KR101833984B1 - 밀베마이신을 생산하는 재조합 미생물 및 이를 이용한 밀베마이신 생산 방법 - Google Patents
밀베마이신을 생산하는 재조합 미생물 및 이를 이용한 밀베마이신 생산 방법 Download PDFInfo
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Abstract
밀베마이신을 생산하는 재조합 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis) 균주 및 이를 이용한 밀베마이신 생산 방법이 제공된다.
Description
밀베마이신을 생산하는 재조합 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis) 균주 및 이를 이용한 밀베마이신 생산 방법이 제공된다.
밀베마이신(Milbemycin)은 스트렙토마이세스 하이그로스코피쿠스 아속 아우레오라크리모수스(Streptomyces hygroscopicus subsp . aureolacrimosus; 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus) (NRRL5739 또는 DSM 41911)라고도 칭함), 스트렙토마이세스 시아네오그리세우스 아속 논시아노게누스(Streptomyces cyaneogriseus subsp . noncyanogenus), 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis) 또는 스트렙토마이세스 빙쳉젠시스(Streptomyces bingchenggensis)등으로부터 생산되는 폴리케타이드 계열의 16-원환 매크로라이드 화합물로서 다양한 형태의 생물학적, 화학적 유도체로서 농업 및 동물의약품 용도로 상품화되어 있다.
밀베마이신의 대표적인 예인 밀베멕틴(milbemectin)은 아래 그림과 같은 구조로서, 밀베마이신 A3 와 밀베마이신 A4가 약 30:70 비율로 혼합되어 있는 형태이다.
이 외에도, 밀베마이신 계열로서 밀베멕틴의 화학적 유도체인 레피멕틴(Lepimectin)이 살충제로서 이용되고 있으며, 동물의약품으로서 밀베마이신 옥심(Milbemycin Oxime)과 네마덱틴(Nemadectin)의 화학적 유도체인 목시덱틴(Moxidectin)이 상업화되어 있다.
한편, 아베멕틴(Avermectin)은 밀베마이신과 같이 멕틴계 살충제로 분류되며, 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis) 등으로부터 생산되는 폴리케타이드 계열의 16-원환 매크로라이드 화합물이다. 이들 중 아베멕틴 B1a와 아베멕틴 B1b의 혼합물은 아바멕틴(Abamectin)으로서 응애 등에 대한 살충제로, 아베멕틴의 화학적 유도체인 에마멕틴 벤조에이트(Emamectin benzoate)는 나방 등에 대한 살충제 성분으로 사용되고 있으며, 동물의약품으로서 아베멕틴의 화학적, 생물학적 유도체인 이버멕틴(Ivermectin), 도라멕틴(Doramectin), 셀라멕틴(Selamectin), 에프리노멕틴(Eprinomectin) 등이 상업화되어 있다.
스트렙토마이세스 아베르미틸리스의 아베멕틴 생산성을 향상시키기 위하여 무작위적인 돌연변이법, 트랜스포존(transposon) 돌연변이법 등이 적용되었으며, 이를 효율적으로 스크리닝하기 위한 고속스크리닝법이 개발되었다[Ikeda et al. 1993; Weaden and Dyson 1998; Gao1 et al. 2010; Gao2 et al. 2010; Wang et al. 2010]. 또한, 생산균주의 주요 유전자를 제거하거나 과발현하여 생산성을 증가시키는 연구가 성공적으로 진행되었다[Duong et al. 2009; Li et al. 2010; Zhuo et al. 2010; Qiu et al. 2011; He et al. 2014; Liu et al. 2015]. 이케다 등은 스트렙토마이세스 아베르미틸리스가 외래의 물질합성 유전자를 효과적으로 발현하여 물질을 생산할 수 있음을 밝히고 지놈엔지니어링을 통해 보다 효과적인 발현 균주로 개발하고 있다 [Ikeda et al. 2014]. 또한, 아베멕틴의 경우 현재 대규모의 상업적 생산이 이뤄지고 있어 아베멕틴 생산균주인 스트렙토마이세스 아베르미틸리스가 산업균주로서 보다 유리할 것으로 여겨지고 있다.
밀베마이신의 생산은 스트렙토마이세스 속에 속하는 세균을 배양하고, 균체를 배양액으로부터 분리하여 생산된 밀베마이신을 균체로부터 유기 용매로 추출한 후, 얻어진 목적 화합물을 포함하는 물질을 정제하는 등의 방법에 의해 행해지고 있다. 종래 밀베마이신의 생산에 관한 기술들을 살펴보면, 일본의 Sankyo 사에서 1980년대 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus)가 밀베마이신을 생산하는 것을 발견한 후, 무작위적인 돌연변이 개발 기술을 통해 상업적 수준의 밀베마이신의 생산을 시작하였다. 그러나, 이와 같이 개발된 균주를 이용하여 밀베마이신을 대량생산 할 때 발효조 내의 용존 산소량에 의한 생산성 변화가 상당히 크다는 것이 보고되었고, 개발된 균주의 생산성을 유지시키는 것이 상당히 어렵다는 것이 보고된 바 있다[Okada and Iwamatu. 1997; Ide et al.1993]. 또한 상기 균주외 밀베마이신을 생산하는 스트렙토마이세스 빙쳉젠시스에서 조절 유전자인 nsdA의 삭제를 통하여 밀베마이신 A4의 생산성을 향상시켰다는 보고가 있고[Wang et al. 2009], 스트렙토마이세스 빙쳉젠시스에서 milD를 삭제하여 밀베마이신 A3/A4의 생산성을 향상시켰다는 보고가 있다[Zhang et al. 2013].
그러나, 종래 기술은 원 균주인 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus)를 이용하여 상업적 수준으로 생산하는 경우에 있어서도 대량생산의 어려움 때문에 생산성 향상을 이루기 어려우며, 신규의 균주를 이용하는 경우에 있어서도 산업적 수준의 생산성을 얻기 까지 장기간이 걸린다는 문제가 있다.
Ikeda et al., J. Bacteriol., 175(7): 2077-2078 (1993년 4월)
Weaden and Dyson, Microbiology, 144:1963-1970 (1998년 7월)
Gao et al., Bioresource Technology, 100:4012-4016 (2009년 4월)
Gao1 et al., Appl. Environ. Microbiol., 76(13):4583-4586 (2010년 7월)
Gao2 et al., Appl. Microbiol. Biotechnol., 85(4):1219-1225 (2010년 1월)
Wang et al., J. Appl. Microbiol., 108(3):851-858 (2010년 3월)
Duong et al., J. Microbiol. Biotechnol., 19(2): 136-139 (2009년 2월)
Li et al., Bioresour. Technol., 101(23): 9228-9235 (2010년 12월)
Zhou et al., Proc. Natl. Acad. Sci. (107(25):11250-11254 (2010년 6월)
Qiu et al., Appl. Microbiol. Biotechnol., 92(2):337-345 (2011년 10월)
He et al., Appl. Microbiol. Biotechnol., 98(1):399-409 (2014년 1월)
Liu et al., Appl. Environ. Microbiol., 81(15):5157-5173 (2015년 8월)
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본 발명은 밀베마이신의 산업용 생산 균주로 사용할 정도의 높은 생산성을 가지는 재조합 미생물 및 이를 이용한 밀베마이신 생산 방법과 관련 있다.
