtacc gcagctggta ccttgcatac aactacggcg accctcagac 17340 cggaatccgc tcatggaccc tgctttgcac tcctgacgta acctgcggct cggagcaggt 17400 ctactggtcg ttgccagaca tgatgcaaga ccccgtgacc ttccgctcca cgcgccagat 17460 cagcaacttt ccggtggtgg gcgccgagct gttgcccgtg cactccaaga gcttctacaa 17520 cgaccaggcc gtctactccc aactcatccg ccagtttacc tctctgaccc acgtgttcaa 17580 tcgctttccc gagaaccaga ttttggcgcg cccgccagcc cccaccatca ccaccgtcag 17640 tgaaaacgtt cctgctctca cagatcacgg gacgctaccg ctgcgcaaca gcatcggagg 17700 agtccagcga gtgaccatta ctgacgccag acgccgcacc tgcccctacg tttacaaggc 17760 cctgggcata gtctcgccgc gcgtcctatc gagccgcact ttttgagcaa gcatgtccat 17820 ccttatatcg cccagcaata acacaggctg gggcctgcgc ttcccaagca agatgtttgg 17880 cggggccaag aagcgctccg accaacaccc agtgcgcgtg cgcgggcact accgcgcgcc 17940 ctggggcgcg cacaaacgcg gccgcactgg gcgcaccacc gtcgatgacg ccatcgacgc 18000 ggtggtggag gaggcgcgca actacacgcc cacgccgcca ccagtgtcca cagtggacgc 18060 ggccattcag accgtggtgc gcggagcccg gcgctatgct aaaatgaaga gacggcggag 18120 gcgcgtagca cgtcgccacc gccgccgacc cggcactgcc gcccaacgcg cggcggcggc 18180 cctgcttaac cgcgcacgtc gcaccggccg acgggcggcc atgcgagcag ctcgaaggct 18240 ggccgcgggt attgtcactg tgccccccag gtccaggcga cgagcggccg ccgcagcagc 18300 cgcggccatt agtgctatga ctcagggtcg caggggcaac gtgtattggg tgcgcgactc 18360 ggttagcggc ctgcgcgtgc ccgtgcgcac ccgccccccg cgcaactaga ttgcaagaaa 18420 aaactactta gactcgtact gttgtatgta tccagcggcg gcggcgcgca acgaagctat 18480 gtccaagcgc aaaatcaaag aagagatgct ccaggtcatc gcgccggaga tctatggccc 18540 cccgaagaag gaagagcagg attacaagcc ccgaaagcta aagcgggtca aaaagaaaaa 18600 gaaagatgat gatgatgaac ttgacgacga ggtggaactg ctgcacgcta ccgcgcccag 18660 gcgacgggta cagtggaaag gtcgacgcgt aaaacgtgtt ttgcgacccg gcaccaccgt 18720 agtctttacg cccggtgagc gctccacccg cacctacaag cgcgtgtatg atgaggtgta 18780 cggcgacgag gacctgcttg agcaggccaa cgagcgcctc ggggagtttg cctacggaaa 18840 gcggcataag gacatgctgg cgttgccgct ggacgagggc aacccaacac ctagcctaaa 18900 gcccgtaaca ctgcagcagg tgctgcccgc gcttgcaccg tccgaagaaa agcgcggcct 18960 aaagcgcgag tctggtgact tggcacccac cgtgcagctg atggtaccca agcgccagcg 19020 actggaagat gtcttggaaa aaatgaccgt ggaacctggg ctggagcccg aggtccgcgt 19080 gcggccaatc aagcaggtgg cgccgggact gggcgtgcag accgtggacg ttcagatacc 19140 cactaccagt agcaccagta ttgccaccgc cacagagggc atggagacac aaacgtcccc 19200 ggttgcctca gcggtggcgg atgccgcggt gcaggcggtc gctgcggccg cgtccaagac 19260 ctctacggag gtgcaaacgg acccgtggat gtttcgcgtt tcagcccccc ggcgcccgcg 19320 ccgttcgagg aagtacggcg ccgccagcgc gctactgccc gaatatgccc tacatccttc 19380 cattgcgcct acccccggct atcgtggcta cacctaccgc cccagaagac gagcaactac 19440 ccgacgccga accaccactg gaacccgccg ccgccgtcgc cgtcgccagc ccgtgctggc 19500 cccgatttcc gtgcgcaggg tggctcgcga aggaggcagg accctggtgc tgccaacagc 19560 gcgctaccac cccagcatcg tttaaaagcc ggtctttgtg gttcttgcag atatggccct 19620 cacctgccgc ctccgtttcc cggtgccggg attccgagga agaatgcacc gtaggagggg 19680 catggccggc cacggcctga cgggcggcat gcgtcgtgcg caccaccggc ggcggcgcgc 19740 gtcgcaccgt cgcatgcgcg gcggtatcct gcccctcctt attccactga tcgccgcggc 19800 gattggcgcc gtgcccggaa ttgcatccgt ggccttgcag gcgcagagac actgattaaa 19860 aacaagttgc atgtggaaaa atcaaaataa aaagtctgga ctctcacgct cgcttggtcc 19920 tgtaactatt ttgtagaatg gaagacatca actttgcgtc tctggccccg cgacacggct 19980 cgcgcccgtt catgggaaac tggcaagata tcggcaccag caatatgagc ggtggcgcct 20040 tcagctgggg ctcgctgtgg agcggcatta aaaatttcgg ttccaccgtt aagaactatg 20100 gcagcaaggc ctggaacagc agcacaggcc agatgctgag ggataagttg aaagagcaaa 20160 atttccaaca aaaggtggta gatggcctgg cctctggcat tagcggggtg gtggacctgg 20220 ccaaccaggc agtgcaaaat aagattaaca gtaagcttga tccccgccct cccgtagagg 20280 agcctccacc ggccgtggag acagtgtctc cagaggggcg tggcgaaaag cgtccgcgcc 20340 ccgacaggga agaaactctg gtgacgcaaa tagacgagcc tccctcgtac gaggaggcac 20400 taaagcaagg cctgcccacc acccgtccca tcgcgcccat ggctaccgga gtgctgggcc 20460 agcacacacc cgtaacgctg gacctgcctc cccccgccga cacccagcag aaacctgtgc 20520 tgccaggccc gaccgccgtt gttgtaaccc gtcctagccg cgcgtccctg cgccgcgccg 20580 ccagcggtcc gcgatcgttg cggcccgtag ccagtggcaa ctggcaaagc acactgaaca 20640 gcatcgtggg tctgggggtg caatccctga agcgccgacg atgcttctga tagctaacgt 20700 gtcgtatgtg tgtcatgtat gcgtccatgt cgccgccaga ggagctgctg agccgccgcg 20760 cgcccgcttt ccaagatggc taccccttcg atgatgccgc agtggtctta catgcacatc 20820 tcgggccagg acgcctcgga gtacctgagc cccgggctgg tgcagtttgc ccgcgccacc 20880 gagacgtact tcagcctgaa taacaagttt agaaacccca cggtggcgcc tacgcacgac 20940 gtgaccacag accggtccca gcgtttgacg ctgcggttca tccctgtgga ccgtgaggat 21000 actgcgtact cgtacaaggc gcggttcacc ctagctgtgg gtgataaccg tgtgctggac 21060 atggcttcca cgtactttga catccgcggc gtgctggaca ggggccctac ttttaagccc 21120 tactctggca ctgcctacaa cgccctggct cccaagggtg ccccaaatcc ttgcgaatgg 21180 gatgaagctg ctactgctct tgaaataaac ctagaagaag aggacgatga caacgaagac 21240 gaagtagacg agcaagctga gcagcaaaaa actcacgtat ttgggcaggc gccttattct 21300 ggtataaata ttacaaagga gggtattcaa ataggtgtcg aaggtcaaac acctaaatat 21360 gccgataaaa catttcaacc tgaacctcaa ataggagaat ctcagtggta cgaaacagaa 21420 attaatcatg cagctgggag agtcctaaaa aagactaccc caatgaaacc atgttacggt 21480 tcatatgcaa aacccacaaa tgaaaatgga gggcaaggca ttcttgtaaa gcaacaaaat 21540 ggaaagctag aaagtcaagt ggaaatgcaa tttttctcaa ctactgaggc agccgcaggc 21600 aatggtgata acttgactcc taaagtggta ttgtacagtg aagatgtaga tatagaaacc 21660 ccagacactc atatttctta catgcccact attaaggaag gtaactcacg agaactaatg 21720 ggccaacaat ctatgcccaa caggcctaat tacattgctt ttagggacaa ttttattggt 21780 ctaatgtatt acaacagcac gggtaatatg ggtgttctgg cgggccaagc atcgcagttg 21840 aatgctgttg tagatttgca agacagaaac acagagcttt cataccagct tttgcttgat 21900 tccattggtg atagaaccag gtacttttct atgtggaatc aggctgttga cagctatgat 21960 ccagatgtta gaattattga aaatcatgga actgaagatg aacttccaaa ttactgcttt 22020 ccactgggag gtgtgattaa tacagagact cttaccaagg taaaacctaa aacaggtcag 22080 gaaaatggat gggaaaaaga tgctacagaa ttttcagata aaaatgaaat aagagttgga 22140 aataattttg ccatggaaat caatctaaat gccaacctgt ggagaaattt cctgtactcc 22200 aacatagcgc tgtatttgcc cgacaagcta aagtacagtc cttccaacgt aaaaatttct 22260 gataacccaa acacctacga ctacatgaac aagcgagtgg tggctcccgg gctagtggac 22320 tgctacatta accttggagc acgctggtcc cttgactata tggacaacgt caacccattt 22380 aaccaccacc gcaatgctgg cctgcgctac cgctcaatgt tgctgggcaa tggtcgctat 22440 gtgcccttcc acatccaggt gcctcagaag ttctttgcca ttaaaaacct ccttctcctg 22500 ccgggctcat acacctacga gtggaacttc aggaaggatg ttaacatggt tctgcagagc 22560 tccctaggaa atgacctaag ggttgacgga gccagcatta agtttgatag catttgcctt 22620 tacgccacct tcttccccat ggcccacaac accgcctcca cgcttgaggc catgcttaga 22680 aacgacacca acgaccagtc ctttaacgac tatctctccg ccgccaacat gctctaccct 22740 atacccgcca acgctaccaa cgtgcccata tccatcccct cccgcaactg ggcggctttc 22800 cgcggctggg ccttcacgcg ccttaagact aaggaaaccc catcactggg ctcgggctac 22860 gacccttatt acacctactc tggctctata ccctacctag atggaacctt ttacctcaac 22920 cacaccttta agaaggtggc cattaccttt gactcttctg tcagctggcc tggcaatgac 22980 cgcctgctta cccccaacga gtttgaaatt aagcgctcag ttgacgggga gggttacaac 23040 gttgcccagt gtaacatgac caaagactgg ttcctggtac aaatgctagc taactataac 23100 attggctacc agggcttcta tatcccagag agctacaagg accgcatgta ctccttcttt 23160 agaaacttcc agcccatgag ccgtcaggtg gtggatgata ctaaatacaa ggactaccaa 23220 caggtgggca tcctacacca acacaacaac tctggatttg ttggctacct tgcccccacc 23280 atgcgcgaag gacaggccta ccctgctaac ttcccctatc cgcttatagg caagaccgca 23340 gttgacagca ttacccagaa aaagtttctt tgcgatcgca ccctttggcg catcccattc 23400 tccagtaact ttatgtccat gggcgcactc acagacctgg gccaaaacct tctctacgcc 23460 aactccgccc acgcgctaga catgactttt gaggtggatc ccatggacga gcccaccctt 23520 ctttatgttt tgtttgaagt ctttgacgtg gtccgtgtgc accagccgca ccgcggcgtc 23580 atcgaaaccg tgtacctgcg cacgcccttc tcggccggca acgccacaac ataaagaagc 23640 aagcaacatc aacaacagct gccgccatgg gctccagtga gcaggaactg aaagccattg 23700 tcaaagatct tggttgtggg ccatattttt tgggcaccta tgacaagcgc tttccaggct 23760 ttgtttctcc acacaagctc gcctgcgcca tagtcaatac ggccggtcgc gagactgggg 23820 gcgtacactg gatggccttt gcctggaacc cgcactcaaa aacatgctac ctctttgagc 23880 cctttggctt ttctgaccag cgactcaagc aggtttacca gtttgagtac gagtcactcc 23940 tgcgccgtag cgccattgct tcttcccccg accgctgtat aacgctggaa aagtccaccc 24000 aaagcgtaca ggggcccaac tcggccgcct gtggactatt ctgctgcatg tttctccacg 24060 cctttgccaa ctggccccaa actcccatgg atcacaaccc caccatgaac cttattaccg 24120 gggtacccaa ctccatgctc aacagtcccc aggtacagcc caccctgcgt cgcaaccagg 24180 aacagctcta cagcttcctg gagcgccact cgccctactt ccgcagccac agtgcgcaga 24240 ttaggagcgc cacttctttt tgtcacttga aaaacatgta aaaataatgt actagagaca 24300 ctttcaataa aggcaaatgc ttttatttgt acactctcgg gtgattattt acccccaccc 24360 ttgccgtctg cgccgtttaa aaatcaaagg ggttctgccg cgcatcgcta tgcgccactg 24420 gcagggacac gttgcgatac tggtgtttag tgctccactt aaactcaggc acaaccatcc 24480 gcggcagctc ggtgaagttt tcactccaca ggctgcgcac catcaccaac gcgtttagca 24540 ggtcgggcgc cgatatcttg aagtcgcagt tggggcctcc gccctgcgcg cgcgagttgc 24600 gatacacagg gttgcagcac tggaacacta tcagcgccgg gtggtgcacg ctggccagca 24660 cgctcttgtc ggagatcaga tccgcgtcca ggtcctccgc gttgctcagg gcgaacggag 24720 tcaactttgg tagctgcctt cccaaaaagg gcgcgtgccc aggctttgag ttgcactcgc 24780 accgtagtgg catcaaaagg tgaccgtgcc cggtctgggc gttaggatac agcgcctgca 24840 taaaagcctt gatctgctta aaagccacct gagcctttgc gccttcagag aagaacatgc 24900 cgcaagactt gccggaaaac tgattggccg gacacgcggc gtcgtgcacg cagcaccttg 24960 cgtcggtgtt ggagatctgc accacatttc ggccccaccg gttcttcacg atcttggcct 25020 tgctagactg ctccttcagc gcgcgctgcc cgttttcgct cgtcacatcc atttcaatca 25080 cgtgctcctt atttatcata atgcttccgt gtagacactt aagctcgcct tcgatctcag 25140 cgcagcggtg cagccacaac gcgcagcccg tgggctcgtg atgcttgtag gtcacctctg 25200 caaacgactg caggtacgcc tgcaggaatc gccccatcat cgtcacaaag gtcttgttgc 25260 tggtgaaggt cagctgcaac ccgcggtgct cctcgttcag ccaggtcttg catacggccg 25320 ccagagcttc cacttggtca ggcagtagtt tgaagttcgc ctttagatcg ttatccacgt 25380 ggtacttgtc catcagcgcg cgcgcagcct ccatgccctt ctcccacgca gacacgatcg 25440 gcacactcag cgggttcatc accgtaattt cactttccgc ttcgctgggc tcttcctctt 25500 cctcttgcgt ccgcatacca cgcgccactg ggtcgtcttc attcagccgc cgcactgtgc 25560 gcttacctcc tttgccatgc ttgattagca ccggtgggtt gctgaaaccc accatttgta 25620 gcgccacatc ttctctttct tcctcgctgt ccacgattac ctctggtgat ggcgggcgct 25680 cgggcttggg agaagggcgc ttctttttct tcttgggcgc aatggccaaa tccgccgccg 25740 aggtcgatgg ccgcgggctg ggtgtgcgcg gcaccagcgc gtcttgtgat gagtcttcct 25800 cgtcctcgga ctcgatacgc cgcctcatcc gcttttttgg gggcgcccgg ggaggcggcg 25860 gcgacgggga cggggacgac acgtcctcca tggttggggg acgtcgcgcc gcaccgcgtc 25920 cgcgctcggg ggtggtttcg cgctgctcct cttcccgact ggccatttcc ttctcctata 25980 ggcagaaaaa gatcatggag tcagtcgaga agaaggacag cctaaccgcc ccctctgagt 26040 tcgccaccac cgcctccacc gatgccgcca acgcgcctac caccttcccc gtcgaggcac 26100 ccccgcttga ggaggaggaa gtgattatcg agcaggaccc aggttttgta agcgaagacg 26160 acgaggaccg ctcagtacca acagaggata aaaagcaaga ccaggacaac gcagaggcaa 26220 acgaggaaca agtcgggcgg ggggacgaaa ggcatggcga ctacctagat gtgggagacg 26280 acgtgctgtt gaagcatctg cagcgccagt gcgccattat ctgcgacgcg ttgcaagagc 26340 gcagcgatgt gcccctcgcc atagcggatg tcagccttgc ctacgaacgc cacctattct 26400 caccgcgcgt accccccaaa cgccaagaaa acggcacatg cgagcccaac ccgcgcctca 26460 acttctaccc cgtatttgcc gtgccagagg tgcttgccac ctatcacatc tttttccaaa 26520 actgcaagat acccctatcc tgccgtgcca accgcagccg agcggacaag cagctggcct 26580 tgcggcaggg cgctgtcata cctgatatcg cctcgctcaa cgaagtgcca aaaatctttg 26640 agggtcttgg acgcgacgag aagcgcgcgg caaacgctct gcaacaggaa aacagcgaaa 26700 atgaaagtca ctctggagtg