일 예는 아베멕틴의 고생산성 균주에서 아베멕틴 생산 유전자의 일부를 밀베마이신 생산 유전자로 치환함으로써 밀베마이신을 고생산하는 재조합 미생물을 제공한다.
다른 예는 밀베마이신 생산 유전자를 포함하는 밀베마이신을 고생산하는 재조합 미생물 제조에 사용 가능한 재조합 벡터를 제공한다.
다른 예는 상기 재조합 미생물을 이용한 밀베마이신 생산 방법을 제공한다.
다른 예는 아베멕틴의 고생산성 균주에서 아베멕틴 생산 유전자의 일부를 밀베마이신 생산 유전자로 치환함으로써 아베멕틴 유도체를 생산하는 재조합 스트렙토마이세스 아베르미틸리스 균주를 제공한다.
다른 예는 신규한 아베멕틴 유도체 또는 이의 염을 제공한다.
다른 예는 상기 신규한 아베멕틴 유도체 또는 이의 염을 포함하는 작물 보호제 및/또는 동물의약용 조성물을 제공한다.
상기 문제를 해결하기 위한 하나의 방안으로서, 본 명세서에서는, 아베멕틴(avermectin)과 밀베마이신(milbemycin)의 분자구조가 매우 유사하고 이들을 생산하는 폴리케타이드 합성효소의 모듈(module) 및 도메인(domain)의 구성이 유사함에 근거하여 아베멕틴 고생산성 균주의 아바멕틴 합성 폴리케타이드 합성효소를 밀베마이신 합성 폴리케타이드 합성효소로 전환함으로서 고생산성의 밀베마이신 생산균주를 제작하는 기술이 제공된다 (도 18a-18d 참조).
하나의 양태로서, 본 발명은 아베멕틴의 고생산성 균주에서 아베멕틴 생산 유전자의 일부를 밀베마이신 생산 유전자로 치환하여 얻어진 재조합 미생물에 관한 것이다. 상기 재조합 미생물은 밀베마이신을 고효율로 생산하는 특징으로 갖는 것일 수 있다.
도 18a-18c는 아베멕틴과 밀베마이신의 분자구조 차이 및 이를 유발하는 유전자의 구성을 보여주는 그림이고, 도 18d는 밀베마이신 폴리케타이드 합성효소 유전자군 (gene cluster)과 아베멕틴 폴리케타이드 합성효소 유전자군을 비교하여 보여주는 그림이다. 도 18a-18d에서, 각 원은 각각 도메인을 나타내며, 각 도메인으로부터 코딩되는 단백질은 각각 다음의 활성을 갖는다 (AT: 아실트랜스퍼라제 (Acyltransferase), KS: 케토-아실기 합성효소 (ketosynthase), KR: 케토-아실기 환원효소 (ketoreductase), DH: 탈수효소 (dehydratase), ER: 엔올 환원효소 (enoyl reductase), ACP: 아실기 운반단백질 (Acyl Carrier Protein), TE: 티오에스터라제 (Thioesterase)). 각각의 합성효소는 모듈로 구성되고 각 모듈은 도메인들로 구성되며, 합성효소 유전자군은 모듈 코딩 유전자들로 구성되며, 각 모듈 코딩 유전자는 각 도메인 코딩 유전자들로 구성된다.
각 모듈 내의 각 도메인은 KS 도메인부터 시작하여 ACP 도메인으로 끝나는 순서로 배열될 수 있다 [예컨대, N-말단에서 C-말단 방향으로, (KS)-(AT)-(DH 및/또는 KR; in any order)-(ACP) 순서로 배열, milA3의 모듈 7의 경우 (KS)-(AT)-(DH)-(ER)-(KR)-(ACP) 순서로 배열]. 밀베마이신 및 아베멕틴 폴리케타이드 합성효소의 모듈 10의 KR 도메인은 폴리케타이드 합성에 관여하지 않으며, 밀베마이신 폴리케타이드 합성효소 모듈 11의 KR 도메인과 아베멕틴 폴리케타이드 합성효소 모듈 7의 DH 도메인은 활성이 없다.
도 18a-18d에서 알 수 있듯이, 아베멕틴과 밀베마이신의 분자 구조가 매우 유사하고, 아베멕틴 폴리케타이드 합성효소 유전자군과 베마이신 폴리케타이드 합성효소 유전자군은 그 구성이 매우 유사하다.
두 화합물(아베멕틴과 밀베마이신)의 합성에 있어 차이는 다음에 기인한다:
1) 폴리케타이드 합성효소의 loading 모듈의 AT의 기질특이성 차이로 밀베마이신 합성효소의 loading 모듈의 경우 acetyl-CoA와 propionyl-CoA를 주요 기질로 사용하고, 아베멕틴 합성효소의 loading 모듈의 경우 isobutyryl-CoA와 2-methylbutanoyl-CoA를 주요기질로 사용하고 있어서, 두 화합물의 각 탄소위치 25번에 연결되어 있는 탄소 사슬의 길이가 달라진다.
2) 두 화합물의 탄소위치 22-23의 탄소결합은 각 합성효소의 모듈 2의 도메인 구성에 의해 결정되어, DH-KR 도메인을 갖는 아베멕틴 폴리케타이드 합성효소 모듈 2의 경우 이중 결합 또는 -OH 결합을 생성하는 반면, DH-ER-KR 도메인을 갖는 밀베마이신 폴리케타이드 합성효소 모듈 2는 단일결합을 생성한다.
3) 두 화합물의 탄소위치 13의 -OH기의 생성은 각 합성효소의 모듈 7의 도메인 구성에 의한다. 아베멕틴 폴리케타이드 합성효소의 경우 KR 도메인만 작용하여 -OH가 생성되지만, 밀베마이신 폴리케타이드 합성효소의 경우 DH-ER-KR 도메인이 있어 탄소 단일결합의 형태가 생성된다.
상기 내용에 기초하여, 아베멕틴 생성 균주를 밀베마이신 생성 균주로 전환시키기 위하여 필수적으로 필요한 최소한의 도메인만을 선택적으로 치환할 수 있다. 본 명세서에서는 도메인 치환시의 활성 유지 정도를 고려하여, 적절한 모듈 또는 도메인 단위를 선택하여 치환을 시도하였다.
보다 구체적으로, aveA3와 milA3의 기능적 차이는 모듈 7의 DH (dehydratase) 와 ER (Enoyl reductase) 활성 도메인의 포함 여부에 의존할 수 있다. aveA3의 모듈은 DH 도메인을 갖고 있지만 활성이 없는 반면, milA3는 모듈 7에 DH와 ER 활성을 갖는 도메인을 포함한다. 이외에, aveA3의 모듈 8과 모듈 9는 milA3의 모듈8과 모듈 9와 동일 내지 유사한 기능을 갖는다. 따라서, 기능적인 측면을 고려하면, 최소한 aveA3의 모듈 7 코딩 유전자 중의 DH 도메인 코딩 유전자를 milA3의 모듈7 코딩 유전자 중의 DH-ER 도메인 코딩 유전자로 치환시킴으로써, 아베멕틴 생성균주에 밀베마이신 생산 활성을 부여할 수 있다. 임의로, 폴리케타이드 합성효소와 같은 거대 효소에 있어 단백질 상호작용에 의해 합성이 조절되는 것이 매우 중요하기 때문에, 단순한 도메인 치환에 의하여 생길 수 있는 활성 감소를 방지하기 위하여, 모듈 단위 (예컨대, 모듈 7)로 치환하거나, 또는 앞서 설명한 도메인 이외의 적절한 도메인 또는 모듈을 추가로 치환할 수 있다.
일 예에서, 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis) 균주에서,
아베멕틴 생합성을 수행하는 폴리케타이드 합성효소 유전자군의 1번 유전자(aveA1; 예컨대, Streptomyces avermitilis MA-4680의 aveA1 유전자 (GeneBank Accession number AB032367.1의 101번째부터 12019번째까지의 폴리뉴클레오타이드 부위(DNA 서열: 서열번호 66; 아미노산 서열 (BAA84474.1): 서열번호 67)) 등)가 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소 유전자군의 1번 유전자(milA1)로 치환되고,
아베멕틴 생합성을 수행하는 폴리케타이드 합성효소 유전자군의 3번 유전자 (aveA3)의 전부 또는 일부가 밀베마이신 생산 균주에서 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소 유전자군의 3번 유전자 (milA3)의 전부 또는 일부로 치환된,
재조합 스트렙토마이세스 아베르미틸리스 균주가 제공된다.
상기 aveA3 일부가 milA3의 일부로 치환된 것은, aveA3 (예컨대, Streptomyces avermitilis MA-4680의 aveA3 유전자의 적어도 모듈 7 코딩 유전자의 전부 또는 일부 (적어도 DH 도메인 코딩 유전자를 포함)가 밀베마이신 생산 균주에서 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소 유전자군의 3번 유전자 (milA3)의 모듈 7 코딩 유전자의 전부 또는 일부 (적어도 DH (dehydratase) 도메인 코딩 유전자 및 ER (enoyl reductase) 도메인 코딩 유전자를 포함)로 치환된 것을 의미할 수 있다.
예컨대, 상기 aveA3는 GeneBank Accession number AB032367.1의 33436번째부터 50034번째까지의 폴리뉴클레오타이드 부위(DNA 서열: 서열번호 71 (polyketide synthase gene cluster (AB032367.1)의 33436번째부터 50034번째까지의 폴리뉴클레오타이드 부위); 아미노산 서열 (BAA84478.1): 서열번호 72 (Module 7: 38번째부터 1841까지 폴리펩티드; Module 8: 1862 번째부터 3641까지 폴리펩티드; Module 9: 3660번째부터5438까지 폴리펩티드; module 7의 DH domain: 976번째부터 1148번째까지 폴리펩티드))일 수 있으며, 상기 aveA3의 일부는 적어도 모듈 7 (BAA84478.1 (서열번호 72)의 35번째부터 1841번째 또는 38번째부터 1841번째까지의 부위)의 코딩 유전자의 전부 또는 일부 (적어도 DH (dehydratase) 도메인 (BAA84478.1 (서열번호 72)의 976번째부터 1148번째까지의 부위)의 코딩 유전자를 포함)을 의미할 수 있다.
aveA3 유전자의 각 모듈, 도메인 및 이의 코딩 유전자는 웹사이트 (http://www.ncbi.nlm.nih.gov/protein/5902891)를 통하여 명확하게 특정할 수 있다.
상기 재조합 스트렙토마이세스 아베르미틸리스 균주는 밀베마이신의 고효율 생산 활성을 갖는 것일 수 있다.
폴리케타이드 합성효소 상에서 도메인, 모듈, 및/또는 단백질의 변이(예컨대, 치환) 시에, 각 영역을 암호화하는 유전자를 연결하는 docking 부분간의 상호작용이 해당 폴리케타이드 합성효소의 정상적 발현에 매우 중요하다. 특히 KS (ketosynthase) 도메인, AT (acyltransferase) 도메인, DH (dehydratase), ER (enoylreductase) 도메인, KR (ketoreductase) 도메인, ACP (acyl carrier protein) 도메인 등을 포함하는 복합효소를 포함하는 폴리케타이드 합성효소 (상기한 도메인들은 N-말단에서 C-말단 방향으로 상기 기재 순서대로 위치하는 것일 수 있다)에서, 각 모듈의 KS와 AT는 upstream의 ACP와 상호작용한다. 이를 고려할 때, 아베멕틴 폴리케타이드 합성효소의 정상적 발현을 위해서, aveA3 유전자의 일부가 milA3 유전자의 일부로 치환 시 치환이 일어나는 염기서열의 위치가 중요하게 고려되어야 한다.
상기 milA3 유전자의 일부로 치환되는 aveA3 유전자의 일부는 적어도 모듈 7의 전부 또는 일부 (적어도 DH 도메인을 포함)의 코딩 유전자를 포함할 수 있고, 이를 치환하는 milA3 유전자의 일부는 적어도 모듈 7의 전부 또는 일부 (적어도 DH 도메인 및 ER 도메인을 포함)의 코딩 유전자를 포함할 수 있다.
aveA3 유전자의 일부를 밀베마이신 생산 균주의 milA3 유전자의 일부로 치환 시, aveA3 유전자의 N-말단 코딩 부위 및 C-말단 코딩 부위의 docking 영역에 더하여, 모듈 7의 KS (KS7), 모듈 7의 AT 도메인 (AT7), 및/또는 모듈 9의 ACP 도메인 (ACP9)의 코딩 유전자 부위들이 아베멕틴 폴리케타이드 합성효소(aveA3 유전자)의 상기 도메인의 코딩 유전자 부위들로 보존되도록 유전자 치환이 일어날 수 있다.
예컨대, aveA3 유전자의 일부 치환은
(a) aveA3 유전자 중 적어도 모듈 7 코딩 유전자 또는 모듈 7의 DH (dehydratase) 도메인 코딩 유전자가 밀베마이신 생산 균주의 milA3 유전자의 모듈 7 코딩 유전자 또는 모듈 7의 DH (dehydratase) 도메인 및 ER (enoyl reductase) 도메인 코딩 유전자로 치환되거나; 또는
(b) 상기 (a) 치환에 더하여, 다음을 추가로 포함하는 치환일 수 있다:
(i) aveA3 유전자의 모듈 7 코딩 유전자 중의 DH 도메인 코딩 유전자를 제외한 나머지 도메인 중 하나 이상의 코딩 유전자 (예컨대, KS 도메인 코딩 유전자, AT 도메인 코딩 유전자, KR 도메인 코딩 유전자 및 ACP 도메인 코딩 유전자로 이루어진 군에서 선택된 하나 이상의 유전자)의 밀베마이신 생산 균주의 milA3 유전자의 모듈 7 코딩 유전자 중의 DH 도메인 및 ER 도메인을 제외한 나머지 도메인 중 하나 이상의 코딩 유전자 (예컨대, KS 도메인 코딩 유전자, AT 도메인 코딩 유전자, KR 도메인 코딩 유전자 및 ACP 도메인 코딩 유전자로 이루어진 군에서 선택된 하나 이상)로의 치환;
(ii) aveA3 유전자의 모듈 8 및 모듈 9의 도메인들 중 하나 이상의 코딩 유전자의 밀베마이신 생산 균주의 milA3 유전자의 모듈 8 및 모듈 9의 도메인들 중 하나 이상의 코딩 유전자로의 치환; 또는
(iii) 치환 (i) 및 (ii)의 조합.