ttggtggaac tcgagggtga caacgcgcgc ctagccgtac 26760 taaaacgcag catcgaggtc acccactttg cctacccggc acttaaccta ccccccaagg 26820 tcatgagcac agtcatgagt gagctgatcg tgcgccgtgc gcagcccctg gagagggatg 26880 caaatttgca agaacaaaca gaggagggcc tacccgcagt tggcgacgag cagctagcgc 26940 gctggcttca aacgcgcgag cctgccgact tggaggagcg acgcaaacta atgatggccg 27000 cagtgctcgt taccgtggag cttgagtgca tgcagcggtt ctttgctgac ccggagatgc 27060 agcgcaagct agaggaaaca ttgcactaca cctttcgaca gggctacgta cgccaggcct 27120 gcaagatctc caacgtggag ctctgcaacc tggtctccta ccttggaatt ttgcacgaaa 27180 accgccttgg gcaaaacgtg cttcattcca cgctcaaggg cgaggcgcgc cgcgactacg 27240 tccgcgactg cgtttactta tttctatgct acacctggca gacggccatg ggcgtttggc 27300 agcagtgctt ggaggagtgc aacctcaagg agctgcagaa actgctaaag caaaacttga 27360 aggacctatg gacggccttc aacgagcgct ccgtggccgc gcacctggcg gacatcattt 27420 tccccgaacg cctgcttaaa accctgcaac agggtctgcc agacttcacc agtcaaagca 27480 tgttgcagaa ctttaggaac tttatcctag agcgctcagg aatcttgccc gccacctgct 27540 gtgcacttcc tagcgacttt gtgcccatta agtaccgcga atgccctccg ccgctttggg 27600 gccactgcta ccttctgcag ctagccaact accttgccta ccactctgac ataatggaag 27660 acgtgagcgg tgacggtcta ctggagtgtc actgtcgctg caacctatgc accccgcacc 27720 gctccctggt ttgcaattcg cagctgctta acgaaagtca aattatcggt acctttgagc 27780 tgcagggtcc ctcgcctgac gaaaagtccg cggctccggg gttgaaactc actccggggc 27840 tgtggacgtc ggcttacctt cgcaaatttg tacctgagga ctaccacgcc cacgagatta 27900 ggttctacga agaccaatcc cgcccgccta atgcggagct taccgcctgc gtcattaccc 27960 agggccacat tcttggccaa ttgcaagcca tcaacaaagc ccgccaagag tttctgctac 28020 gaaagggacg gggggtttac ttggaccccc agtccggcga ggagctcaac ccaatccccc 28080 cgccgccgca gccctatcag cagcagccgc gggcccttgc ttcccaggat ggcacccaaa 28140 aagaagctgc agctgccgcc gccacccacg gacgaggagg aatactggga cagtcaggca 28200 gaggaggttt tggacgagga ggaggaggac atgatggaag actgggagag cctagacgag 28260 gaagcttccg aggtcgaaga ggtgtcagac gaaacaccgt caccctcggt cgcattcccc 28320 tcgccggcgc cccagaaatc ggcaaccggt tccagcatgg ctacaacctc cgctcctcag 28380 gcgccgccgg cactgcccgt tcgccgaccc aaccgtagat gggacaccac tggaaccagg 28440 gccggtaagt ccaagcagcc gccgccgtta gcccaagagc aacaacagcg ccaaggctac 28500 cgctcatggc gcgggcacaa gaacgccata gttgcttgct tgcaagactg tgggggcaac 28560 atctccttcg cccgccgctt tcttctctac catcacggcg tggccttccc ccgtaacatc 28620 ctgcattact accgtcatct ctacagccca tactgcaccg gcggcagcgg cagcaacagc 28680 agcggccaca cagaagcaaa ggcgaccgga tagcaagact ctgacaaagc ccaagaaatc 28740 cacagcggcg gcagcagcag gaggaggagc gctgcgtctg gcgcccaacg aacccgtatc 28800 gacccgcgag cttagaaaca ggatttttcc cactctgtat gctatatttc aacagagcag 28860 gggccaagaa caagagctga aaataaaaaa caggtctctg cgatccctca cccgcagctg 28920 cctgtatcac aaaagcgaag atcagcttcg gcgcacgctg gaagacgcgg aggctctctt 28980 cagtaaatac tgcgcgctga ctcttaagga ctagtttcgc gccctttctc aaatttaagc 29040 gcgaaaacta cgtcatctcc agcggccaca cccggcgcca gcacctgttg tcagcgccat 29100 tatgagcaag gaaattccca cgccctacat gtggagttac cagccacaaa tgggacttgc 29160 ggctggagct gcccaagact actcaacccg aataaactac atgagcgcgg gaccccacat 29220 gatatcccgg gtcaacggaa tacgcgccca ccgaaaccga attctcctgg aacaggcggc 29280 tattaccacc acacctcgta ataaccttaa tccccgtagt tggcccgctg ccctggtgta 29340 ccaggaaagt cccgctccca ccactgtggt acttcccaga gacgcccagg ccgaagttca 29400 gatgactaac tcaggggcgc agcttgcggg cggctttcgt cacagggtgc ggtcgcccgg 29460 gcagggtata actcacctga caatcagagg gcgaggtatt cagctcaacg acgagtcggt 29520 gagctcctcg cttggtctcc gtccggacgg gacatttcag atcggcggcg ccggccgctc 29580 ttcattcacg cctcgtcagg caatcctaac tctgcagacc tcgtcctctg agccgcgctc 29640 tggaggcatt ggaactctgc aatttattga ggagtttgtg ccatcggtct actttaaccc 29700 cttctcggga cctcccggcc actatccgga tcaatttatt cctaactttg acgcggtaaa 29760 ggactcggcg gacggctacg actgaggcca atgaggccat gttaagtgga gaggcagagc 29820 aactgcgcct gaaacacctg gtccactgtc gccgccacaa gtgctttgcc cgcgactccg 29880 gtgagttttg ctactttgaa ttgcccgagg atcatatcga gggcccggcg cacggcgtcc 29940 ggcttaccgc ccagggagag cttgcccgta gcctgattcg ggagtttacc cagcgccccc 30000 tgctagttga gcgggacagg ggaccctgtg ttctcactgt gatttgcaac tgtcctaacc 30060 ctggattaca tcaagatctc ctaggctagc actagtcgtt acataactta cggtaaatgg 30120 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 30180 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 30240 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 30300 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 30360 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 30420 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 30480 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa 30540 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 30600 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca 30660 tagaagacac cggcccagaa gcttgctagc tccataaagt aggaaacact acatcactcc 30720 ataaagtagg aaacactaca gatatctcca taaagtagga aacactacat cactccataa 30780 agtaggaaac actacaaagc ttgtagcggc cgccaccatg actgccctga ccgaaggtgc 30840 taagctgttt gagaaggaga ttccgtacat caccgagctg gaaggggacg tcgaaggaat 30900 gaagttcatc atcaagggag aaggaaccgg ggacgctacg actggaacca ttaaggccaa 30960 gtatatctgt accactggag atctgccagt gccttgggcc acccttgtgt caaccctctc 31020 gtatggagtg cagtgttttg ctaagtaccc tagccacatt aaggacttct tcaaatccgc 31080 catgccggaa ggttataccc aagagcgcac catttctttt gagggagatg gagtgtacaa 31140 gacccgcgcg atggtcacct atgagagggg atctatctac aaccgggtga ctctgactgg 31200 agaaaacttt aagaaggacg ggcatattct tcggaagaat gtcgccttcc agtgccctcc 31260 cagcatcctt tacattctcc ccgacactgt gaacaacgga atccgcgtgg agttcaatca 31320 agcctacgac atcgaggggg tgacggagaa gctggtgacc aagtgtagcc agatgaatcg 31380 gccactggcc ggttcagcgg ctgtccacat tccgcgctac catcatatca cttatcacac 31440 taagctctcc aaagaccgcg atgagaggag agatcacatg tgcctggtgg aagtggtcaa 31500 ggccgtcgat ctcgatacct atcagtaaaa cttgtttatt gcagcttata atggttacaa 31560 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 31620 tggtttgtcc aaactcatca atgtatctta actagttcta gacaattgac tctatgtggg 31680 atatgctcca gcgctacaac cttgaagtca ggcttcctgg atgtcagcat ctgactttgg 31740 ccagcacctg tcccgcggat ttgttccagt ccaactacag cgacccaccc taacagagat 31800 gaccaacaca accaacgcgg ccgccgctac cggacttaca tctaccacaa atacacccca 31860 agtttctgcc tttgtcaata actgggataa cttgggcatg tggtggttct ccatagcgct 31920 tatgtttgta tgccttatta ttatgtggct catctgctgc ctaaagcgca aacgcgcccg 31980 accacccatc tatagtccca tcattgtgct acacccaaac aatgatggaa tccatagatt 32040 ggacggactg aaacacatgt tcttttctct tacagtatga ttaaatgaga catgattcct 32100 cgagttttta tattactgac ccttgttgcg cttttttgtg cgtgctccac attggctgcg 32160 gtttctcaca tcgaagtaga ctgcattcca gccttcacag tctatttgct ttacggattt 32220 gtcaccctca cgctcatctg cagcctcatc actgtggtca tcgcctttat ccagtgcatt 32280 gactgggtct gtgtgcgctt tgcatatctc agacaccatc cccagtacag ggacaggact 32340 atagctgagc ttcttagaat tctttaatta tgaaatttac tgtgactttt ctgctgatta 32400 tttgcaccct atctgcgttt tgttccccga cctccaagcc tcaaagacat atatcatgca 32460 gattcactcg tatatggaat attccaagtt gctacaatga aaaaagcgat ctttccgaag 32520 cctggttata tgcaatcatc tctgttatgg tgttctgcag taccatctta gccctagcta 32580 tatatcccta ccttgacatt ggctggaacg caatagatgc catgaaccac ccaactttcc 32640 ccgcgcccgc tatgcttcca ctgcaacaag ttgttgccgg cggctttgtc ccagccaatc 32700 agcctcgccc accttctccc acccccactg aaatcagcta ctttaatcta acaggaggag 32760 atgactgaca ccctagatct agaaatggac ggaattatta cagagcagcg cctgctagaa 32820 agacgcaggg cagcggccga gcaacagcgc atgaatcaag agctccaaga catggttaac 32880 ttgcaccagt gcaaaagggg tatcttttgt ctggtaaagc aggccaaagt cacctacgac 32940 agtaatacca ccggacaccg ccttagctac aagttgccaa ccaagcgtca gaaattggtg 33000 gtcatggtgg gagaaaagcc cattaccata actcagcact cggtagaaac cgaaggctgc 33060 attcactcac cttgtcaagg acctgaggat ctctgcaccc ttattaagac cctgtgcggt 33120 ctcaaagatc ttattccctt taactaataa aaaaaaataa taaagcatca cttacttaaa 33180 atcagttagc aaatttctgt ccagtttatt cagcagcacc tccttgccct cctcccagct 33240 ctggtattgc agcttcctcc tggctgcaaa ctttctccac aatctaaatg gaatgtcagt 33300 ttcctcctgt tcctgtccat ccgcacccac tatcttcatg ttgttgcaga tgaagcgcgc 33360 aagaccgtct gaagatacct tcaaccccgt gtatccatat gacacggaaa ccggtcctcc 33420 aactgtgcct tttcttactc ctccctttgt atcccccaat gggtttcaag agagtccccc 33480 tggggtactc tctttgcgcc tatccgaacc tctagttacc tccaatggca tgcttgcgct 33540 caaaatgggc aacggcctct ctctggacga ggccggcaac cttacctccc aaaatgtaac 33600 cactgtgagc ccacctctca aaaaaaccaa gtcaaacata aacctggaaa tatctgcacc 33660 cctcacagtt acctcagaag ccctaactgt ggctgccgcc gcacctctaa tggtcgcggg 33720 caacacactc accatgcaat cacaggcccc gctaaccgtg cacgactcca aacttagcat 33780 tgccacccaa ggacccctca cagtgtcaga aggaaagcta gccctgcaaa catcaggccc 33840 cctcaccacc accgatagca gtacccttac tatcactgcc tcaccccctc taactactgc 33900 cactggtagc ttgggcattg acttgaaaga gcccatttat acacaaaatg gaaaactagg 33960 actaaagtac ggggctcctt tgcatgtaac agacgaccta aacactttga ccgtagcaac 34020 tggtccaggt gtgactatta ataatacttc cttgcaaact aaagttactg gagccttggg 34080 ttttgattca caaggcaata tgcaacttaa tgtagcagga ggactaagga ttgattctca 34140 aaacagacgc cttatacttg atgttagtta tccgtttgat gctcaaaacc aactaaatct 34200 aagactagga cagggccctc tttttataaa ctcagcccac aacttggata ttaactacaa 34260 caaaggcctt tacttgttta cagcttcaaa caattccaaa aagcttgagg ttaacctaag 34320 cactgccaag gggttgatgt ttgacgctac agccatagcc attaatgcag gagatgggct 34380 tgaatttggt tcacctaatg caccaaacac aaatcccctc aaaacaaaaa ttggccatgg 34440 cctagaattt gattcaaaca aggctatggt tcctaaacta ggaactggcc ttagttttga 34500 cagcacaggt gccattacag taggaaacaa aaataatgat aagctaaccc tatggacagg 34560 tccaaaacca gaagccaact gcataattga atacgggaaa caaaacccag atagcaaact 34620 aactttaatc cttgtaaaaa atggaggaat tgttaatgga tatgtaacgc taatgggagc 34680 ctcagactac gttaacacct tatttaaaaa caaaaatgtc tccattaatg tagaactata 34740 ctttgatgcc actggtcata tattaccaga ctcatcttct cttaaaacag atctagaact 34800 aaaatacaag caaaccgctg actttagtgc aagaggtttt atgccaagta ctacagcgta 34860 tccatttgtc cttcctaatg cgggaacaca taatgaaaat tatatttttg gtcaatgcta 34920 ctacaaagca agcgatggtg ccctttttcc gttggaagtt actgttatgc ttaataaacg 34980 cctgccagat agtcgcacat cctatgttat gactttttta tggtccttga atgctggtct 35040 agctccagaa actactcagg caaccctcat aacctcccca tttacctttt cctatattag 35100 agaagatgac taataaagaa tcgtttgtgg gccaccatgg ccttatgttt caacgtgttt 35160 atttttcaat tgcagaaaat ttcaagtcat ttttcattca gtagtatagc cccaccacca 35220 catagcttat acagatcacc gtaccttaat caaactcaca gaaccctagt attcaacctg 35280 ccacctccct cccaacacac agagtacaca gtcctttctc cccggctggc cttaaaaagc 35340 atcatatcat gggtaacaga catattctta ggtgttatat tccacacggt ttcctgtcga 35400 gccaaacgct catcagtgat attaataaac tccccgggca gctcacttaa gttcatgtcg 35460 ctgtccagct gctgagccac aggctgctgt ccaacttgcg gttgcttaac gggcggcgaa 35520 ggagaagtcc acgcctacat gggggtagag tcataatcgt gcatcaggat agggcggtgg 35580 tgctgcagca gcgcgcgaat aaactgctgc cgccgccgct ccgtcctgca ggaatacaac 35640 atggcagtgg tctcctcagc gatgattcgc accgcccgca gcataaggcg ccttgtcctc 35700 cgggcacagc agcgcaccct gatctcactt aaatcagcac agtaactgca gcacagcacc 35760 acaatattgt tcaaaatccc acagtgcaag gcgctgtatc caaagctcat ggcggggacc 35820 acagaaccca cgtggccatc ataccacaag cgcaggtaga ttaagtggcg acccctcata 35880 aacacgctgg acataaacat tacctctttt ggcatgttgt aattcaccac ctcccggtac 35940 catataaacc tctgattaaa catggcgcca tccaccacca tcctaaacca gctggccaaa 36000 acctgcccgc cggctataca ctgcagggaa ccgggactgg aacaatgaca gtggagagcc 36060 caggactcgt aaccatggat catcatgctc gtcatgatat caatgttggc acaacacagg 36120 cacacgtgca tacacttcct caggattaca agctcctccc gcgttagaac catatcccag 36180 ggaacaaccc attcctgaat cagcgtaaat cccacactgc agggaagacc tcgcacgtaa 36240 ctcacgttgt gcattgtcaa agtgttacat tcgggcagca gcggatgatc ctccagtatg 36300 gtagcgcggg tttctgtctc aaaaggaggt agacgatccc tactgtacgg agtgcgccga 36360 gacaaccgag atcgtgttgg tcgtagtgtc atgccaaatg gaacgccgga cgtagtcata 36420 tttcctgaag caaaaccagg tgcgggcgtg acaaacagat ctgcgtctcc ggtctcgccg 36480 cttagatcgc tctgtgtagt agttgtagta tatccactct ctcaaagcat ccaggcgccc 36540 cctggcttcg ggttctatgt aaactccttc atgcgccgct gccctgataa catccaccac 36600 cgcagaataa gccacaccca gccaacctac acattcgttc tgcgagtcac acacgggagg 36660 agcgggaaga gctggaagaa ccatgttttt ttttttattc caaaagatta tccaaaacct 36720 caaaatgaag atctattaag tgaacgcgct cccctccggt ggcgtggtca aactctacag 36780 ccaaagaaca gataatggca tttgtaagat gttgcacaat ggcttccaaa aggcaaacgg 36840 ccctcacgtc caagtggacg taaaggctaa acccttcagg gtgaatctcc tctataaaca 36900 ttccagcacc ttcaaccatg cccaaataat tctcatctcg ccaccttctc aatatatctc 36960 taagcaaatc ccgaatatta agtccggcca ttgtaaaaat ctgctccaga gcgccctcca 37020 ccttcagcct caagcagcga atcatgattg caaaaattca ggttcctcac agacctgtat 37080 aagattcaaa agcggaacat taacaaaaat accgcgatcc cgtaggtccc ttcgcagggc 37140 cagctgaaca taatcgtgca ggtctgcacg gaccagcgcg gccacttccc cgccaggaac 37200 catgacaaaa gaacccacac tgattatgac acgcatactc ggagctatgc taaccagcgt 37260 agccccgatg taagcttgtt gcatgggcgg cgatataaaa tgcaaggtgc tgctcaaaaa 37320 atcaggcaaa gcctcgcgca aaaaagaaag cacatcgtag tcatgctcat gcagataaag 37380 gcaggtaagc tccggaacca ccacagaaaa agacaccatt tttctctcaa acatgtctgc 37440 gggtttctgc ataaacacaa aataaaataa caaaaaaaca tttaaacatt agaagcctgt 37500 cttacaacag gaaaaacaac ccttataagc ataagacgga ctacggccat gccggcgtga 37560 ccgtaaaaaa actggtcacc gtgattaaaa agcaccaccg acagctcctc ggtcatgtcc 37620 ggagtcataa tgtaagactc ggtaaacaca tcaggttgat tcacatcggt cagtgctaaa 37680 aagcgaccga aatagcccgg gggaatacat acccgcaggc gtagagacaa cattacagcc 37740 cccataggag gtataacaaa attaatagga gagaaaaaca cataaacacc tgaaaaaccc 37800 tcctgcctag gcaaaatagc accctcccgc tccagaacaa catacagcgc ttccacagcg 37860 gcagccataa cagtcagcct taccagtaaa aaagaaaacc tattaaaaaa acaccactcg 37920 acacggcacc agctcaatca gtcacagtgt aaaaaagggc caagtgcaga gcgagtatat 37980 ataggactaa aaaatgacgt aacggttaaa gtccacaaaa aacacccaga aaaccgcacg 38040 cgaacctacg cccagaaacg aaagccaaaa aacccacaac ttcctcaaat cgtcacttcc 38100 gttttcccac gttacgtcac ttcccatttt aagaaaacta caattcccaa cacatacaag 38160 ttactccgcc ctaaaaccta cgtcacccgc cccgttccca cgccccgcgc cacgtcacaa 38220 actccacccc ctcattatca tattggcttc aatccaaaat aaggtatatt attgatgatg 38280 ttaattaatt taaatatcgc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 38340 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 38400 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 38460 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 38520 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 38580 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 38640 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 38700 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 38760 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 38820 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 38880 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 38940 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 39000 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 39060 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 39120 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 39180 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 39240 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 39300 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 39360 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 39420 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 39480 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 39540 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 39600 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 39660 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 39720 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 39780 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 39840 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 39900 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 39960 tggtatcttt atagtcctgt tgggtttcgc cacctctgac ttgagcgtcg atttttgtga 40020 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 40080 ctggcctttt gctggccttt tgctcacatg tacggggagt tctggctttc ggtgccagca 40140 tctgcgccac accaccgagc accatactgg caccgagaga aaacaggatg ccggtcatac 40200 caccggcccc aatggctgcc ccccatgctg caagggtggc tccggcggta aagaatgatc 40260 cggcaatggc ggcagccccc aggacaatct ggaatacgcc acctgacttg gccccggcga 40320 ctctgggaac aatatgaatt acagcgccat caggcagagt ctcatgtaac tgcgccgtta 40380 acccggacgt gctgacgtcc cgcccggcaa tccgtacctg ataccagccg tcgctcagtt 40440 tctgacgaaa cgccgggagc tgtgtggcca gtgcccggat ggcttcagcc cccgttttca 40500 cacgaaggtc gatgcggcga ccaaatcgtt gtaaatcccc gtaaaggcag atgcgcgcgc 40560 gccatgcccg gtgacgccag agggagtgtg tgcgtcgctg ccatttgtcg gtgtacctct 40620 ctcgtttgct cagttgttca ggaatatggt gcagcagctc gccgtcgccg cagtaaattg 40680 cggcgtgatt cggcactgat gaaccaaaac agcacagcag cacatcgccc ggctgtgccg 40740 ctgacaacgg cacctgatac agccccgtcg cctccagatt atccagatag agattctggc 40800 cgttacgcca ccagtcatcc tcacgatgaa agtccggcat ctcaatcccc gccagatgat 40860 aagcatcccg gaacagtgtg taacagtccg tcaccgaaaa ctcctccagt ctgctggcgt 40920 tatcctccgt cagctcgact ttttccagca gctccttgcc gagatgggat tcggttatct 40980 tgcctttgaa aaaatccagg taaccttccg catcatcgct cgcccgaccg acggcctcca 41040 cgaatgccga tttgccaacg gtgttcacac tgcggatata aaagtaataa tcatggcccg 41100 gtttgatatt gatactggcg gctatccagt acagcgccgt accaagataa cgcgtgctgg 41160 tttcaacctg tctgatatcc gcaatctgct tttccgagaa ccagaactca aactgtaccg 41220 tcgggtcata aacggcaaga tgcggcgtgg cggttatctg aaaatagccc ggcgtcagct 41280 caatcctcga cggtgctgcc ggtgcggcaa tccggaacga taccgacgcc ggatcgccct 41340 gctgccccca cgcatttacc gcccggactg tcagcctgta gttccccagc gccagttgcg 41400 tgaagcggta tgtggtttcc gtcgtccggg ccgtgctgac cagccgctca ctgccgtcgt 41460 ccgctgttac ggtcagacgg agcaggaaac tcacgccctt caccaccttc ggtgtgtccc 41520 atcgcgccag cacctgatat tccccgctgt ctgcagtgac ttctgcggtc aggtgctgca 41580 ccgctggcgg cgtgacacca ttcaccgtgc cactctgttc gccgtcaaag tgcgccccgt 41640 tatccacgat ggcctctttt tccggcacat gctgcacggc ggtgatggca tacgtgccgt 41700 cgtcgttctc acggatactc acgcagcgga acagtcgctg gcgcagcgtc ggcagcttca 41760 gctcccatac gctgtattca gcaacaccgt caggaacacg gctcactttt accttcacgc 41820 cgtcggtgac ggactgaacc tccacgctga ccggattgcc acttccgtca accaggctta 41880 tcagcgcggt accggaggat ggcagcgtga tttcacggtc gagcgtcagc gtccgggtct 41940 ggctgttcac cgccagcaca cgaccaccgg tgctgatacc ggcatagtca tcatcgcaga 42000 tttcaataac atcgcccggt acatggcgaa gcccttctgc gccgacgctg aaatccacgg 42060 tctgcgtttc cagcagttct gttttaatca gccacagccc ggcgcggtgt gcctgccccc 42120 ggctggtaca gccaaaggca tccatcttcg taacattacg accgtaacgg gcaatggcct 42180 gcgtatcttc aacaagctct gtcgccgtct cccagccgtt gttcgggtca atccagttca 42240 cctcaacggc attatggcgg tccttcaggg cgctgaagct gtagcggaac ggcgcggacc 42300 gctataaccg cactcgtcac cgcgataggt ccaggtgcag gtgttggcca gcatgatacg 42360 tcccggaaaa acagcgccat ccgtttccgt cggcgtggac agtacaaagg aggcactcac 42420 cgcgctcagt tcgctgcact gctcaatgcg ccagcggctg atcacctcct gctccggatc 42480 ggcgtaactg tttccgttga cgaagttcac cgcatccaga aaacgggcgt aaaccttacg 42540 ccggaccacc gttccgccga ccagactctg catatcttcc gccatcccgg tgaccatacc 42600 gtacaggtta gaaaccgtca gcgtggggcg cgtactggtg cctttgccat tcagttcaaa 42660 accgctcccc tgaatgggat acggctgata ctgtcgcccc tgccaggtga ccggctcacc 42720 tttttcgttc tgctcattac agaaaaaata acgttctcca ccgacctctg gtcaggtcga 42780 tttcccagag caccacgctg gccgactgct ccgcacgggt gcattcattc agtgtttcct 42840 gccggatatc ctgcatcagt tcaccacctg ttcaaactct gcgctgaact caacacgcag 42900 catactgacc cgcgacgacc attttgcgca ggtcaccttt atctgccgcc actcataagg 42960 cggcgtccac agaaaggatt tccagccccc gtgctcttcc agaaacgact ccagtaccgt 43020 ggcctcctca cgggggacag aaagcgtcac gctgtacgtt ttcaggttgg cattcagccc 43080 ggcaggcgct cgctgagaat agccatcacc aaagcgcacc tttcttacag aagggaccga 43140 agccacatcc ataccgggtt tcactttcca gcggaaggtc ttcatcgtcc acctccggag 43200 aacaggccac catcacgcat ctgtgtctga atttcatcac gggcaccctt gcgggccatg 43260 tcatacaccg ccttcagagc agccggacct atctgcccgt tcgtgccgtc gttgttaatc 43320 accacatggt tattctgctc aaacgtcccg gacgcctgcg accggctgtc tgccatgctg 43380 cccggtgtac cgacataacc gccggtggca tagccgcgca tcagccggta aagattcccc 43440 acgccaatcc ggctggttgc ctccttcgtg aagacaaact caccacggtg aacaatcccc 43500 gctggctcat atttgccgcc ggttcccgta aatcctccgg ttgcaaaatg gaatttcgcc 43560 gcagcggcct gaatggctgt accgcctgac gcggatgcgc cgccaccaac agccccgcca 43620 atggcgctgc cgatactccc gacaatcccc accattgcct gcttaagcag aatttctgtc 43680 atcatggaca gcacggaacg ggtgaagctg cgccagttct gctcactgcc ggtcagcatc 43740 gccgccatat tctgtgcaat accatcaaag gtctgcgtgg ctgcactttt tacctgcgac 43800 atactgtccg tggcgctctc ttcccactca ctccagccgg acttcaggcc tgccatccag 43860 ttcccgcgaa gctggtcttc agccgcccag gtctttttct gctctgacat gacgttattc 43920 agcgccagcg gattatcgcc atactgttcc ttcaggcgct gttccgtggc ttcccgttct 43980 gcctgccggt cagtcagccc ccggcttttc gcatcaatgg cggcccgttt tgcccgttgc 44040 tgctgtgcga atttatccgc ctgctgcgcc agcgcgttca ggcgctcctg atacgtaacc 44100 ttgtcgccaa gtgcagccag ctggcgtttg tactccagcg tctcatcttt atgcgccagc 44160 agggatttct cctgtgcaga cagctggcga cgttgcgccg cctcctccag taccgcgaac 44220 tgactctccg ccttccacaa atcccggcgc tgctggctga ttttctcatt tgctccggca 44280 tgcttctcca gcgtccggag ttctgcctga agcgtcagca gggcagcatg agcactgtct 44340 tcctgacgat cgcccgcaga caccttcacg ctggactgtt tcggcttttt cagcgtcgct 44400 tcataatcct ttttcgccgc cgccatcagc gtgttgtaat ccgcctgcag gattttcccg 44460 tctttcagtg ccttgttcag ttcttcctga cgggcggtat atttctccag cggcgtctgc 44520 agccgttcgt aagccttctg cgcctcttcg gtatatttca gccgtgacgc ttcggtatcg 44580 ctctgctgct gcgcattttt gtcctgttga gtctgctgct cagccttctt tcgggcggct 44640 tcaagcgcaa gacgggcctt ttcacgatca tcccagtaac gcgcccgcgc ttcatcgtta 44700 acaaaataat catccttgcg cagattccag atgtcgtctg ctttcttata cgcagcctct 44760 gccttaatca gcatctcctg cgcggtatca ggacgaccaa tatccagcac cgcatcccac 44820 atggatttga atgcccgcgc agtcctgtct gcccaggtct ccagcgtgcc catgttctct 44880 ttcaggcggc gggtctggtc atcaaaccct ttcgttgcgg cctcgttcgc cgcctgcaat 44940 gccccggctt catcgccgga acgctgcaac tgagcaacat acgcaatctg ctccgccgac 45000 acgttatgga actggcgagc catcgccgtc agccccgacg tcgggtctgt ggtcagcttc 45060 ccgaaggctt cagcgacctt tgtccacctc cacgccggat gcagaggaga aacgcgccac 45120 gatcttctag aggcctagaa ggccagagtg gcagagactg cattcgaaaa cgtttgaatt 45180 gataattatt atcatttgcg ggtcaattct tagaaaaact catcgagcat caaatgaaac 45240 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat 45300 gaaggagaaa actcaccgag gcagttccat aggattgcaa gatcctggta tcggtctgcg 45360 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta 45420 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc 45480 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca 45540 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg 45600 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca 45660 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg 45720 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc 45780 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg 45840 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat 45900 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa 45960 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg 46020 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgcgaaa 46080 acgtttgaat tgataattat tatcatttgc gggtcctttc cggcgatccg ccttgttacg 46140 gggcggcgac ctcgcgggtt ttcgctattt atgaaaattt tccggtttaa ggcgtttccg 46200 ttcttcttcg tcataactta atgtttttat ttaaaatacc ctctgaaaag aaaggaaacg 46260 acagctgaaa gcgagctttt tggcctctgt cgtttccttt ctctgttttt gtccgtggaa 46320 tgaacaatgg aagtcggcct cgtgatacgc ctatttttat aggttaatgt catgataata 46380 atggtttctt agcgatattt aaattaat 46408 <210> 9 <211> 45768 <212> DNA <213> Artificial Sequence <220> <223> recombinant vector sequence(SFTPC-GFP-miR142) <400> 9 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatatttgt ctagggagct 360 agcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tggctagcac 420 tagtattggc agatttgacc tcgtaaatac atagagatgg ctttgggaag gcactaggaa 480 agacagagaa aagagaagga gacagtcctc aaagctgatc gtatttgggt gagtattatt 540 ctcagggcaa atttaggatc agaggatgca gaaaggggag tctagagggg tagagtgtag 600 accacagggt gagtgagctg attcgaggat ggggagactg ggagcccacc agtgaccaga 660 gccagccctg ttcagggctg tccgggcaga agaaagcagt gtcagacctg gaatctgcca 720 tcagcacagc ctgcaattga cagacaagcc cagagcaaag aaggaagcac tgcacatgag 780 taagagcttg ccaccagtgg ggacagagtt tccagaatta ggaaaataat cactgggggc 840 aagtttgagg ttggtaccag atatgtggga ggaggcaagg taagggaaag agtacttgaa 900 gttggaactg gtccttgcag ggaaatgcac atttatgaaa ccccgaaaac tgatgtcaaa 960 gcacctcctg ccttgggcag agtcctctca gagtctacag gtgctgcctc cagaaccctc 1020 ttcctggagc gcatccctat gtatctagaa attctgctgg gaaatatgat ggtcagaccc 1080 ttggccacct gaaaggttca gggtggtaga agaaaaagga aagccacagg gcagcagggg 1140 caggtgccag caaggaaggc aggcacgcca ggaagacacc catggtgaga agtgcagatg 1200 gcccgagggc aagtttgctc aactcaccca ggtttgctct tgctggggcc aagaggactc 1260 atgtgccagg gccaagggcc cttgggggct ctcacagggg gcttatctgg gcttcggttc 1320 tggagggcca ggaacaaaca ggcttcaaag ccaagggctt ggctggcaca cagggggctt 1380 ggtccttcac ctctgtcccc tctccctacg gacacatata agaccctggt cacacctggg 1440 agaggaggag aggagagcat agcacctgca gcaagactag tacaccggga ctgaaaatga 1500 gacatattat ctgccacgga ggtgttatta ccgaagaaat ggccgccagt cttttggacc 1560 agctgatcga agaggtactg gctgataatc ttccacctcc tagccatttt gaaccaccta 1620 cccttcacga actgtatgat ttagacgtga cggcccccga agatcccaac gaggaggcgg 1680 tttcgcagat ttttcccgac tctgtaatgt tggcggtgca ggaagggatt gacttactca 1740 cttttccgcc ggcgcccggt tctccggagc cgcctcacct ttcccggcag cccgagcagc 1800 cggagcagag agccttgggt ccggtttcta tgccaaacct tgtaccggag gtgatcgatc 1860 ttacctgcca cgaggctggc tttccaccca gtgacgacga ggatgaagag ggtgaggagt 1920 ttgtgttaga ttatgtggag caccccgggc acggttgcag gtcttgtcat tatcaccgga 1980 ggaatacggg ggacccagat attatgtgtt cgctttgcta tatgaggacc tgtggcatgt 2040 ttgtctacag tcctgtgtct gaacctgagc ctgagcccga gccagaaccg gagcctgcaa 2100 gacctacccg ccgtcctaaa atggcgcctg ctatcctgag acgcccgaca tcacctgtgt 2160 ctagagaatg caatagtagt acggatagct gtgactccgg tccttctaac acacctcctg 2220 agatacaccc ggtggtcccg ctgtgcccca ttaaaccagt tgccgtgaga gttggtgggc 2280 gtcgccaggc tgtggaatgt atcgaggact tgcttaacga gcctgggcaa cctttggact 2340 tgagctgtaa acgccccagg ccataaggtg taaacctgtg acccctctcc ctcccccccc 2400 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt 2460 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt 2520 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt 2580 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct 2640 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt 2700 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt 2760 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa 2820 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta 2880 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa 2940 aacacgatga taagcttgcc acaacccggg atcctctaga gtcgacatgg aggcttggga 3000 gtgtttggaa gatttttctg ctgtgcgtaa cttgctggaa cagagctcta acagtacctc 3060 ttggttttgg aggtttctgt ggggctcatc ccaggcaaag ttagtctgca gaattaagga 3120 ggattacaag tgggaatttg aagagctttt gaaatcctgt ggtgagctgt ttgattcttt 3180 gaatctgggt caccaggcgc ttttccaaga gaaggtcatc aagactttgg atttttccac 3240 accggggcgc gctgcggctg ctgttgcttt tttgagtttt ataaaggata aatggagcga 3300 agaaacccat ctgagcgggg ggtacctgct ggattttctg gccatgcatc tgtggagagc 3360 ggttgtgaga cacaagaatc gcctgctact gttgtcttcc gtccgcccgg cgataatacc 3420 gacggaggag cagcagcagc agcaggagga agccaggcgg cggcggcagg agcagagccc 3480 atggaacccg agagccggcc tggaccctcg ggaatgaatg ttgtacaggt ggctgaactg 3540 tatccagaac tgagacgcat tttgacaatt acagaggatg ggcaggggct aaagggggta 3600 aagagggagc ggggggcttg tgaggctaca gaggaggcta ggaatctagc ttttagctta 3660 atgaccagac accgtcctga gtgtattact tttcaacaga tcaaggataa ttgcgctaat 3720 gagcttgatc tgctggcgca gaagtattcc atagagcagc tgaccactta ctggctgcag 3780 ccaggggatg attttgagga ggctattagg gtatatgcaa aggtggcact taggccagat 3840 tgcaagtaca agatcagcaa acttgtaaat atcaggaatt gttgctacat ttctgggaac 3900 ggggccgagg tggagataga tacggaggat agggtggcct ttagatgtag catgataaat 3960 atgtggccgg gggtgcttgg catggacggg gtggttatta tgaatgtaag gtttactggc 4020 cccaatttta gcggtacggt tttcctggcc aataccaacc ttatcctaca cggtgtaagc 4080 ttctatgggt ttaacaatac ctgtgtggaa gcctggaccg atgtaagggt tcggggctgt 4140 gccttttact gctgctggaa gggggtggtg tgtcgcccca aaagcagggc ttcaattaag 4200 aaatgcctct ttgaaaggtg taccttgggt atcctgtctg agggtaactc cagggtgcgc 4260 cacaatgtgg cctccgactg tggttgcttc atgctagtga aaagcgtggc tgtgattaag 4320 cataacatgg tatgtggcaa ctgcgaggac agggcctctc agatgctgac ctgctcggac 4380 ggcaactgtc acctgctgaa gaccattcac gtagccagcc actctcgcaa ggcctggcca 4440 gtgtttgagc ataacatact gacccgctgt tccttgcatt tgggtaacag gaggggggtg 4500 ttcctacctt accaatgcaa tttgagtcac actaagatat tgcttgagcc cgagagcatg 4560 tccaaggtga acctgaacgg ggtgtttgac atgaccatga agatctggaa ggtgctgagg 4620 tacgatgaga cccgcaccag gtgcagaccc tgcgagtgtg gcggtaaaca tattaggaac 4680 cagcctgtga tgctggatgt gaccgaggag ctgaggcccg atcacttggt gctggcctgc 4740 acccgcgctg agtttggctc tagcgatgaa gatacagatt gaagtactga aatgtgtggg 4800 cgtggcttaa gggtgggaaa gaatatataa ggtgggggtc ttatgtagtt ttgtatctgt 4860 tttgcagcag ccgccgccgc catgagcacc aactcgtttg atggaagcat tgtgagctca 4920 tatttgacaa cgcgcatgcc cccatgggcc ggggtgcgtc agaatgtgat gggctccagc 4980 attgatggtc gccccgtcct gcccgcaaac tctactacct tgacctacga gaccgtgtct 5040 ggaacgccgt tggagactgc agcctccgcc gccgcttcag ccgctgcagc caccgcccgc 5100 gggattgtga ctgactttgc tttcctgagc ccgcttgcaa gcagtgcagc ttcccgttca 5160 tccgcccgcg atgacaagtt gacggctctt ttggcacaat tggattcttt gacccgggaa 5220 cttaatgtcg tttctcagca gctgttggat ctgcgccagc aggtttctgc cctgaaggct 5280 tcctcccctc ccaatgcggt ttaacggtcc gggccagagt ggccaacata aataaaaaac 5340 cagactctgt ttggatttgg atcaagcaag tgtcttgctg tctttattta ggggttttgc 5400 gcgcgcggta ggcccgggac cagcggtctc ggtcgttgag ggtcctgtgt attttttcca 5460 ggacgtggta aaggtgactc tggatgttca gatacatggg cataagcccg tctctggggt 5520 ggaggtagca ccactgcaga gcttcatgct gcggggtggt gttgtagatg atccagtcgt 5580 agcaggagcg ctgggcgtgg tgcctaaaaa tgtctttcag tagcaagctg attgccaggg 5640 gcaggccctt ggtgtaagtg tttacaaagc ggttaagctg ggatgggtgc atacgtgggg 5700 atatgagatg catcttggac tgtattttta ggttggctat gttcccagcc atatccctcc 5760 ggggattcat gttgtgcaga accaccagca cagtgtatcc ggtgcacttg ggaaatttgt 5820 catgtagctt agaaggaaat gcgtggaaga acttggagac gcccttgtga cctccaagat 5880 tttccatgca ttcgtccata atgatggcaa tgggcccacg ggcggcggcc tgggcgaaga 5940 tatttctggg atcactaacg tcatagttgt gttccaggat gagatcgtca taggccattt 6000 ttacaaagcg cgggcggagg gtgccagact gcggtataat ggttccatcc ggcccagggg 6060 cgtagttacc ctcacagatt tgcatttccc acgctttgag ttcagatggg gggatcatgt 6120 ctacctgcgg ggcgatgaag aaaacggttt ccggggtagg ggagatcagc tgggaagaaa 6180 gcaggttcct gagcagctgc gacttaccgc agccggtggg cccgtaaatc acacctatta 6240 ccggctgcaa ctggtagtta agagagctgc agctgccgtc atccctgagc aggggggcca 6300 cttcgttaag catgtccctg actcgcatgt tttccctgac caaatccgcc agaaggcgct 6360 cgccgcccag cgatagcagt tcttgcaagg aagcaaagtt tttcaacggt ttgagaccgt 6420 ccgccgtagg catgcttttg agcgtttgac caagcagttc caggcggtcc cacagctcgg 6480 tcacctgctc tacggcatct cgatccagca tatctcctcg tttcgcgggt tggggcggct 6540 ttcgctgtac ggcagtagtc ggtgctcgtc cagacgggcc agggtcatgt ctttccacgg 6600 gcgcagggtc ctcgtcagcg tagtctgggt cacggtgaag gggtgcgctc cgggctgcgc 6660 gctggccagg gtgcgcttga ggctggtcct gctggtgctg aagcgctgcc ggtcttcgcc 6720 ctgcgcgtcg gccaggtagc atttgaccat ggtgtcatag tccagcccct ccgcggcgtg 6780 gcccttggcg cgcagcttgc ccttggagga ggcgccgcac gaggggcagt gcagactttt 6840 gagggcgtag agcttgggcg cgagaaatac cgattccggg gagtaggcat ccgcgccgca 6900 ggccccgcag acggtctcgc attccacgag ccaggtgagc tctggccgtt cggggtcaaa 6960 aaccaggttt cccccatgct ttttgatgcg tttcttacct ctggtttcca tgagccggtg 7020 tccacgctcg gtgacgaaaa ggctgtccgt gtccccgtat acagacttga gaggcctgtc 7080 ctcgagcggt gttccgcggt cctcctcgta tagaaactcg gaccactctg agacaaaggc 7140 tcgcgtccag gccagcacga aggaggctaa gtgggagggg tagcggtcgt tgtccactag 7200 ggggtccact cgctccaggg tgtgaagaca catgtcgccc tcttcggcat caaggaaggt 7260 gattggtttg taggtgtagg ccacgtgacc gggtgttcct gaaggggggc tataaaaggg 7320 ggtgggggcg cgttcgtcct cactctcttc cgcatcgctg tctgcgaggg ccagctgttg 7380 gggtgagtac tccctctgaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa 7440 aaacgaggag gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcatc 7500 catctggtca gaaaagacaa tctttttgtt gtcaagcttg gtggcaaacg acccgtagag 7560 ggcgttggac agcaacttgg cgatggagcg cagggtttgg tttttgtcgc gatcggcgcg 7620 ctccttggcc gcgatgttta gctgcacgta ttcgcgcgca acgcaccgcc attcgggaaa 7680 gacggtggtg cgctcgtcgg gcaccaggtg cacgcgccaa ccgcggttgt gcagggtgac 7740 aaggtcaacg ctggtggcta cctctccgcg taggcgctcg ttggtccagc agaggcggcc 7800 gcccttgcgc gagcagaatg gcggtagggg gtctagctgc gtctcgtccg gggggtctgc 7860 gtccacggta aagaccccgg gcagcaggcg cgcgtcgaag tagtctatct tgcatccttg 7920 caagtctagc gcctgctgcc atgcgcgggc ggcaagcgcg cgctcgtatg ggttgagtgg 7980 gggaccccat ggcatggggt gggtgagcgc ggaggcgtac atgccgcaaa tgtcgtaaac 8040 gtagaggggc tctctgagta ttccaagata tgtagggtag catcttccac cgcggatgct 8100 ggcgcgcacg taatcgtata gttcgtgcga gggagcgagg aggtcgggac cgaggttgct 8160 acgggcgggc tgctctgctc ggaagactat ctgcctgaag atggcatgtg agttggatga 8220 tatggttgga cgctggaaga cgttgaagct ggcgtctgtg agacctaccg cgtcacgcac 8280 gaaggaggcg taggagtcgc gcagcttgtt gaccagctcg gcggtgacct gcacgtctag 8340 ggcgcagtag tccagggttt ccttgatgat gtcatactta tcctgtccct tttttttcca 8400 cagctcgcgg ttgaggacaa actcttcgcg gtctttccag tactcttgga tcggaaaccc 8460 gtcggcctcc gaacggtaag agcctagcat gtagaactgg ttgacggcct ggtaggcgca 8520 gcatcccttt tctacgggta gcgcgtatgc ctgcgcggcc ttccggagcg aggtgtgggt 8580 gagcgcaaag gtgtccctga ccatgacttt gaggtactgg tatttgaagt cagtgtcgtc 8640 gcatccgccc tgctcccaga gcaaaaagtc cgtgcgcttt ttggaacgcg gatttggcag 8700 ggcgaaggtg acatcgttga agagtatctt tcccgcgcga ggcataaagt tgcgtgtgat 8760 gcggaagggt cccggcacct cggaacggtt gttaattacc tgggcggcga gcacgatctc 8820 gtcaaagccg ttgatgttgt ggcccacaat gtaaagttcc aagaagcgcg ggatgccctt 8880 gatggaaggc aattttttaa gttcctcgta ggtgagctct tcaggggagc tgagcccgtg 8940 ctctgaaagg gcccagtctg caagatgagg gttggaagcg acgaatgagc tccacaggtc 9000 acgggccatt agcatttgca ggtggtcgcg aaaggtccta aactggcgac ctatggccat 9060 tttttctggg gtgatgcagt agaaggtaag cgggtcttgt tcccagcggt cccatccaag 9120 gttcgcggct aggtctcgcg cggcagtcac tagaggctca tctccgccga acttcatgac 9180 cagcatgaag ggcacgagct gcttcccaaa ggcccccatc caagtatagg tctctacatc 9240 gtaggtgaca aagagacgct cggtgcgagg atgcgagccg atcgggaaga actggatctc 9300 ccgccaccaa ttggaggagt ggctattgat gtggtgaaag tagaagtccc tgcgacgggc 9360 cgaacactcg tgctggcttt tgtaaaaacg tgcgcagtac tggcagcggt gcacgggctg 9420 tacatcctgc acgaggttga cctgacgacc gcgcacaagg aagcagagtg ggaatttgag 