앞서 설명한 aveA1 유전자의 전부가 milA1 유전자의 전부로 치환 및/또는 aveA3 유전자의 전부 또는 일부가 milA3 유전자의 전부 또는 일부로 치환된 균주는, aveA1 및/또는 aveA3 유전자의 유전자 치환 부위의 N-말단 및/또는 C-말단의 docking 영역 코딩 부위가 보존되어 있는 것일 수 있다. 즉, aveA1 유전자의 전부가 milA1 유전자의 전부로 치환 및/또는 aveA3 유전자의 전부 또는 일부가 milA3 유전자의 전부 또는 일부로 치환된 균주는, aveA1 및/또는 aveA3 유전자의 유전자 치환 부위의 upstream 및/또는 downstream 영역과 상동성을 가지는 부위를 상기 치환(도입)된 milA1 유전자 및/또는 milA3 유전자의 전부 또는 일부의 어느 한 말단 또는 양쪽 말단에 연결된 형태로 포함할 수 있다. 이러한 상동성을 가지는 부위의 도입은 상동 재조합시에 유리할 수 있다. 상기 상동성은 원래의 유전자 서열과 90% 이상, 95% 이상, 또는 98% 이상 동일성을 가짐을 의미한다.
상기 사용된 용어 "Docking 영역"은 폴리케타이드 합성효소를 코딩하고 있는 유전자 양 끝 부위에 의하여 코딩되는 부위로, 폴리케타이드 합성시 다음 단계의 합성효소와의 단백질-단백질 상호작용을 원활하게 하는 역할을 하는 부위이다. 통상적인 상동성 염기서열 분석을 범위의 특정이 가능하다.
상기 밀베마이신 생산 균주는 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus) (milA1 유전자: 서열번호 70; milA3 유전자: 서열번호 73; milA3 단백질: 서열번호 74 (BAA84478.1) (Module 7: 34번째부터 2139 번째까지 폴리펩티드; Module 8: 2163번째부터 3927번째까지 폴리펩티드; Module 9: 3951번째부터 5731 번째까지 폴리펩티드: DH-ER domain in module 7: 953번째부터 1775 번째까지 폴리펩티드 (DH domain in module 7: 953번째부터 1129 번째까지 폴리펩티드; ER domain in module 7: 1497 번째부터 1775 번째까지 폴리펩티드))), 스트렙토마이세스 시아네오그리세우스 아속 논시아노게누스(Streptomyces cyaneogriseus subsp . Noncyanogenus; 예컨대, Accession no. CP010849), 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis; 예컨대, Accession no. FJ952082; milA1 유전자 (Meilingmycin이 milbemycin과 구조가 유사하고 기능이 동일하여, 본 명세서에서는 meiA1와 milA1을 동일한 의미로 사용함): 서열번호 69; milA3 유전자(본 명세서에서는 meiA3와 milA3을 동일한 의미로 사용함): 서열번호 75 (GenBank: FJ952082 Sequence 영역 78606번째부터 96074번째까지 폴리뉴클레오티드); milA3 단백질: 서열번호 76 (Module 7: 39번째부터2143 번째까지 폴리펩티드; Module 8: 2166번째부터 3931 번째까지 폴리펩티드; Module 9: 3952번째부터 5734 번째까지 폴리펩티드; Module 7의 DH-ER didomain 957번째부터 2143 번째까지 폴리펩티드 (Module 7의 DH domain: 957번째부터 1133 번째까지 폴리펩티드; 모듈 7의 ER domain: 1501번째부터 1779 번째까지 폴리펩티드))) 또는 스트렙토마이세스 빙첸젠시스(Streptomyces bingchenggensis; 예컨대, Accession no. CP002047.1; milA1 유전자: 서열번호 68; milA3 유전자: 서열번호 77 (Accession no. CP002047.1 중 1063754::1081234 (유전자가 3'->5' 이어서 reverse complement seq를 기재함)); 단백질: 서열번호 78 (Genbank Accession No. ADI03854; Module 7: 35::2150, Module 8: 2173::3938, Module 9: 3990::5738, DH-ER didomain in module 7: 950::1772 (DH domain: 950::1126; ER domain: 1494::1772)))일 수 있다.
다른 구체예로, 상기 재조합 균주는 밀베마이신 A3, 밀베마이신 A4, 밀베마이신 α2, 밀베마이신 α4 및 밀베마이신 G 로 이루어진 군에서 선택되는 1종 이상의 밀베마이신을 생산할 수 있다.
다른 구체예로, 상기 재조합 균주는 스트렙토마이세스 아베르미틸리스 DBM-03-A 균주(수탁번호 KCTC12890BP), 스트렙토마이세스 아베르미틸리스 DBM-03-B 균주(수탁번호 KCTC12891BP), 및 스트렙토마이세스 아베르미틸리스 DBM-03-C 균주(수탁번호 KCTC13083BP)로 이루어진 군에서 선택된 1종 이상일 수 있다.
다른 양태로서, 앞서 설명한 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소 유전자 milA1 및 milA3의 전부 또는 일부 (적어도 모듈 7 코딩 유전자 또는 모듈 7의 DH-ER 도메인 코딩 유전자를 포함)를 포함하는 재조합 벡터를 적절한 숙주세포 (예컨대, 스트렙토마이세스 아베르미틸리스와 같은 아베멕틴 생성 균주)에 도입하는 단계를 포함하는 재조합 미생물 (예컨대, 재조합 스트렙토마이세스 아베르미틸리스 균주)의 제조 방법을 제공한다.
상기와 같이 제조된 재조합 미생물은 상기 재조합 벡터가 도입되지 않은 미생물과 비교하여 밀베마이신을 생산 효율이 현저히 증가한 특징을 갖는다.
상기 도입시에, 숙주세포 (예컨대, 스트렙토마이세스 아베르미틸리스와 같은 아베멕틴 생성 균주)의 폴리케타이드 합성효소 유전자 aveA1 및 aveA3의 전부 또는 일부 (적어도 모듈 7 코딩 유전자 또는 모듈 7의 DH 도메인 코딩 유전자를 포함)가 milA1 및 milA3의 전부 또는 일부 (적어도 모듈 7 코딩 유전자 또는 모듈 7의 DH-ER 도메인 코딩 유전자를 포함)로 치환이 일어날 수 있다. 또한, aveA1 및 aveA3의 활성이 제거된 상태에서 milA1 및 milA3의 전부 또는 일부 (적어도 모듈 7 코딩 유전자 또는 모듈 7의 DH-ER 도메인 코딩 유전자를 포함)를 포함하는 재조합 벡터가 도입되는 경우 보다 상승된 밀베마이신 생산 효과를 얻을 수 있다.
또 하나의 양태로서, 상기 재조합 미생물 (즉, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주) 또는 상기 재조합 벡터를 포함하는 밀베마이신 제조용 조성물이 제공된다.
또 하나의 양태로서, 상기 재조합 미생물을 이용한 밀베마이신 생산 방법이 제공된다.
구체예로, 본 발명은 상기 재조합 스트렙토마이세스 아베르미틸리스 균주를 배양하는 단계, 및 상기 배양된 균주 또는 균주의 배양물로부터 밀베마이신을 수득하는 단계를 포함하는, 밀베마이신을 생산하는 방법에 관한 것이다.
또 하나의 양태로서, 본 발명은 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis) 균주에서 아베멕틴 생합성을 수행하는 폴리케타이드 합성효소의 1번 유전자(aveA1)가 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 1번 유전자(milA1)로 치환되며, 22,23-디하이드로-25-이소프로필-아베멕틴(22,23-dihydro-25-isopropyl-avermectin), 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin), 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin) 및 5-메톡시-22,23-디하이드로-25-이소프로필-아베멕틴(5-methoxy-22,23-dihydro-25-isopropyl-avermectin)으로 이루어진 군에서 선택되는 1종 이상의 아베멕틴 유도체를 생산하는 재조합 스트렙토마이세스 아베르미틸리스 균주에 관한 것이다. 예컨대, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주는 앞서 설명한 milA1을 포함하는 재조합 벡터를 사용하여 제작된 것일 수 있다.
구체 예로, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주는, 스트렙토마이세스 아베르미틸리스 DBM-01 균주(수탁번호 KCTC12889BP)일 수 있다.
또 하나의 양태로서, 본 발명은 하기에서 선택되는, 아베멕틴 유도체 또는 이의 염에 관한 것이다: 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin) 또는 이의 염, 및 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin) 또는 이의 염.
또 하나의 양태로서, 본 발명은 상기 아베멕틴 유도체 또는 이의 염의 작물 보호제 및/또는 동물의약품으로서의 용도에 관한 것이다.
이하, 본 발명을 보다 상세하게 설명한다.
스트렙토마이세스 아베르미틸리스는 아베멕틴을 생산하는 균주로서, 그 예로는, S. avermitilis SA-01 균주, S. avermitilis MA-4680 균주(NCBI 등록번호: NC_003155.4), S. avermitilis 76-02-e 균주(He et al. 2014), S. avermitilis 14-12A 균주(Gao et al. 2009), S. avermitilis 3-115 균주(Gao et al. 2010) 등을 들 수 있으나, 이에 제한되는 것은 아니다.
스트렙토마이세스 아베르미틸리스는 아베멕틴을 생산하기 위한 PKS 클러스터(Polyketide synthase gene cluster; 폴리케타이드 합성효소 유전자 클러스터)라는 거대 유전자군을 포함하고 있다. 상기 아베멕틴 PKS 유전자군은 aveA1, aveA2, aveA3 및 aveA4 유전자를 포함하고 있으며, aveA1 유전자는 로딩 모듈과 모듈 1번과 모듈 2번, aveA2 유전자는 모듈 3번 내지 모듈 6번, aveA3 유전자는 모듈 7 내지 모듈 9, aveA4 유전자는 모듈 10번 내지 모듈 12번을 포함하고 있고, 각 모듈은 하위 도메인들로 구성되어 있다.
밀베마이신 생산 균주는 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus), 스트렙토마이세스 시아네오그리세우스 아속 논시아노게누스(Streptomyces cyaneogriseus sp . Noncyanogenus), 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis) 또는 스트렙토마이세스 빙첸젠시스(Streptomyces bingchenggensis)를 포함하나, 이에 제한되는 것은 아니다. 스트렙토마이세스 하이그로스코피쿠스 아속 아우레오라크리모수스로는, 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus) NRRL 5739 균주를 사용할 수 있으나, 이에 제한되는 것은 아니다.
밀베마이신 생산 균주는 밀베마이신을 생산하기 위한 PKS 클러스터 유전자군을 포함하고 있다. 상기 밀베마이신 PKS 유전자군의 구성은 아베멕틴 PKS 유전자군의 구성과 유사하며, milA1, milA2, milA3 및 milA4 유전자를 포함하고 있으며, 각 유전자는 모듈 및 하위 도메인들로 구성되어 있다.
본 발명은 아베멕틴 생산 균주인 스트렙토마이세스 아베르미틸리스의 aveA1 유전자, 이에 상응하는 밀베마이신 생산 균주의 milA1 유전자로 치환시킴으로써 스트렙토마이세스 아베르미틸리스 내에 하이브리드 PKS 유전자가 포함되도록 하였고, 상기 하이브리드 PKS 를 포함하는 재조합 스트렙토마이세스 아베르미틸리스 균주가 다양한 아베멕틴 유도체들을 생산할 수 있음을 확인하였다.
상기 재조합 스트렙토마이세스 아베르미틸리스 균주가 생산하는 아베멕틴 유도체는, 22,23-디하이드로-25-이소프로필-아베멕틴(22,23-dihydro-25-isopropyl-avermectin), 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin), 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin) 및 5-메톡시-22,23-디하이드로-25-이소프로필-아베멕틴(5-methoxy-22,23-dihydro-25-isopropyl-avermectin)으로 이루어진 군에서 선택되는 1종 이상을 포함한다.
또한, 이 중에서 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin), 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin)은 종래 구축된 화합물 데이터베이스에서 검색되지 않는 신규한 화합물임이 확인되었다. 이들 신규 아베멕틴 유도체들은 작물 보호제 및/또는 동물의약품 용도로 사용될 수 있으며, 종래 개발된 아베멕틴 유도체들을 대체할 수 있다.
다른 예로, 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis)의 aveA1 유전자가 밀베마이신 생산 균주의 milA1 유전자의 전부 또는 일부로 치환되며, 아베멕틴 유도체들을 생산하는 재조합 스트렙토마이세스 아베르미틸리스 균주가 제공된다.
다른 예로, 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin) 또는 이의 염, 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin) 또는 이의 염, 및 이들 화합물들의 작물 보호제 및/또는 동물의약품으로서의 용도가 제공된다.
상기 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴은 다음의 화학식으로 나타낼 수 있다:
또한, 상기 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴은 다음의 화학식으로 나타낼 수 있다:
또한, 앞서 설명한 바와 같이, 아베멕틴 생산 균주인 스트렙토마이세스 아베르미틸리스의 aveA1 유전자 및 aveA3 유전자의 전체 또는 일부 (적어도 aveA3의 모듈 7의 DH 도메인 코딩 유전자를 포함)를, 이에 상응하는 밀베마이신 생산 균주의 milA1 및 milA3 유전자의 전체 또는 일부 (적어도 milA3의 모듈 7의 DH-ER 도메인 코딩 유전자를 포함)로 치환시킴으로써 스트렙토마이세스 아베르미틸리스 내에 하이브리드 PKS 유전자가 포함되도록 하였고, 상기 하이브리드 PKS 유전자를 포함하는 재조합 균주가 밀베마이신을 생산할 수 있음이 확인되었다.