9480 cccctcgcct ggcgggtttg gctggtggtc ttctacttcg gctgcttgtc cttgaccgtc 9540 tggctgctcg aggggagtta cggtggatcg gaccaccacg ccgcgcgagc ccaaagtcca 9600 gatgtccgcg cgcggcggtc ggagcttgat gacaacatcg cgcagatggg agctgtccat 9660 ggtctggagc tcccgcggcg tcaggtcagg cgggagctcc tgcaggttta cctcgcatag 9720 acgggtcagg gcgcgggcta gatccaggtg atacctaatt tccaggggct ggttggtggc 9780 ggcgtcgatg gcttgcaaga ggccgcatcc ccgcggcgcg actacggtac cgcgcggcgg 9840 gcggtgggcc gcgggggtgt ccttggatga tgcatctaaa agcggtgacg cgggcgagcc 9900 cccggaggta gggggggctc cggacccgcc gggagagggg gcaggggcac gtcggcgccg 9960 cgcgcgggca ggagctggtg ctgcgcgcgt aggttgctgg cgaacgcgac gacgcggcgg 10020 ttgatctcct gaatctggcg cctctgcgtg aagacgacgg gcccggtgag cttgaacctg 10080 aaagagagtt cgacagaatc aatttcggtg tcgttgacgg cggcctggcg caaaatctcc 10140 tgcacgtctc ctgagttgtc ttgataggcg atctcggcca tgaactgctc gatctcttcc 10200 tcctggagat ctccgcgtcc ggctcgctcc acggtggcgg cgaggtcgtt ggaaatgcgg 10260 gccatgagct gcgagaaggc gttgaggcct ccctcgttcc agacgcggct gtagaccacg 10320 cccccttcgg catcgcgggc gcgcatgacc acctgcgcga gattgagctc cacgtgccgg 10380 gcgaagacgg cgtagtttcg caggcgctga aagaggtagt tgagggtggt ggcggtgtgt 10440 tctgccacga agaagtacat aacccagcgt cgcaacgtgg attcgttgat atcccccaag 10500 gcctcaaggc gctccatggc ctcgtagaag tccacggcga agttgaaaaa ctgggagttg 10560 cgcgccgaca cggttaactc ctcctccaga agacggatga gctcggcgac agtgtcgcgc 10620 acctcgcgct caaaggctac aggggcctct tcttcttctt caatctcctc ttccataagg 10680 gcctcccctt cttcttcttc tggcggcggt gggggagggg ggacacggcg gcgacgacgg 10740 cgcaccggga ggcggtcgac aaagcgctcg atcatctccc cgcggcgacg gcgcatggtc 10800 tcggtgacgg cgcggccgtt ctcgcggggg cgcagttgga agacgccgcc cgtcatgtcc 10860 cggttatggg ttggcggggg gctgccatgc ggcagggata cggcgctaac gatgcatctc 10920 aacaattgtt gtgtaggtac tccgccgccg agggacctga gcgagtccgc atcgaccgga 10980 tcggaaaacc tctcgagaaa ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc 11040 gtggcgggcg gcagcgggcg gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg 11100 taattaaagt aggcggtctt gagacggcgg atggtcgaca gaagcaccat gtccttgggt 11160 ccggcctgct gaatgcgcag gcggtcggcc atgccccagg cttcgttttg acatcggcgc 11220 aggtctttgt agtagtcttg catgagcctt tctaccggca cttcttcttc tccttcctct 11280 tgtcctgcat ctcttgcatc tatcgctgcg gcggcggcgg agtttggccg taggtggcgc 11340 cctcttcctc ccatgcgtgt gaccccgaag cccctcatcg gctgaagcag ggctaggtcg 11400 gcgacaacgc gctcggctaa tatggcctgc tgcacctgcg tgagggtaga ctggaagtca 11460 tccatgtcca caaagcggtg gtatgcgccc gtgttgatgg tgtaagtgca gttggccata 11520 acggaccagt taacggtctg gtgacccggc tgcgagagct cggtgtacct gagacgcgag 11580 taagccctcg agtcaaatac gtagtcgttg caagtccgca ccaggtactg gtatcccacc 11640 aaaaagtgcg gcggcggctg gcggtagagg ggccagcgta gggtggccgg ggctccgggg 11700 gcgagatctt ccaacataag gcgatgatat ccgtagatgt acctggacat ccaggtgatg 11760 ccggcggcgg tggtggaggc gcgcggaaag tcgcggacgc ggttccagat gttgcgcagc 11820 ggcaaaaagt gctccatggt cgggacgctc tggccggtca ggcgcgcgca atcgttgacg 11880 ctctagaccg tgcaaaagga gagcctgtaa gcgggcactc ttccgtggtc tggtggataa 11940 attcgcaagg gtatcatggc ggacgaccgg ggttcgagcc ccgtatccgg ccgtccgccg 12000 tgatccatgc ggttaccgcc cgcgtgtcga acccaggtgt gcgacgtcag acaacggggg 12060 agtgctcctt ttggcttcct tccaggcgcg gcggctgctg cgctagcttt tttggccact 12120 ggccgcgcgc agcgtaagcg gttaggctgg aaagcgaaag cattaagtgg ctcgctccct 12180 gtagccggag ggttattttc caagggttga gtcgcgggac ccccggttcg agtctcggac 12240 cggccggact gcggcgaacg ggggtttgcc tccccgtcat gcaagacccc gcttgcaaat 12300 tcctccggaa acagggacga gccccttttt tgcttttccc agatgcatcc ggtgctgcgg 12360 cagatgcgcc cccctcctca gcagcggcaa gagcaagagc agcggcagac atgcagggca 12420 ccctcccctc ctcctaccgc gtcaggaggg gcgacatccg cggttgacgc ggcagcagat 12480 ggtgattacg aacccccgcg gcgccgggcc cggcactacc tggacttgga ggagggcgag 12540 ggcctggcgc ggctaggagc gccctctcct gagcggcacc caagggtgca gctgaagcgt 12600 gatacgcgtg aggcgtacgt gccgcggcag aacctgtttc gcgaccgcga gggagaggag 12660 cccgaggaga tgcgggatcg aaagttccac gcagggcgcg agctgcggca tggcctgaat 12720 cgcgagcggt tgctgcgcga ggaggacttt gagcccgacg cgcgaaccgg gattagtccc 12780 gcgcgcgcac acgtggcggc cgccgacctg gtaaccgcat acgagcagac ggtgaaccag 12840 gagattaact ttcaaaaaag ctttaacaac cacgtgcgta cgcttgtggc gcgcgaggag 12900 gtggctatag gactgatgca tctgtgggac tttgtaagcg cgctggagca aaacccaaat 12960 agcaagccgc tcatggcgca gctgttcctt atagtgcagc acagcaggga caacgaggca 13020 ttcagggatg cgctgctaaa catagtagag cccgagggcc gctggctgct cgatttgata 13080 aacatcctgc agagcatagt ggtgcaggag cgcagcttga gcctggctga caaggtggcc 13140 gccatcaact attccatgct tagcctgggc aagttttacg cccgcaagat ataccatacc 13200 ccttacgttc ccatagacaa ggaggtaaag atcgaggggt tctacatgcg catggcgctg 13260 aaggtgctta ccttgagcga cgacctgggc gtttatcgca acgagcgcat ccacaaggcc 13320 gtgagcgtga gccggcggcg cgagctcagc gaccgcgagc tgatgcacag cctgcaaagg 13380 gccctggctg gcacgggcag cggcgataga gaggccgagt cctactttga cgcgggcgct 13440 gacctgcgct gggccccaag ccgacgcgcc ctggaggcag ctggggccgg acctgggctg 13500 gcggtggcac ccgcgcgcgc tggcaacgtc ggcggcgtgg aggaatatga cgaggacgat 13560 gagtacgagc cagaggacgg cgagtactaa gcggtgatgt ttctgatcag atgatgcaag 13620 acgcaacgga cccggcggtg cgggcggcgc tgcagagcca gccgtccggc cttaactcca 13680 cggacgactg gcgccaggtc atggaccgca tcatgtcgct gactgcgcgc aatcctgacg 13740 cgttccggca gcagccgcag gccaaccggc tctccgcaat tctggaagcg gtggtcccgg 13800 cgcgcgcaaa ccccacgcac gagaaggtgc tggcgatcgt aaacgcgctg gccgaaaaca 13860 gggccatccg gcccgacgag gccggcctgg tctacgacgc gctgcttcag cgcgtggctc 13920 gttacaacag cggcaacgtg cagaccaacc tggaccggct ggtgggggat gtgcgcgagg 13980 ccgtggcgca gcgtgagcgc gcgcagcagc agggcaacct gggctccatg gttgcactaa 14040 acgccttcct gagtacacag cccgccaacg tgccgcgggg acaggaggac tacaccaact 14100 ttgtgagcgc actgcggcta atggtgactg agacaccgca aagtgaggtg taccagtctg 14160 ggccagacta ttttttccag accagtagac aaggcctgca gaccgtaaac ctgagccagg 14220 ctttcaaaaa cttgcagggg ctgtgggggg tgcgggctcc cacaggcgac cgcgcgaccg 14280 tgtctagctt gctgacgccc aactcgcgcc tgttgctgct gctaatagcg cccttcacgg 14340 acagtggcag cgtgtcccgg gacacatacc taggtcactt gctgacactg taccgcgagg 14400 ccataggtca ggcgcatgtg gacgagcata ctttccagga gattacaagt gtcagccgcg 14460 cgctggggca ggaggacacg ggcagcctgg aggcaaccct aaactacctg ctgaccaacc 14520 ggcggcagaa gatcccctcg ttgcacagtt taaacagcga ggaggagcgc attttgcgct 14580 acgtgcagca gagcgtgagc cttaacctga tgcgcgacgg ggtaacgccc agcgtggcgc 14640 tggacatgac cgcgcgcaac atggaaccgg gcatgtatgc ctcaaaccgg ccgtttatca 14700 accgcctaat ggactacttg catcgcgcgg ccgccgtgaa ccccgagtat ttcaccaatg 14760 ccatcttgaa cccgcactgg ctaccgcccc ctggtttcta caccggggga ttcgaggtgc 14820 ccgagggtaa cgatggattc ctctgggacg acatagacga cagcgtgttt tccccgcaac 14880 cgcagaccct gctagagttg caacagcgcg agcaggcaga ggcggcgctg cgaaaggaaa 14940 gcttccgcag gccaagcagc ttgtccgatc taggcgctgc ggccccgcgg tcagatgcta 15000 gtagcccatt tccaagcttg atagggtctc ttaccagcac tcgcaccacc cgcccgcgcc 15060 tgctgggcga ggaggagtac ctaaacaact cgctgctgca gccgcagcgc gaaaaaaacc 15120 tgcctccggc atttcccaac aacgggatag agagcctagt ggacaagatg agtagatgga 15180 agacgtacgc gcaggagcac agggacgtgc caggcccgcg cccgcccacc cgtcgtcaaa 15240 ggcacgaccg tcagcggggt ctggtgtggg aggacgatga ctcggcagac gacagcagcg 15300 tcctggattt gggagggagt ggcaacccgt ttgcgcacct tcgccccagg ctggggagaa 15360 tgttttaaaa aaaaaaaaaa gcatgatgca aaataaaaaa ctcaccaagg ccatggcacc 15420 gagcgttggt tttcttgtat tccccttagt atgcggcgcg cggcgatgta tgaggaaggt 15480 cctcctccct cctacgagag tgtggtgagc gcggcgccag tggcggcggc gctgggttct 15540 cccttcgatg ctcccctgga cccgccgttt gtgcctccgc ggtacctgcg gcctaccggg 15600 gggagaaaca gcatccgtta ctctgagttg gcacccctat tcgacaccac ccgtgtgtac 15660 ctggtggaca acaagtcaac ggatgtggca tccctgaact accagaacga ccacagcaac 15720 tttctgacca cggtcattca aaacaatgac tacagcccgg gggaggcaag cacacagacc 15780 atcaatcttg acgaccggtc gcactggggc ggcgacctga aaaccatcct gcataccaac 15840 atgccaaatg tgaacgagtt catgtttacc aataagttta aggcgcgggt gatggtgtcg 15900 cgcttgccta ctaaggacaa tcaggtggag ctgaaatacg agtgggtgga gttcacgctg 15960 cccgagggca actactccga gaccatgacc atagacctta tgaacaacgc gatcgtggag 16020 cactacttga aagtgggcag acagaacggg gttctggaaa gcgacatcgg ggtaaagttt 16080 gacacccgca acttcagact ggggtttgac cccgtcactg gtcttgtcat gcctggggta 16140 tatacaaacg aagccttcca tccagacatc attttgctgc caggatgcgg ggtggacttc 16200 acccacagcc gcctgagcaa cttgttgggc atccgcaagc ggcaaccctt ccaggagggc 16260 tttaggatca cctacgatga tctggagggt ggtaacattc ccgcactgtt ggatgtggac 16320 gcctaccagg cgagcttgaa agatgacacc gaacagggcg ggggtggcgc aggcggcagc 16380 aacagcagtg gcagcggcgc ggaagagaac tccaacgcgg cagccgcggc aatgcagccg 16440 gtggaggaca tgaacgatca tgccattcgc ggcgacacct ttgccacacg ggctgaggag 16500 aagcgcgctg aggccgaagc agcggccgaa gctgccgccc ccgctgcgca acccgaggtc 16560 gagaagcctc agaagaaacc ggtgatcaaa cccctgacag aggacagcaa gaaacgcagt 16620 tacaacctaa taagcaatga cagcaccttc acccagtacc gcagctggta ccttgcatac 16680 aactacggcg accctcagac cggaatccgc tcatggaccc tgctttgcac tcctgacgta 16740 acctgcggct cggagcaggt ctactggtcg ttgccagaca tgatgcaaga ccccgtgacc 16800 ttccgctcca cgcgccagat cagcaacttt ccggtggtgg gcgccgagct gttgcccgtg 16860 cactccaaga gcttctacaa cgaccaggcc gtctactccc aactcatccg ccagtttacc 16920 tctctgaccc acgtgttcaa tcgctttccc gagaaccaga ttttggcgcg cccgccagcc 16980 cccaccatca ccaccgtcag tgaaaacgtt cctgctctca cagatcacgg gacgctaccg 17040 ctgcgcaaca gcatcggagg agtccagcga gtgaccatta ctgacgccag acgccgcacc 17100 tgcccctacg tttacaaggc cctgggcata gtctcgccgc gcgtcctatc gagccgcact 17160 ttttgagcaa gcatgtccat ccttatatcg cccagcaata acacaggctg gggcctgcgc 17220 ttcccaagca agatgtttgg cggggccaag aagcgctccg accaacaccc agtgcgcgtg 17280 cgcgggcact accgcgcgcc ctggggcgcg cacaaacgcg gccgcactgg gcgcaccacc 17340 gtcgatgacg ccatcgacgc ggtggtggag gaggcgcgca actacacgcc cacgccgcca 17400 ccagtgtcca cagtggacgc ggccattcag accgtggtgc gcggagcccg gcgctatgct 17460 aaaatgaaga gacggcggag gcgcgtagca cgtcgccacc gccgccgacc cggcactgcc 17520 gcccaacgcg cggcggcggc cctgcttaac cgcgcacgtc gcaccggccg acgggcggcc 17580 atgcgagcag ctcgaaggct ggccgcgggt attgtcactg tgccccccag gtccaggcga 17640 cgagcggccg ccgcagcagc cgcggccatt agtgctatga ctcagggtcg caggggcaac 17700 gtgtattggg tgcgcgactc ggttagcggc ctgcgcgtgc ccgtgcgcac ccgccccccg 17760 cgcaactaga ttgcaagaaa aaactactta gactcgtact gttgtatgta tccagcggcg 17820 gcggcgcgca acgaagctat gtccaagcgc aaaatcaaag aagagatgct ccaggtcatc 17880 gcgccggaga tctatggccc cccgaagaag gaagagcagg attacaagcc ccgaaagcta 17940 aagcgggtca aaaagaaaaa gaaagatgat gatgatgaac ttgacgacga ggtggaactg 18000 ctgcacgcta ccgcgcccag gcgacgggta cagtggaaag gtcgacgcgt aaaacgtgtt 18060 ttgcgacccg gcaccaccgt agtctttacg cccggtgagc gctccacccg cacctacaag 18120 cgcgtgtatg atgaggtgta cggcgacgag gacctgcttg agcaggccaa cgagcgcctc 18180 ggggagtttg cctacggaaa gcggcataag gacatgctgg cgttgccgct ggacgagggc 18240 aacccaacac ctagcctaaa gcccgtaaca ctgcagcagg tgctgcccgc gcttgcaccg 18300 tccgaagaaa agcgcggcct aaagcgcgag tctggtgact tggcacccac cgtgcagctg 18360 atggtaccca agcgccagcg actggaagat gtcttggaaa aaatgaccgt ggaacctggg 18420 ctggagcccg aggtccgcgt gcggccaatc aagcaggtgg cgccgggact gggcgtgcag 18480 accgtggacg ttcagatacc cactaccagt agcaccagta ttgccaccgc cacagagggc 18540 atggagacac aaacgtcccc ggttgcctca gcggtggcgg atgccgcggt gcaggcggtc 18600 gctgcggccg cgtccaagac ctctacggag gtgcaaacgg acccgtggat gtttcgcgtt 18660 tcagcccccc ggcgcccgcg ccgttcgagg aagtacggcg ccgccagcgc gctactgccc 18720 gaatatgccc tacatccttc cattgcgcct acccccggct atcgtggcta cacctaccgc 18780 cccagaagac gagcaactac ccgacgccga accaccactg gaacccgccg ccgccgtcgc 18840 cgtcgccagc ccgtgctggc cccgatttcc gtgcgcaggg tggctcgcga aggaggcagg 