따라서, 다른 예로, 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis)의 aveA1 유전자의 전부 또는 일부가 밀베마이신 생산 균주의 milA1 유전자의 전부 또는 일부로 치환되고, aveA3 유전자의 전부 또는 일부(적어도 aveA3의 모듈 7의 DH 도메인 코딩 유전자를 포함)가 밀베마이신 생산 균주의 milA3 유전자의 전부 또는 일부(적어도 milA3의 모듈 7의 DH-ER 도메인 코딩 유전자를 포함)로 치환된, 밀베마이신을 생산하는 재조합 스트렙토마이세스 아베르미틸리스 균주가 제공된다.
상기 aveA3 유전자는 전체가 milA3 유전자로 치환될 수도 있으나, milA3 유전자 중에서 숙주 균주에 밀베마이신을 생산능을 부여할 수 있는 최소한의 부위만 aveA3 유전자의 상응하는 부위에 치환(삽입)되는 것이 가능하다. 예를 들어, 상기 milA3 유전자 중에서 숙주 균주에 밀베마이신을 생산능을 부여할 수 있는 최소한의 부위는 milA3 유전자 중 모듈 7 전부 또는 모듈 7의 DH-ER 도메인의 코딩 유전자를 포함할 수 있으며, aveA3 유전자 중 모듈 7 전부 또는 적어도 모듈 7의 DH 도메인을 포함하는 모듈 7의 일부의 코딩 유전자가, 밀베마이신 생산 균주의 milA3 유전자 중 모듈 7 전부 또는 모듈 7의 DH-ER 도메인의 코딩 유전자로 치환될 수 있다.
일 구현예로, 스트렙토마이세스 아베르미틸리스 균주에서 aveA1 유전자가 밀베마이신 생산 균주의 milA1 유전자로 치환된 균주는, 스트렙토마이세스 아베르미틸리스 균주는, 스트렙토마이세스 아베르미틸리스 DBM-01 균주(수탁번호 KCTC12889BP)일 수 있다.
다른 예로, 스트렙토마이세스 아베르미틸리스 균주에서 aveA1 유전자가 밀베마이신 생산 균주의 milA1 유전자로 치환되고, aveA3 유전자의 모듈 7의 코딩 유전자 부위가 밀베마이신 생산 균주의 milA3 유전자의 모듈 7의 코딩 유전자 부위로 치환된 균주는, 스트렙토마이세스 아베르미틸리스 DBM-03-A 균주(수탁번호 KCTC12890BP)일 수 있다.
다른 예로, 스트렙토마이세스 아베르미틸리스 균주에서 aveA1 유전자가 밀베마이신 생산 균주의 milA1 유전자로 치환되고, aveA3 유전자의 모듈 7 중 KS 도메인을 제외한 모듈 7의 도메인들의 코딩 유전자 부위가 밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중 KS 도메인을 제외한 모듈 7의 도메인들의 코딩 유전자 부위로 치환된 균주는, 스트렙토마이세스 아베르미틸리스 DBM-03-B 균주(수탁번호 KCTC12891BP)일 수 있다.
다른 예로, 스트렙토마이세스 아베르미틸리스 균주에서 aveA1 유전자가 밀베마이신 생산 균주의 milA1 유전자로 치환되고, aveA3 유전자의 모듈 7 중 KS 도메인 및 AT를 제외한 모듈 7의 도메인들의 코딩 유전자 부위가 밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중 KS 도메인 및 AT 도메인을 제외한 모듈 7의 도메인들의 코딩 유전자 부위로 치환되고, aveA3 유전자의 모듈 9 중 ACP 도메인을 제외한 모듈 9의 도메인들의 코딩 유전자 부위가 밀베마이신 생산 균주의 milA3 유전자의 모듈 9 중 ACP 도메인을 제외한 모듈 9의 도메인들의 코딩 유전자 부위로 치환된 균주가 제공된다. 상기 균주는 aveA3의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인까지의 영역 (aveA3의 모듈 7의 DH 도메인, KR 도메인, 및 ACP 도메인, 모듈 8, 모듈 9의 KS 도메인, AT 도메인, DH 도메인, 및 KR 도메인을 N-말단에서 C-말단 방향으로 포함하는 영역)의 코딩 유전자 부위가 밀베마이신 생산 균주의 milA3의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인까지의 영역(milA3의 모듈 7의 DH 도메인, ER 도메인, KR 도메인, 및 ACP 도메인, 모듈 8, 모듈 9의 KS 도메인, AT 도메인, DH 도메인, ER 도메인, 및 KR 도메인을 N-말단에서 C-말단 방향으로 포함하는 영역)의 코딩 유전자 부위로 치환된 것일 수 있으며, 예컨대, 스트렙토마이세스 아베르미틸리스 DBM-03-C 균주(수탁번호 KCTC13083BP)일 수 있다.
그러나, 상기 기탁 균주들은 본 발명의 대표적인 구현예에 불과하며, 본 발명의 범위가 이에 제한되는 것은 아니다.
스트렙토마이세스 아베르미틸리스의 aveA1 및/또는 aveA3 유전자의 치환은, 당업계에 알려진 공지의 기술들, 예를 들어 상동 재조합((homologous recombination)에 의하여 수행될 수 있다.
일 구현예로, 밀베마이신 생산 균주에서 분리한 milA1 및/또는 milA3 유전자의 전체 또는 일부가 상동 재조합을 통하여 숙주 균주 게놈에 통합될 수 있도록 하기 위한 유전자 치환용 벡터를 제조할 수 있다. 상기 벡터는 숙주 게놈의 특정 유전자 위치로 목적하는 유전자를 제거 또는 삽입할 수 있는 벡터로, 상동 재조합이 일어나도록 타겟팅하고자 하는 특정 유전자 부위에 상동인 염기 서열을 포함할 수 있다.
본 발명의 일 양태로서, 상기 스트렙토마이세스 아베르미틸리스 재조합 균주 생산에 사용되는 재조합 벡터가 제공된다.
상기 재조합 벡터는
(1) 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 1번 유전자(milA1); 및
(2) 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 3번 유전자(milA3)의 전부 또는 일부 (적어도 milA3 유전자 중 모듈 7 코딩 유전자 또는 모듈 7의 DH 도메인 코딩 유전자 및 ER 도메인 코딩 유전자를 포함)
을 포함하는 것일 수 있다. 이 경우 milA1 유전자와 milA3의 전부 또는 일부는 하나의 벡터에 함께 포함되거나 각각 별개의 벡터에 포함될 수 있다.
일 구체예에서, 상기 재조합 벡터는 밀베마이신 생산 균주의 milA1 유전자; 및 밀베마이신 생산 균주의 milA3 유전자 또는 밀베마이신 생산 균주의 milA3의 모듈 7의 전부 또는 일부의 코딩 유전자에 더하여, 다음으로 이루어진 군에서 선택된 하나 이상을 추가로 포함할 수 있다:
밀베마이신 생산 균주의 milA3의 모듈 7의 DH-ER 도메인을 제외한 도메인들 (예컨대, KS, AT, KR, 및 ACP 도메인) 중에서 선택된 하나 이상의 코딩 유전자;
밀베마이신 생산 균주의 milA3의 모듈 8의 코딩 유전자 또는 모듈 8의 도메인들 중 하나 이상의 코딩 유전자; 및
밀베마이신 생산 균주의 milA3의 모듈 9의 코딩 유전자 또는 모듈 9의 도메인들 중 하나 이상의 코딩 유전자.
일 구체예에서, 상기 재조합 벡터는
밀베마이신 생산 균주의 milA1 유전자; 및 다음에서 선택된 milA3 유전자 일부 또는 이의 변이체를 포함할 수 있다:
밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중의KS 도메인의 코딩 유전자 부위가 결실된, 예컨대, milA3 유전자의 모듈 7의AT 도메인, DH 도메인, ER 도메인, KR 도메인 및 ACP 도메인의 코딩 유전자를 포함하거나 이들로 구성된 milA3 유전자의 일부;
밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중의 KS 도메인의 코딩 유전자가 aveA3 유전자의 모듈 7의 KS 도메인의 코딩 유전자로 치환된 milA3의 모듈 7코딩 유전자 변이체;
밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중의 AT 도메인의 코딩 유전자 부위가 결실된, 예컨대, milA3 유전자의 모듈 7의KS 도메인, DH 도메인, ER 도메인, KR 도메인 및 ACP 도메인의 코딩 유전자를 포함하는 milA3의 모듈 7코딩 유전자 변이체;
밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중의 AT 도메인의 코딩 유전자가 aveA3 유전자의 모듈 7의 AT 도메인의 코딩 유전자로 치환된 milA3의 모듈 7의 코딩 유전자 변이체;
밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중의 KS 도메인 및 AT 도메인의 코딩 유전자가 결실된, 예컨대, milA3 유전자의 모듈 7의 DH 도메인, ER 도메인, KR 도메인 및 ACP 도메인의 코딩 유전자를 포함하는 milA3의 모듈 7의 코딩 유전자 변이체; 및
밀베마이신 생산 균주의 milA3 유전자의 모듈 7 중의 KS 도메인 및 AT 도메인의 코딩 유전자가 각각 aveA3 유전자의 모듈 7의 KS 도메인 및 AT 도메인의 코딩 유전자로 치환된 milA3의 모듈 7의 코딩 유전자 부위 변이체.
상기 재조합 벡터는 다음으로 이루어진 군에서 선택된 하나 이상을 추가로 포함할 수 있다:
밀베마이신 생산 균주의 milA3 유전자의 모듈 9 중의 ACP 도메인의 코딩 유전자가 결실된, 예컨대, milA3 유전자의 모듈 9의 KS 도메인, AT 도메인, DH 도메인, 및 KR 도메인의 코딩 유전자를 포함하는 milA3의 모듈 9의 코딩 유전자 변이체; 및
밀베마이신 생산 균주의 milA3 유전자의 모듈 9 중의 ACP 도메인의 코딩 유전자가 aveA3 유전자의 모듈 9의 ACP 도메인의 코딩 유전자로 치환된 milA3의 모듈 9의 코딩 유전자 변이체.
일 예에서, 상기 재조합 벡터에 포함되는 milA3 유전자 변이체는
밀베마이신 생산 균주의 milA3 유전자 중의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인까지의 영역(예컨대, milA3의 모듈 7의 DH 도메인, ER 도메인, KR 도메인, 및 ACP 도메인, 모듈 8, 모듈 9의 KS 도메인, AT 도메인, DH 도메인, ER 도메인, 및 KR 도메인을 N-말단에서 C-말단 방향으로 포함하는 영역)의 코딩 유전자를 포함하는 milA3 유전자 변이체 (일부)일 수 있으나 이에 제한되는 것은 아니다.
본원의 일 실시예에서, milA1을 치환하기 위한 재조합 벡터는 milA1 유전자 전체를 포함하고, 상동 재조합을 위하여 aveA1 유전자의 upstream 및 downstream 지역과 상동성을 가지는 부위를 milA1의 어느 한쪽 말단 또는 양쪽 말단에 연결된 형태로 구성될 수 있다. 또한, 다른 실시예에서는, milA3을 치환하기 위한 재조합 벡터는, milA3 유전자의 7번 모듈의 코딩 유전자의 전체 또는 일부를 포함하고, 상동 재조합을 위하여 aveA3 및/또는 aveA4 유전자 지역과 상동성을 가지는 부위를 milA3 유전자의 7번 모듈의 코딩 유전자의 전체 또는 일부의 어느 한쪽 말단 또는 양쪽 말단에 연결된 형태로 구성될 수 있다. 상기 벡터는 본원의 대표적인 실시예일뿐이며, 본원의 범위가 이에 제한되는 것은 아니다.
상기 벡터는, 상동 재조합을 위하여 숙주 유전자 부위와 상동성을 가지는 부위를 포함하는데, 여기에서 상동이란 숙주 유전자 부위의 염기 서열과의 동일성 정도를 나타내는 것으로, 예를 들어, 숙주 유전자의 염기서열과 90% 이상, 95% 이상 또는 98% 이상 동일한 것일 수 있다.
스트렙토마이세스에서 사용하기 위한 다양한 여러 벡터들, 예를 들어 파지, 고 카피수(high copy number) 플라스미드, 저 카피수 플라스미드, 및 대장균(E. Coli)-스트렙토마이세스 셔틀 벡터들이 개발되어 왔으며, 이들 벡터들을 본 발명을 구현하기 위하여 사용할 수 있다. 예를 들어, pCR-Blunt, pCR2.1(Invitrogen), pGEM3Zf(Promega) 및 셔틀 벡터 pWHM3, pKC1139 등을 예시할 수 있으나, 이에 제한되는 것은 아니다.
상기 벡터는, 형질전환된 세포를 선별하기 위한 목적으로 선별마커(selection marker)를 더욱 포함할 수 있으며, 예를 들어, 약물 내성, 영양 요구성, 세포 독성제에 대한 내성 또는 표면 단백질의 발현과 같은 선택가능 표현형을 부여하는 마커들이 사용될 수 있고, 포지티브 선별마커와 네가티브 선별마커를 예시할 수 있다. 포지티브 선별마커란 선택제(selective agent)가 처리된 환경에서 선택 마커를 발현하는 세포만 생존하도록 하여 포지티브 선택을 가능하게 하는 마커로, 아프라마이신, 네오마이신, 하이그로마이신, 히스티디놀 디하이드로게나제(histidinol dehydrogenase gene: hisD) 또는 구아닌 포스포리보실트랜스퍼라제(guanine phosphosribosyltransferase: Gpt) 등이 있으나, 이에 제한되지 않는다. 네가티브 선별마커란 무작위적 삽입(random insertion)이 일어난 세포를 선별하여 제거하는 네가티브 선택을 가능하게 하는 마커로, 허피스 심플렉스 바이러스-싸이미딘 키나제(Herpes simplex virus-thymidine kinase: HSV-tk), 하이포잔틴 포스포리보실 트랜스퍼자제(hypoxanthine phosphoribosyl transferase: Hprt), 싸이토신 디아미네즈(cytosine deaminase), 디프테리아 톡신(Diphtheria toxin) 등이 있으나, 이에 제한되지 않는다.