18900 accctggtgc tgccaacagc gcgctaccac cccagcatcg tttaaaagcc ggtctttgtg 18960 gttcttgcag atatggccct cacctgccgc ctccgtttcc cggtgccggg attccgagga 19020 agaatgcacc gtaggagggg catggccggc cacggcctga cgggcggcat gcgtcgtgcg 19080 caccaccggc ggcggcgcgc gtcgcaccgt cgcatgcgcg gcggtatcct gcccctcctt 19140 attccactga tcgccgcggc gattggcgcc gtgcccggaa ttgcatccgt ggccttgcag 19200 gcgcagagac actgattaaa aacaagttgc atgtggaaaa atcaaaataa aaagtctgga 19260 ctctcacgct cgcttggtcc tgtaactatt ttgtagaatg gaagacatca actttgcgtc 19320 tctggccccg cgacacggct cgcgcccgtt catgggaaac tggcaagata tcggcaccag 19380 caatatgagc ggtggcgcct tcagctgggg ctcgctgtgg agcggcatta aaaatttcgg 19440 ttccaccgtt aagaactatg gcagcaaggc ctggaacagc agcacaggcc agatgctgag 19500 ggataagttg aaagagcaaa atttccaaca aaaggtggta gatggcctgg cctctggcat 19560 tagcggggtg gtggacctgg ccaaccaggc agtgcaaaat aagattaaca gtaagcttga 19620 tccccgccct cccgtagagg agcctccacc ggccgtggag acagtgtctc cagaggggcg 19680 tggcgaaaag cgtccgcgcc ccgacaggga agaaactctg gtgacgcaaa tagacgagcc 19740 tccctcgtac gaggaggcac taaagcaagg cctgcccacc acccgtccca tcgcgcccat 19800 ggctaccgga gtgctgggcc agcacacacc cgtaacgctg gacctgcctc cccccgccga 19860 cacccagcag aaacctgtgc tgccaggccc gaccgccgtt gttgtaaccc gtcctagccg 19920 cgcgtccctg cgccgcgccg ccagcggtcc gcgatcgttg cggcccgtag ccagtggcaa 19980 ctggcaaagc acactgaaca gcatcgtggg tctgggggtg caatccctga agcgccgacg 20040 atgcttctga tagctaacgt gtcgtatgtg tgtcatgtat gcgtccatgt cgccgccaga 20100 ggagctgctg agccgccgcg cgcccgcttt ccaagatggc taccccttcg atgatgccgc 20160 agtggtctta catgcacatc tcgggccagg acgcctcgga gtacctgagc cccgggctgg 20220 tgcagtttgc ccgcgccacc gagacgtact tcagcctgaa taacaagttt agaaacccca 20280 cggtggcgcc tacgcacgac gtgaccacag accggtccca gcgtttgacg ctgcggttca 20340 tccctgtgga ccgtgaggat actgcgtact cgtacaaggc gcggttcacc ctagctgtgg 20400 gtgataaccg tgtgctggac atggcttcca cgtactttga catccgcggc gtgctggaca 20460 ggggccctac ttttaagccc tactctggca ctgcctacaa cgccctggct cccaagggtg 20520 ccccaaatcc ttgcgaatgg gatgaagctg ctactgctct tgaaataaac ctagaagaag 20580 aggacgatga caacgaagac gaagtagacg agcaagctga gcagcaaaaa actcacgtat 20640 ttgggcaggc gccttattct ggtataaata ttacaaagga gggtattcaa ataggtgtcg 20700 aaggtcaaac acctaaatat gccgataaaa catttcaacc tgaacctcaa ataggagaat 20760 ctcagtggta cgaaacagaa attaatcatg cagctgggag agtcctaaaa aagactaccc 20820 caatgaaacc atgttacggt tcatatgcaa aacccacaaa tgaaaatgga gggcaaggca 20880 ttcttgtaaa gcaacaaaat ggaaagctag aaagtcaagt ggaaatgcaa tttttctcaa 20940 ctactgaggc agccgcaggc aatggtgata acttgactcc taaagtggta ttgtacagtg 21000 aagatgtaga tatagaaacc ccagacactc atatttctta catgcccact attaaggaag 21060 gtaactcacg agaactaatg ggccaacaat ctatgcccaa caggcctaat tacattgctt 21120 ttagggacaa ttttattggt ctaatgtatt acaacagcac gggtaatatg ggtgttctgg 21180 cgggccaagc atcgcagttg aatgctgttg tagatttgca agacagaaac acagagcttt 21240 cataccagct tttgcttgat tccattggtg atagaaccag gtacttttct atgtggaatc 21300 aggctgttga cagctatgat ccagatgtta gaattattga aaatcatgga actgaagatg 21360 aacttccaaa ttactgcttt ccactgggag gtgtgattaa tacagagact cttaccaagg 21420 taaaacctaa aacaggtcag gaaaatggat gggaaaaaga tgctacagaa ttttcagata 21480 aaaatgaaat aagagttgga aataattttg ccatggaaat caatctaaat gccaacctgt 21540 ggagaaattt cctgtactcc aacatagcgc tgtatttgcc cgacaagcta aagtacagtc 21600 cttccaacgt aaaaatttct gataacccaa acacctacga ctacatgaac aagcgagtgg 21660 tggctcccgg gctagtggac tgctacatta accttggagc acgctggtcc cttgactata 21720 tggacaacgt caacccattt aaccaccacc gcaatgctgg cctgcgctac cgctcaatgt 21780 tgctgggcaa tggtcgctat gtgcccttcc acatccaggt gcctcagaag ttctttgcca 21840 ttaaaaacct ccttctcctg ccgggctcat acacctacga gtggaacttc aggaaggatg 21900 ttaacatggt tctgcagagc tccctaggaa atgacctaag ggttgacgga gccagcatta 21960 agtttgatag catttgcctt tacgccacct tcttccccat ggcccacaac accgcctcca 22020 cgcttgaggc catgcttaga aacgacacca acgaccagtc ctttaacgac tatctctccg 22080 ccgccaacat gctctaccct atacccgcca acgctaccaa cgtgcccata tccatcccct 22140 cccgcaactg ggcggctttc cgcggctggg ccttcacgcg ccttaagact aaggaaaccc 22200 catcactggg ctcgggctac gacccttatt acacctactc tggctctata ccctacctag 22260 atggaacctt ttacctcaac cacaccttta agaaggtggc cattaccttt gactcttctg 22320 tcagctggcc tggcaatgac cgcctgctta cccccaacga gtttgaaatt aagcgctcag 22380 ttgacgggga gggttacaac gttgcccagt gtaacatgac caaagactgg ttcctggtac 22440 aaatgctagc taactataac attggctacc agggcttcta tatcccagag agctacaagg 22500 accgcatgta ctccttcttt agaaacttcc agcccatgag ccgtcaggtg gtggatgata 22560 ctaaatacaa ggactaccaa caggtgggca tcctacacca acacaacaac tctggatttg 22620 ttggctacct tgcccccacc atgcgcgaag gacaggccta ccctgctaac ttcccctatc 22680 cgcttatagg caagaccgca gttgacagca ttacccagaa aaagtttctt tgcgatcgca 22740 ccctttggcg catcccattc tccagtaact ttatgtccat gggcgcactc acagacctgg 22800 gccaaaacct tctctacgcc aactccgccc acgcgctaga catgactttt gaggtggatc 22860 ccatggacga gcccaccctt ctttatgttt tgtttgaagt ctttgacgtg gtccgtgtgc 22920 accagccgca ccgcggcgtc atcgaaaccg tgtacctgcg cacgcccttc tcggccggca 22980 acgccacaac ataaagaagc aagcaacatc aacaacagct gccgccatgg gctccagtga 23040 gcaggaactg aaagccattg tcaaagatct tggttgtggg ccatattttt tgggcaccta 23100 tgacaagcgc tttccaggct ttgtttctcc acacaagctc gcctgcgcca tagtcaatac 23160 ggccggtcgc gagactgggg gcgtacactg gatggccttt gcctggaacc cgcactcaaa 23220 aacatgctac ctctttgagc cctttggctt ttctgaccag cgactcaagc aggtttacca 23280 gtttgagtac gagtcactcc tgcgccgtag cgccattgct tcttcccccg accgctgtat 23340 aacgctggaa aagtccaccc aaagcgtaca ggggcccaac tcggccgcct gtggactatt 23400 ctgctgcatg tttctccacg cctttgccaa ctggccccaa actcccatgg atcacaaccc 23460 caccatgaac cttattaccg gggtacccaa ctccatgctc aacagtcccc aggtacagcc 23520 caccctgcgt cgcaaccagg aacagctcta cagcttcctg gagcgccact cgccctactt 23580 ccgcagccac agtgcgcaga ttaggagcgc cacttctttt tgtcacttga aaaacatgta 23640 aaaataatgt actagagaca ctttcaataa aggcaaatgc ttttatttgt acactctcgg 23700 gtgattattt acccccaccc ttgccgtctg cgccgtttaa aaatcaaagg ggttctgccg 23760 cgcatcgcta tgcgccactg gcagggacac gttgcgatac tggtgtttag tgctccactt 23820 aaactcaggc acaaccatcc gcggcagctc ggtgaagttt tcactccaca ggctgcgcac 23880 catcaccaac gcgtttagca ggtcgggcgc cgatatcttg aagtcgcagt tggggcctcc 23940 gccctgcgcg cgcgagttgc gatacacagg gttgcagcac tggaacacta tcagcgccgg 24000 gtggtgcacg ctggccagca cgctcttgtc ggagatcaga tccgcgtcca ggtcctccgc 24060 gttgctcagg gcgaacggag tcaactttgg tagctgcctt cccaaaaagg gcgcgtgccc 24120 aggctttgag ttgcactcgc accgtagtgg catcaaaagg tgaccgtgcc cggtctgggc 24180 gttaggatac agcgcctgca taaaagcctt gatctgctta aaagccacct gagcctttgc 24240 gccttcagag aagaacatgc cgcaagactt gccggaaaac tgattggccg gacacgcggc 24300 gtcgtgcacg cagcaccttg cgtcggtgtt ggagatctgc accacatttc ggccccaccg 24360 gttcttcacg atcttggcct tgctagactg ctccttcagc gcgcgctgcc cgttttcgct 24420 cgtcacatcc atttcaatca cgtgctcctt atttatcata atgcttccgt gtagacactt 24480 aagctcgcct tcgatctcag cgcagcggtg cagccacaac gcgcagcccg tgggctcgtg 24540 atgcttgtag gtcacctctg caaacgactg caggtacgcc tgcaggaatc gccccatcat 24600 cgtcacaaag gtcttgttgc tggtgaaggt cagctgcaac ccgcggtgct cctcgttcag 24660 ccaggtcttg catacggccg ccagagcttc cacttggtca ggcagtagtt tgaagttcgc 24720 ctttagatcg ttatccacgt ggtacttgtc catcagcgcg cgcgcagcct ccatgccctt 24780 ctcccacgca gacacgatcg gcacactcag cgggttcatc accgtaattt cactttccgc 24840 ttcgctgggc tcttcctctt cctcttgcgt ccgcatacca cgcgccactg ggtcgtcttc 24900 attcagccgc cgcactgtgc gcttacctcc tttgccatgc ttgattagca ccggtgggtt 24960 gctgaaaccc accatttgta gcgccacatc ttctctttct tcctcgctgt ccacgattac 25020 ctctggtgat ggcgggcgct cgggcttggg agaagggcgc ttctttttct tcttgggcgc 25080 aatggccaaa tccgccgccg aggtcgatgg ccgcgggctg ggtgtgcgcg gcaccagcgc 25140 gtcttgtgat gagtcttcct cgtcctcgga ctcgatacgc cgcctcatcc gcttttttgg 25200 gggcgcccgg ggaggcggcg gcgacgggga cggggacgac acgtcctcca tggttggggg 25260 acgtcgcgcc gcaccgcgtc cgcgctcggg ggtggtttcg cgctgctcct cttcccgact 25320 ggccatttcc ttctcctata ggcagaaaaa gatcatggag tcagtcgaga agaaggacag 25380 cctaaccgcc ccctctgagt tcgccaccac cgcctccacc gatgccgcca acgcgcctac 25440 caccttcccc gtcgaggcac ccccgcttga ggaggaggaa gtgattatcg agcaggaccc 25500 aggttttgta agcgaagacg acgaggaccg ctcagtacca acagaggata aaaagcaaga 25560 ccaggacaac gcagaggcaa acgaggaaca agtcgggcgg ggggacgaaa ggcatggcga 25620 ctacctagat gtgggagacg acgtgctgtt gaagcatctg cagcgccagt gcgccattat 25680 ctgcgacgcg ttgcaagagc gcagcgatgt gcccctcgcc atagcggatg tcagccttgc 25740 ctacgaacgc cacctattct caccgcgcgt accccccaaa cgccaagaaa acggcacatg 25800 cgagcccaac ccgcgcctca acttctaccc cgtatttgcc gtgccagagg tgcttgccac 25860 ctatcacatc tttttccaaa actgcaagat acccctatcc tgccgtgcca accgcagccg 25920 agcggacaag cagctggcct tgcggcaggg cgctgtcata cctgatatcg cctcgctcaa 25980 cgaagtgcca aaaatctttg agggtcttgg acgcgacgag aagcgcgcgg caaacgctct 26040 gcaacaggaa aacagcgaaa atgaaagtca ctctggagtg ttggtggaac tcgagggtga 26100 caacgcgcgc ctagccgtac taaaacgcag catcgaggtc acccactttg cctacccggc 26160 acttaaccta ccccccaagg tcatgagcac agtcatgagt gagctgatcg tgcgccgtgc 26220 gcagcccctg gagagggatg caaatttgca agaacaaaca gaggagggcc tacccgcagt 26280 tggcgacgag cagctagcgc gctggcttca aacgcgcgag cctgccgact tggaggagcg 26340 acgcaaacta atgatggccg cagtgctcgt taccgtggag cttgagtgca tgcagcggtt 26400 ctttgctgac ccggagatgc agcgcaagct agaggaaaca ttgcactaca cctttcgaca 26460 gggctacgta cgccaggcct gcaagatctc caacgtggag ctctgcaacc tggtctccta 26520 ccttggaatt ttgcacgaaa accgccttgg gcaaaacgtg cttcattcca cgctcaaggg 26580 cgaggcgcgc cgcgactacg tccgcgactg cgtttactta tttctatgct acacctggca 26640 gacggccatg ggcgtttggc agcagtgctt ggaggagtgc aacctcaagg agctgcagaa 26700 actgctaaag caaaacttga aggacctatg gacggccttc aacgagcgct ccgtggccgc 26760 gcacctggcg gacatcattt tccccgaacg cctgcttaaa accctgcaac agggtctgcc 26820 agacttcacc agtcaaagca tgttgcagaa ctttaggaac tttatcctag agcgctcagg 26880 aatcttgccc gccacctgct gtgcacttcc tagcgacttt gtgcccatta agtaccgcga 26940 atgccctccg ccgctttggg gccactgcta ccttctgcag ctagccaact accttgccta 27000 ccactctgac ataatggaag acgtgagcgg tgacggtcta ctggagtgtc actgtcgctg 27060 caacctatgc accccgcacc gctccctggt ttgcaattcg cagctgctta acgaaagtca 27120 aattatcggt acctttgagc tgcagggtcc ctcgcctgac gaaaagtccg cggctccggg 27180 gttgaaactc actccggggc tgtggacgtc ggcttacctt cgcaaatttg tacctgagga 27240 ctaccacgcc cacgagatta ggttctacga agaccaatcc cgcccgccta atgcggagct 27300 taccgcctgc gtcattaccc agggccacat tcttggccaa ttgcaagcca tcaacaaagc 27360 ccgccaagag tttctgctac gaaagggacg gggggtttac ttggaccccc agtccggcga 27420 ggagctcaac ccaatccccc cgccgccgca gccctatcag cagcagccgc gggcccttgc 27480 ttcccaggat ggcacccaaa aagaagctgc agctgccgcc gccacccacg gacgaggagg 27540 aatactggga cagtcaggca gaggaggttt tggacgagga ggaggaggac atgatggaag 27600 actgggagag cctagacgag gaagcttccg aggtcgaaga ggtgtcagac gaaacaccgt 27660 caccctcggt cgcattcccc tcgccggcgc cccagaaatc ggcaaccggt tccagcatgg 27720 ctacaacctc cgctcctcag gcgccgccgg cactgcccgt tcgccgaccc aaccgtagat 27780 gggacaccac tggaaccagg gccggtaagt ccaagcagcc gccgccgtta gcccaagagc 27840 aacaacagcg ccaaggctac cgctcatggc gcgggcacaa gaacgccata gttgcttgct 27900 tgcaagactg tgggggcaac atctccttcg cccgccgctt tcttctctac catcacggcg 27960 tggccttccc ccgtaacatc ctgcattact accgtcatct ctacagccca tactgcaccg 28020 gcggcagcgg cagcaacagc agcggccaca cagaagcaaa ggcgaccgga tagcaagact 28080 ctgacaaagc ccaagaaatc cacagcggcg gcagcagcag gaggaggagc gctgcgtctg 28140 gcgcccaacg aacccgtatc gacccgcgag cttagaaaca ggatttttcc cactctgtat 28200 gctatatttc aacagagcag gggccaagaa caagagctga aaataaaaaa caggtctctg 28260 cgatccctca cccgcagctg cctgtatcac aaaagcgaag atcagcttcg gcgcacgctg 28320 gaagacgcgg aggctctctt cagtaaatac tgcgcgctga ctcttaagga ctagtttcgc 28380 gccctttctc aaatttaagc gcgaaaacta cgtcatctcc agcggccaca cccggcgcca 28440 gcacctgttg tcagcgccat tatgagcaag gaaattccca cgccctacat gtggagttac 28500 cagccacaaa tgggacttgc ggctggagct gcccaagact actcaacccg aataaactac 28560 atgagcgcgg gaccccacat gatatcccgg gtcaacggaa tacgcgccca ccgaaaccga 28620 attctcctgg aacaggcggc tattaccacc acacctcgta ataaccttaa tccccgtagt 28680 tggcccgctg ccctggtgta ccaggaaagt cccgctccca ccactgtggt acttcccaga 28740 gacgcccagg ccgaagttca gatgactaac tcaggggcgc agcttgcggg cggctttcgt 28800 cacagggtgc ggtcgcccgg gcagggtata actcacctga caatcagagg gcgaggtatt 28860 cagctcaacg acgagtcggt gagctcctcg cttggtctcc gtccggacgg gacatttcag 28920 atcggcggcg ccggccgctc ttcattcacg cctcgtcagg caatcctaac tctgcagacc 28980 tcgtcctctg agccgcgctc tggaggcatt ggaactctgc aatttattga ggagtttgtg 29040 ccatcggtct actttaaccc cttctcggga cctcccggcc actatccgga tcaatttatt 29100 cctaactttg acgcggtaaa ggactcggcg gacggctacg actgaggcca atgaggccat 29160 gttaagtgga gaggcagagc aactgcgcct gaaacacctg gtccactgtc gccgccacaa 29220 gtgctttgcc cgcgactccg gtgagttttg ctactttgaa ttgcccgagg atcatatcga 29280 gggcccggcg cacggcgtcc ggcttaccgc ccagggagag cttgcccgta gcctgattcg 29340 ggagtttacc cagcgccccc tgctagttga gcgggacagg ggaccctgtg ttctcactgt 29400 gatttgcaac tgtcctaacc ctggattaca tcaagatctc ctaggctagc actagtcgtt 29460 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 29520 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 29580 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 29640 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 29700 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 29760 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 29820 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 29880 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 29940 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat 30000 ccacgctgtt ttgacctcca tagaagacac cggcccagaa gcttgctagc tccataaagt 30060 aggaaacact acatcactcc ataaagtagg aaacactaca gatatctcca taaagtagga 30120 aacactacat cactccataa agtaggaaac actacaaagc ttgtagcggc cgccaccatg 30180 actgccctga ccgaaggtgc taagctgttt gagaaggaga ttccgtacat caccgagctg 30240 gaaggggacg tcgaaggaat gaagttcatc atcaagggag aaggaaccgg ggacgctacg 30300 actggaacca ttaaggccaa gtatatctgt accactggag atctgccagt gccttgggcc 30360 acccttgtgt caaccctctc gtatggagtg cagtgttttg ctaagtaccc tagccacatt 30420 aaggacttct tcaaatccgc catgccggaa ggttataccc aagagcgcac catttctttt 30480 gagggagatg gagtgtacaa gacccgcgcg atggtcacct atgagagggg atctatctac 30540 aaccgggtga ctctgactgg agaaaacttt aagaaggacg ggcatattct tcggaagaat 30600 gtcgccttcc agtgccctcc cagcatcctt tacattctcc ccgacactgt gaacaacgga 30660 atccgcgtgg agttcaatca agcctacgac atcgaggggg tgacggagaa gctggtgacc 30720 aagtgtagcc agatgaatcg gccactggcc ggttcagcgg ctgtccacat tccgcgctac 30780 catcatatca cttatcacac taagctctcc aaagaccgcg atgagaggag agatcacatg 30840 tgcctggtgg aagtggtcaa ggccgtcgat ctcgatacct atcagtaaaa cttgtttatt 30900 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 30960 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta actagttcta 31020 gacaattgac tctatgtggg atatgctcca gcgctacaac cttgaagtca ggcttcctgg 31080 atgtcagcat ctgactttgg ccagcacctg tcccgcggat ttgttccagt ccaactacag 31140 cgacccaccc taacagagat gaccaacaca accaacgcgg ccgccgctac cggacttaca 31200 tctaccacaa atacacccca agtttctgcc tttgtcaata actgggataa cttgggcatg 31260 tggtggttct ccatagcgct tatgtttgta tgccttatta ttatgtggct catctgctgc 31320 ctaaagcgca aacgcgcccg accacccatc tatagtccca tcattgtgct acacccaaac 31380 aatgatggaa tccatagatt ggacggactg aaacacatgt tcttttctct tacagtatga 31440 ttaaatgaga catgattcct cgagttttta tattactgac ccttgttgcg cttttttgtg 31500 cgtgctccac attggctgcg gtttctcaca tcgaagtaga ctgcattcca gccttcacag 31560 tctatttgct ttacggattt gtcaccctca cgctcatctg cagcctcatc actgtggtca 31620 tcgcctttat ccagtgcatt gactgggtct gtgtgcgctt tgcatatctc agacaccatc 31680 cccagtacag ggacaggact atagctgagc ttcttagaat tctttaatta tgaaatttac 31740 tgtgactttt ctgctgatta tttgcaccct atctgcgttt tgttccccga cctccaagcc 31800 tcaaagacat atatcatgca gattcactcg tatatggaat attccaagtt gctacaatga 31860 aaaaagcgat ctttccgaag cctggttata tgcaatcatc tctgttatgg tgttctgcag 31920 taccatctta gccctagcta tatatcccta ccttgacatt ggctggaacg caatagatgc 31980 catgaaccac ccaactttcc ccgcgcccgc tatgcttcca ctgcaacaag ttgttgccgg 32040 cggctttgtc ccagccaatc agcctcgccc accttctccc acccccactg aaatcagcta 32100 ctttaatcta acaggaggag atgactgaca ccctagatct agaaatggac ggaattatta 32160 cagagcagcg cctgctagaa agacgcaggg cagcggccga gcaacagcgc atgaatcaag 32220 agctccaaga catggttaac ttgcaccagt gcaaaagggg tatcttttgt ctggtaaagc 32280 aggccaaagt cacctacgac agtaatacca ccggacaccg ccttagctac aagttgccaa 32340 ccaagcgtca gaaattggtg gtcatggtgg gagaaaagcc cattaccata actcagcact 32400 cggtagaaac cgaaggctgc attcactcac cttgtcaagg acctgaggat ctctgcaccc 32460 ttattaagac cctgtgcggt ctcaaagatc ttattccctt taactaataa aaaaaaataa 32520 taaagcatca cttacttaaa atcagttagc aaatttctgt ccagtttatt cagcagcacc 32580 tccttgccct cctcccagct ctggtattgc agcttcctcc tggctgcaaa ctttctccac 32640 aatctaaatg gaatgtcagt ttcctcctgt tcctgtccat ccgcacccac tatcttcatg 32700 ttgttgcaga tgaagcgcgc aagaccgtct gaagatacct tcaaccccgt gtatccatat 32760 gacacggaaa ccggtcctcc aactgtgcct tttcttactc ctccctttgt atcccccaat 32820 gggtttcaag agagtccccc tggggtactc tctttgcgcc tatccgaacc tctagttacc 32880 tccaatggca tgcttgcgct caaaatgggc aacggcctct ctctggacga ggccggcaac 32940 cttacctccc aaaatgtaac cactgtgagc ccacctctca aaaaaaccaa gtcaaacata 33000 aacctggaaa tatctgcacc cctcacagtt acctcagaag ccctaactgt ggctgccgcc 33060 gcacctctaa tggtcgcggg caacacactc accatgcaat cacaggcccc gctaaccgtg 33120 cacgactcca aacttagcat tgccacccaa ggacccctca cagtgtcaga aggaaagcta 33180 gccctgcaaa catcaggccc cctcaccacc accgatagca gtacccttac tatcactgcc 33240 tcaccccctc taactactgc cactggtagc ttgggcattg acttgaaaga gcccatttat 33300 acacaaaatg gaaaactagg actaaagtac ggggctcctt tgcatgtaac agacgaccta 33360 aacactttga ccgtagcaac tggtccaggt gtgactatta ataatacttc cttgcaaact 33420 aaagttactg gagccttggg ttttgattca caaggcaata tgcaacttaa tgtagcagga 33480 ggactaagga ttgattctca aaacagacgc cttatacttg atgttagtta tccgtttgat 33540 gctcaaaacc aactaaatct aagactagga cagggccctc tttttataaa ctcagcccac 33600 aacttggata ttaactacaa caaaggcctt tacttgttta cagcttcaaa caattccaaa 33660 aagcttgagg ttaacctaag cactgccaag gggttgatgt ttgacgctac agccatagcc 33720 attaatgcag gagatgggct tgaatttggt tcacctaatg caccaaacac aaatcccctc 33780 aaaacaaaaa ttggccatgg cctagaattt gattcaaaca aggctatggt tcctaaacta 33840 ggaactggcc ttagttttga cagcacaggt gccattacag taggaaacaa aaataatgat 33900 aagctaaccc tatggacagg tccaaaacca gaagccaact gcataattga atacgggaaa 33960 caaaacccag atagcaaact aactttaatc cttgtaaaaa atggaggaat tgttaatgga 34020 tatgtaacgc taatgggagc ctcagactac gttaacacct tatttaaaaa caaaaatgtc 34080 tccattaatg tagaactata ctttgatgcc actggtcata tattaccaga ctcatcttct 34140 cttaaaacag atctagaact aaaatacaag caaaccgctg actttagtgc aagaggtttt 34200 atgccaagta ctacagcgta tccatttgtc cttcctaatg cgggaacaca taatgaaaat 34260 tatatttttg gtcaatgcta ctacaaagca agcgatggtg ccctttttcc gttggaagtt 34320 actgttatgc ttaataaacg cctgccagat agtcgcacat cctatgttat gactttttta 34380 tggtccttga atgctggtct agctccagaa actactcagg caaccctcat aacctcccca 34440 tttacctttt cctatattag agaagatgac taataaagaa tcgtttgtgg gccaccatgg 34500 ccttatgttt caacgtgttt atttttcaat tgcagaaaat ttcaagtcat ttttcattca 34560 gtagtatagc cccaccacca catagcttat acagatcacc gtaccttaat caaactcaca 34620 gaaccctagt attcaacctg ccacctccct cccaacacac agagtacaca gtcctttctc 34680 cccggctggc cttaaaaagc atcatatcat gggtaacaga catattctta ggtgttatat 34740 tccacacggt ttcctgtcga gccaaacgct catcagtgat attaataaac tccccgggca 34800 gctcacttaa gttcatgtcg ctgtccagct gctgagccac aggctgctgt ccaacttgcg 34860 gttgcttaac gggcggcgaa ggagaagtcc acgcctacat gggggtagag tcataatcgt 34920 gcatcaggat agggcggtgg tgctgcagca gcgcgcgaat aaactgctgc cgccgccgct 34980 ccgtcctgca ggaatacaac atggcagtgg tctcctcagc gatgattcgc accgcccgca 35040 gcataaggcg ccttgtcctc cgggcacagc agcgcaccct gatctcactt aaatcagcac 35100 agtaactgca gcacagcacc acaatattgt tcaaaatccc acagtgcaag gcgctgtatc 35160 caaagctcat ggcggggacc acagaaccca cgtggccatc ataccacaag cgcaggtaga 35220 ttaagtggcg acccctcata aacacgctgg acataaacat tacctctttt ggcatgttgt 35280 aattcaccac ctcccggtac catataaacc tctgattaaa catggcgcca tccaccacca 35340 tcctaaacca gctggccaaa acctgcccgc cggctataca ctgcagggaa ccgggactgg 35400 aacaatgaca gtggagagcc caggactcgt aaccatggat catcatgctc gtcatgatat 35460 caatgttggc acaacacagg cacacgtgca tacacttcct caggattaca agctcctccc 35520 gcgttagaac catatcccag ggaacaaccc attcctgaat cagcgtaaat cccacactgc 35580 agggaagacc tcgcacgtaa ctcacgttgt gcattgtcaa agtgttacat tcgggcagca 35640 gcggatgatc ctccagtatg gtagcgcggg tttctgtctc aaaaggaggt agacgatccc 35700 tactgtacgg agtgcgccga gacaaccgag atcgtgttgg tcgtagtgtc atgccaaatg 35760 gaacgccgga cgtagtcata tttcctgaag caaaaccagg tgcgggcgtg acaaacagat 35820 ctgcgtctcc ggtctcgccg cttagatcgc tctgtgtagt agttgtagta tatccactct 35880 ctcaaagcat ccaggcgccc cctggcttcg ggttctatgt aaactccttc atgcgccgct 35940 gccctgataa catccaccac cgcagaataa gccacaccca gccaacctac acattcgttc 36000 tgcgagtcac acacgggagg agcgggaaga gctggaagaa ccatgttttt ttttttattc 36060 caaaagatta tccaaaacct caaaatgaag atctattaag tgaacgcgct cccctccggt 36120 ggcgtggtca aactctacag ccaaagaaca gataatggca tttgtaagat gttgcacaat 36180 ggcttccaaa aggcaaacgg ccctcacgtc caagtggacg taaaggctaa acccttcagg 36240 gtgaatctcc tctataaaca ttccagcacc ttcaaccatg cccaaataat tctcatctcg 36300 ccaccttctc aatatatctc taagcaaatc ccgaatatta agtccggcca ttgtaaaaat 36360 ctgctccaga gcgccctcca ccttcagcct caagcagcga atcatgattg caaaaattca 36420 ggttcctcac agacctgtat aagattcaaa agcggaacat taacaaaaat accgcgatcc 36480 cgtaggtccc ttcgcagggc cagctgaaca taatcgtgca ggtctgcacg gaccagcgcg 36540 gccacttccc cgccaggaac catgacaaaa gaacccacac tgattatgac acgcatactc 36600 ggagctatgc taaccagcgt agccccgatg taagcttgtt gcatgggcgg cgatataaaa 36660 tgcaaggtgc tgctcaaaaa atcaggcaaa gcctcgcgca aaaaagaaag cacatcgtag 36720 tcatgctcat gcagataaag gcaggtaagc tccggaacca ccacagaaaa agacaccatt 36780 tttctctcaa acatgtctgc gggtttctgc ataaacacaa aataaaataa caaaaaaaca 36840 tttaaacatt agaagcctgt cttacaacag gaaaaacaac ccttataagc ataagacgga 36900 ctacggccat gccggcgtga ccgtaaaaaa actggtcacc gtgattaaaa agcaccaccg 36960 acagctcctc ggtcatgtcc ggagtcataa tgtaagactc ggtaaacaca tcaggttgat 37020 tcacatcggt cagtgctaaa aagcgaccga aatagcccgg gggaatacat acccgcaggc 37080 gtagagacaa cattacagcc cccataggag gtataacaaa attaatagga gagaaaaaca 37140 cataaacacc tgaaaaaccc tcctgcctag gcaaaatagc accctcccgc tccagaacaa 37200 catacagcgc ttccacagcg gcagccataa cagtcagcct taccagtaaa aaagaaaacc 37260 tattaaaaaa acaccactcg acacggcacc agctcaatca gtcacagtgt aaaaaagggc 37320 caagtgcaga gcgagtatat ataggactaa aaaatgacgt aacggttaaa gtccacaaaa 37380 aacacccaga aaaccgcacg cgaacctacg cccagaaacg aaagccaaaa aacccacaac 37440 ttcctcaaat cgtcacttcc gttttcccac gttacgtcac ttcccatttt aagaaaacta 37500 caattcccaa cacatacaag ttactccgcc ctaaaaccta cgtcacccgc cccgttccca 37560 cgccccgcgc cacgtcacaa actccacccc ctcattatca tattggcttc aatccaaaat 37620 aaggtatatt attgatgatg ttaattaatt taaatatcgc aggtggcact tttcggggaa 37680 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 37740 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 37800 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 37860 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 37920 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 37980 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 38040 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 38100 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 38160 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 38220 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 38280 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 38340 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 38400 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 38460 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 38520 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 38580 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 38640 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 38700 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 38760 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 38820 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 38880 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 38940 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 39000 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 39060 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 39120 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 39180 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 39240 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 39300 agcttccagg gggaaacgcc tggtatcttt atagtcctgt tgggtttcgc cacctctgac 39360 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 39420 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tacggggagt 39480 tctggctttc ggtgccagca tctgcgccac accaccgagc accatactgg caccgagaga 39540 aaacaggatg ccggtcatac caccggcccc aatggctgcc ccccatgctg caagggtggc 39600 tccggcggta aagaatgatc cggcaatggc ggcagccccc aggacaatct ggaatacgcc 39660 acctgacttg gccccggcga ctctgggaac aatatgaatt acagcgccat caggcagagt 39720 ctcatgtaac tgcgccgtta acccggacgt gctgacgtcc cgcccggcaa tccgtacctg 39780 ataccagccg tcgctcagtt tctgacgaaa cgccgggagc tgtgtggcca gtgcccggat 39840 ggcttcagcc cccgttttca cacgaaggtc gatgcggcga ccaaatcgtt gtaaatcccc 39900 gtaaaggcag atgcgcgcgc gccatgcccg gtgacgccag agggagtgtg tgcgtcgctg 39960 ccatttgtcg gtgtacctct ctcgtttgct cagttgttca ggaatatggt gcagcagctc 40020 gccgtcgccg cagtaaattg cggcgtgatt cggcactgat gaaccaaaac agcacagcag 40080 cacatcgccc ggctgtgccg ctgacaacgg cacctgatac agccccgtcg cctccagatt 40140 atccagatag agattctggc cgttacgcca ccagtcatcc tcacgatgaa agtccggcat 40200 ctcaatcccc gccagatgat aagcatcccg gaacagtgtg taacagtccg tcaccgaaaa 40260 ctcctccagt ctgctggcgt tatcctccgt cagctcgact ttttccagca gctccttgcc 40320 gagatgggat tcggttatct tgcctttgaa aaaatccagg taaccttccg catcatcgct 40380 cgcccgaccg acggcctcca cgaatgccga tttgccaacg gtgttcacac tgcggatata 40440 aaagtaataa tcatggcccg gtttgatatt gatactggcg gctatccagt acagcgccgt 40500 accaagataa cgcgtgctgg tttcaacctg tctgatatcc gcaatctgct tttccgagaa 40560 ccagaactca aactgtaccg tcgggtcata aacggcaaga tgcggcgtgg cggttatctg 40620 aaaatagccc ggcgtcagct caatcctcga cggtgctgcc ggtgcggcaa tccggaacga 40680 taccgacgcc ggatcgccct gctgccccca cgcatttacc gcccggactg tcagcctgta 40740 gttccccagc gccagttgcg tgaagcggta tgtggtttcc gtcgtccggg ccgtgctgac 40800 cagccgctca ctgccgtcgt ccgctgttac ggtcagacgg agcaggaaac tcacgccctt 40860 caccaccttc ggtgtgtccc atcgcgccag cacctgatat tccccgctgt ctgcagtgac 40920 ttctgcggtc aggtgctgca ccgctggcgg cgtgacacca ttcaccgtgc cactctgttc 40980 gccgtcaaag tgcgccccgt tatccacgat ggcctctttt tccggcacat gctgcacggc 41040 ggtgatggca tacgtgccgt cgtcgttctc acggatactc acgcagcgga acagtcgctg 41100 gcgcagcgtc ggcagcttca gctcccatac gctgtattca gcaacaccgt caggaacacg 41160 gctcactttt accttcacgc cgtcggtgac ggactgaacc tccacgctga ccggattgcc 41220 acttccgtca accaggctta tcagcgcggt accggaggat ggcagcgtga tttcacggtc 41280 gagcgtcagc gtccgggtct ggctgttcac cgccagcaca cgaccaccgg tgctgatacc 41340 ggcatagtca tcatcgcaga tttcaataac atcgcccggt acatggcgaa gcccttctgc 41400 gccgacgctg aaatccacgg tctgcgtttc cagcagttct gttttaatca gccacagccc 41460 ggcgcggtgt gcctgccccc ggctggtaca gccaaaggca tccatcttcg taacattacg 41520 accgtaacgg gcaatggcct gcgtatcttc aacaagctct gtcgccgtct cccagccgtt 41580 gttcgggtca atccagttca cctcaacggc attatggcgg tccttcaggg cgctgaagct 41640 gtagcggaac ggcgcggacc gctataaccg cactcgtcac cgcgataggt ccaggtgcag 41700 gtgttggcca gcatgatacg tcccggaaaa acagcgccat ccgtttccgt cggcgtggac 41760 agtacaaagg aggcactcac cgcgctcagt tcgctgcact gctcaatgcg ccagcggctg 41820 atcacctcct gctccggatc ggcgtaactg tttccgttga cgaagttcac cgcatccaga 41880 aaacgggcgt aaaccttacg ccggaccacc gttccgccga ccagactctg catatcttcc 41940 gccatcccgg tgaccatacc gtacaggtta gaaaccgtca gcgtggggcg cgtactggtg 42000 cctttgccat tcagttcaaa accgctcccc tgaatgggat acggctgata ctgtcgcccc 42060 tgccaggtga ccggctcacc tttttcgttc tgctcattac agaaaaaata acgttctcca 42120 ccgacctctg gtcaggtcga tttcccagag caccacgctg gccgactgct ccgcacgggt 42180 gcattcattc agtgtttcct gccggatatc ctgcatcagt tcaccacctg ttcaaactct 42240 gcgctgaact caacacgcag catactgacc cgcgacgacc attttgcgca ggtcaccttt 42300 atctgccgcc actcataagg cggcgtccac agaaaggatt tccagccccc gtgctcttcc 42360 agaaacgact ccagtaccgt ggcctcctca cgggggacag aaagcgtcac gctgtacgtt 42420 ttcaggttgg cattcagccc ggcaggcgct cgctgagaat agccatcacc aaagcgcacc 42480 tttcttacag aagggaccga agccacatcc ataccgggtt tcactttcca gcggaaggtc 42540 ttcatcgtcc acctccggag aacaggccac catcacgcat ctgtgtctga atttcatcac 42600 gggcaccctt gcgggccatg tcatacaccg ccttcagagc agccggacct atctgcccgt 42660 tcgtgccgtc gttgttaatc accacatggt tattctgctc aaacgtcccg gacgcctgcg 42720 accggctgtc tgccatgctg cccggtgtac cgacataacc gccggtggca tagccgcgca 42780 tcagccggta aagattcccc acgccaatcc ggctggttgc ctccttcgtg aagacaaact 42840 caccacggtg aacaatcccc gctggctcat atttgccgcc ggttcccgta aatcctccgg 42900 ttgcaaaatg gaatttcgcc gcagcggcct gaatggctgt accgcctgac gcggatgcgc 42960 cgccaccaac agccccgcca atggcgctgc cgatactccc gacaatcccc accattgcct 43020 gcttaagcag aatttctgtc atcatggaca gcacggaacg ggtgaagctg cgccagttct 43080 gctcactgcc ggtcagcatc gccgccatat tctgtgcaat accatcaaag gtctgcgtgg 43140 ctgcactttt tacctgcgac atactgtccg tggcgctctc ttcccactca ctccagccgg 43200 acttcaggcc tgccatccag ttcccgcgaa gctggtcttc agccgcccag gtctttttct 43260 gctctgacat gacgttattc agcgccagcg gattatcgcc atactgttcc ttcaggcgct 43320 gttccgtggc ttcccgttct gcctgccggt cagtcagccc ccggcttttc gcatcaatgg 43380 cggcccgttt tgcccgttgc tgctgtgcga atttatccgc ctgctgcgcc agcgcgttca 43440 ggcgctcctg atacgtaacc ttgtcgccaa gtgcagccag ctggcgtttg tactccagcg 43500 tctcatcttt atgcgccagc agggatttct cctgtgcaga cagctggcga cgttgcgccg 43560 cctcctccag taccgcgaac tgactctccg ccttccacaa atcccggcgc tgctggctga 43620 ttttctcatt tgctccggca tgcttctcca gcgtccggag ttctgcctga agcgtcagca 43680 gggcagcatg agcactgtct tcctgacgat cgcccgcaga caccttcacg ctggactgtt 43740 tcggcttttt cagcgtcgct tcataatcct ttttcgccgc cgccatcagc gtgttgtaat 43800 ccgcctgcag gattttcccg tctttcagtg ccttgttcag ttcttcctga cgggcggtat 43860 atttctccag cggcgtctgc agccgttcgt aagccttctg cgcctcttcg gtatatttca 43920 gccgtgacgc ttcggtatcg ctctgctgct gcgcattttt gtcctgttga gtctgctgct 43980 cagccttctt tcgggcggct tcaagcgcaa gacgggcctt ttcacgatca tcccagtaac 44040 gcgcccgcgc ttcatcgtta acaaaataat catccttgcg cagattccag atgtcgtctg 44100 ctttcttata cgcagcctct gccttaatca gcatctcctg cgcggtatca ggacgaccaa 44160 tatccagcac cgcatcccac atggatttga atgcccgcgc agtcctgtct gcccaggtct 44220 ccagcgtgcc catgttctct ttcaggcggc gggtctggtc atcaaaccct ttcgttgcgg 44280 cctcgttcgc cgcctgcaat gccccggctt catcgccgga acgctgcaac tgagcaacat 44340 acgcaatctg ctccgccgac acgttatgga actggcgagc catcgccgtc agccccgacg 44400 tcgggtctgt ggtcagcttc ccgaaggctt cagcgacctt tgtccacctc cacgccggat 44460 gcagaggaga aacgcgccac gatcttctag aggcctagaa ggccagagtg gcagagactg 44520 cattcgaaaa cgtttgaatt gataattatt atcatttgcg ggtcaattct tagaaaaact 44580 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt 44640 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggattgcaa 44700 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc 44760 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg 44820 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct 44880 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga 44940 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc 45000 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata 45060 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac 45120 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca 45180 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg 45240 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag 45300 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag 45360 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca 45420 gttttattgt tcatgcgaaa acgtttgaat tgataattat tatcatttgc gggtcctttc 45480 cggcgatccg ccttgttacg gggcggcgac ctcgcgggtt ttcgctattt atgaaaattt 45540 tccggtttaa ggcgtttccg ttcttcttcg tcataactta atgtttttat ttaaaatacc 45600 ctctgaaaag aaaggaaacg acagctgaaa gcgagctttt tggcctctgt cgtttccttt 45660 ctctgttttt gtccgtggaa tgaacaatgg aagtcggcct cgtgatacgc ctatttttat 45720 aggttaatgt catgataata atggtttctt agcgatattt aaattaat 45768

Claims (25)

  1. SFTP(Surfactant Protein) 유전자의 프로모터, 표지 유전자 및 miR 142를 표적하는 염기서열을 포함하는 바이러스 벡터.
  2. 제 1항에 있어서, SFTP 유전자 프로모터는 SFTPA2(Surfactant Protein A2) 유전자 프로모터 또는 SFTPC(Surfactant Protein C) 유전자 프로모터인, 바이러스 벡터.
  3. 제 2항에 있어서, SFTPA2 유전자의 프로모터는 서열번호 5로 표시되는 염기서열을 포함하는, 바이러스 벡터.
  4. 제 2항에 있어서, SFTPAC 유전자의 프로모터는 서열번호 6으로 표시되는 염기서열을 포함하는, 바이러스 벡터.
  5. 제 1항에 있어서, 아데노바이러스 벡터인, 바이러스 벡터.
  6. 제 5항에 있어서, E3 영역 내에 표지 유전자 및 miR 142를 표적하는 염기서열을 포함하는, 바이러스 벡터.
  7. 제 5항에 있어서, E1 영역 내 핵산 서열의 결실을 포함하는, 바이러스 벡터.
  8. 제 5항에 있어서, SFTP 유전자의 프로모터가 아데노바이러스의 내재적 유전자와 작동 가능하게 연결된, 바이러스 벡터.
  9. 제 8항에 있어서, 아데노바이러스의 내재적 유전자는 E1a, E1b, L1, L2, E2, L3, L4, E3 또는 E4를 포함하는, 바이러스 벡터.
  10. 제 1항에 있어서, miR 142를 표적하는 염기서열은 miR-142-3p 표적서열인, 바이러스 벡터.
  11. 제 8항에 있어서, miR-142-3p 표적서열은 서열번호 7로 표시되는 염기서열을 포함하는, 바이러스 벡터.
  12. 제 1항에 있어서, 표지는 발색효소, 방사성 동위원소, 크로모포어(chromopore), 발광물질, 형광물질, 프로브 또는 택(tag)인, 바이러스 벡터.
  13. 제 12항에 있어서, 형광물질은 루시퍼라아제 (luciferase), 증강 녹색 형광 단백질 (enhanced green fluorescent protein, EGFP), 녹색 형광 단백질 (green fluorescent protein, GFP), 노랑 형광 단백질 (Yellow Fluorescent Protein, YFP), 적색 형광 단백질 (Red Fluorescent Protein, RFP) 또는 청색 형광 단백질 (cyan fluorescent protein, CFP)인, 바이러스 벡터.
  14. 제 1항에 있어서, 폐암세포에서 특이적으로 표지 유전자를 발현하는, 바이러스 벡터.
  15. 제 1항에 있어서, 서열번호 8 또는 9로 표시되는 염기서열을 포함하는, 바이러스 벡터.
  16. 제 1항의 바이러스 벡터를 포함하는, 폐암세포 검출용 조성물.
  17. 제 16항에 있어서, 폐암세포는 순환종양세포인, 폐암세포 검출용 조성물.
  18. 제 16항의 폐암세포 검출용 조성물을 포함하는 폐암 진단용 키트.
  19. 제 18항에 있어서, 폐암세포는 순환종양세포인, 폐암 진단용 키트.
  20. 개체로부터 분리된 시료에 제 16항의 폐암세포 검출용 조성물을 처리하는 단계를 포함하는, 폐암세포 검출 방법.
  21. 제 20항에 있어서, 시료는 혈액, 혈청, 혈장, 타액, 복수, 누액, 뇌척수액, 흉수, 활액, 림프, 또는 뇨인, 암세포 검출 방법.혈액, 양수, 혈장, 혈청, 정액, 골수, 뇨, 조직 생검 또는 세포로부터 단백질을 추출한 용액인, 폐암세포 검출 방법.
  22. 제 20항에 있어서, 폐암세포는 순환종양세포인, 폐암세포 검출 방법.
  23. 제 20항에 있어서, 시료 내 표지 유전자 발현 세포가 폐암세포인 것으로 판단하는 단계를 추가로 포함하는, 폐암세포 검출 방법.
  24. 1) 개체로부터 분리된 혈액에 제 16항의 폐암세포 검출용 조성물을 처리하는 단계;
    2) 혈액 내 표지 유전자 발현 세포를 분리하는 단계; 및
    3) 분리한 세포를 배양하는 단계를 포함하는, 순환폐암세포의 분리 및 배양 방법.
  25. 1) 개체로부터 분리된 혈액에 제 16항의 폐암세포 검출용 조성물을 처리하는 단계;
    2) 혈액 내 표지 유전자 발현 세포를 분리하는 단계; 및
    3) 분리한 세포를 배양하는 단계를 포함하는, 폐암의 진단에 필요한 정보를 제공하는 방법.
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KR20140049593A (ko) * 2011-08-23 2014-04-25 도쿠리쯔 교세이 호진 이야쿠 키반 켄큐쇼 제한 증식형 아데노바이러스

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