상기 벡터 제작은 당해 기술분야에서 잘 알려진 유전자 재조합 기술을 이용하여 제조할 수 있으며, 부위-특이적 DNA 절단 및 연결은 당해 기술 분야에서 일반적으로 알려진 제한효소 등을 사용할 수 있다.
본 발명의 벡터는 스트렙토마이세스 세포에서 작용하지만, 예를 들어 클로닝 또는 발현 목적을 위해 다른 박테리아 또는 진핵 세포로 또한 형질전환될 수 있다. 예를 들어, 아메리칸 타입 컬쳐 컬렉션(American Type Culture Collection; ATCC)로부터 구입할 수 있거나, 시판 중인 DH5α 균주와 같은 에스케리키아 콜라이 균주가 전형적으로 사용될 수 있다. 바람직한 진핵 숙주 세포로는 포유동물 세포, 곤충 세포 또는 효모 세포가 또한 효과적으로 사용될 수 있다.
숙주 균주 내로 본 발명의 벡터를 도입 방법은 핵산을 세포 내로 도입하는 어떠한 방법도 포함되며, 당 분야에서 공지된 바와 같이 적합한 표준 기술을 선택하여 수행할 수 있다. 예들 들어, 원형질체(protoplast) 형질전환, 전기천공법(electroporation), 전기주입법(electroinjection), 미세주입법 (microinjection), 인산칼슘공동-침전법(calcium phosphate co-precipitation), 염화캄슘/염화루비듐법, 레트로바이러스 감염(retroviral infection), DEAE-덱스트란(DEAE-dextran), 양이온 리포좀(cationic liposome)법, 폴리에틸렌글리콜 침전법(polyethylene glycol-mediated uptake), 유전자총(gene gun) 등을 이용할 수 있으나, 이에 제한되는 것은 아니다. 이 때 원형의 벡터를 적절한 제한효소로 절단하여 선형의 벡터 형태 또는 플라스미드를 제거한 선형의 벡터 형태로 도입할 수 있다. 형질전환체는 선별마커, 예를 들어 전술한 바와 같이 재조합 벡터와 연관된 항생물질 저항성을 발현시키는 세포를 선택하는 방법과 같은 표준 절차에 따라 선택할 수 있다.
이와 같이 제조된 재조합 균주를 배양하여 밀베마이신을 생산할 수 있으며, 예를 들어, 밀베마이신 A3, 밀베마이신 A4, 밀베마이신 α2, 밀베마이신 α4 및 밀베마이신 G 로 이루어진 군에서 선택되는 1종 이상의 밀베마이신을 생산할 수 있다.
재조합 균주의 배양은 균주의 생육과 밀베마이신의 대량 생산에 적합하도록 온도, 배지의 pH 및 배양시간 등의 조건들을 적절하게 조절할 수 있다. 상기 배양 방법의 예에는, 회분식, 연속식 및 유가식 배양이 포함되나, 이에 제한되는 것은 아니다.
배양에 사용되는 배지는 특정한 균주의 요구조건을 적절하게 만족시켜야 한다. 상기 배지는 다양한 탄소원, 질소원, 인원 및 미량원소 성분을 포함할 수 있다. 발현 벡터가 유도성 촉진자를 포함하는 경우에는, 온도 변화, 영양원의 고갈, 무상 유도 물질(예를 들어, 이소프로필-β-D-티오갈락토피라노시드(IPTG)와 같은 탄수화물의 유사 물질)의 첨가, 과잉 대사 부산물의 축적 등과 같은 적절한 유도 조건은 발현을 유도하는데 필요에 따라 적용될 수 있다.
배지 내 탄소원으로는 글루코즈, 사카로즈, 락토즈, 프락토즈, 말토즈, 전분, 셀룰로즈와 같은 당 및 탄수화물, 대두유, 해바라기유, 피마자유, 코코넛유 등과 같은 오일 및 지방, 팔미트산, 스테아린산, 리놀레산과 같은 지방산, 글리세롤, 에탄올과 같은 알코올, 아세트산과 같은 유기산을 예시할 수 있으나, 이에 제한되는 것은 아니다. 이들 물질은 개별적으로 또는 혼합물로서 사용될 수 있다. 배지 내 질소원으로는 펩톤, 효모 추출물, 육즙, 맥아 추출물, 옥수수 침지액, 대두밀 및 요소 또는 무기 화합물, 예를 들면 황산암모늄, 염화암모늄, 인산암모늄, 탄산암모늄 및 질산암모늄을 예시할 수 있으나, 이에 제한되는 것은 아니다. 질소원 또한 개별적으로 또는 혼합물로서 사용할 수 있다. 배지 내 인원으로는 인산이수소칼륨 또는 인산수소이칼륨 또는 상응하는 나트륨-함유 염을 예시할 수 있으나, 이에 제한되는 것은 아니다. 또한, 배양 배지는 성장에 필요한 황산마그네슘 또는 황산철과 같은 금속염을 포함하거나, 아미노산 및 비타민과 같은 필수 성장 물질을 포함할 수 있으나, 이에 제한되는 것은 아니다. 상기된 원료들은 배양 과정에서 배양물에 적절한 방식에 의해 회분식으로 또는 연속식으로 첨가될 수 있다.
또한, 필요에 따라, 수산화나트륨, 수산화칼륨, 암모니아와 같은 기초 화합물 또는 인산 또는 황산과 같은 산 화합물을 적절한 방식으로 사용하여 배양물의 pH를 조절할 수 있다. 또한, 지방산 폴리글리콜 에스테르와 같은 소포제를 사용하여 기포 생성을 억제할 수 있다. 호기 상태를 유지하기 위해 배양물 내로 산소 또는 산소-함유 기체(예, 공기)를 주입할 수 있으며, 배양물의 온도는 보통 20 내지 45, 바람직하게는 25 내지 40 일 수 있다. 배양은 원하는 밀베마이신의 생산량이 최대로 얻어질 때까지 계속될 수 있다.
재조합 미생물로부터 생산된 밀베마이신은, 당업계에 널리 알려져 있는 방법으로 세포, 세포 용해물 또는 배양 배지로부터 단리되거나 실질적으로 정제될 수 있다. 밀베마이신의 회수 방법의 예로서, 유기용매 추출법, 원심분리, 초음파파쇄, 여과, 결정법(crystallization), 이온교환 크로마토그래피, 친화성 크로마토그래피, 고성능 액체 크로마토그래피(high performance liquid chromatography: HPLC), 가스 크로마토그래피(gas chromatography: GC) 등의 방법이 있으나, 이들 예에 한정되는 것은 아니다. 구체예로, 균주 배양물로부터 유기용매를 이용하여 생성 물질들을 추출한 후, 실리카겔, 알루미나, 덱스트란겔, 이온교환 수지, 합성흡착제, 분자체, C8H17, C18H37, C6H5 등의 화학 결합형 실리카겔 등의 담체를 사용한 크로마토그래피에 부여하여, 얻어진 목적 화합물을 함유하는 분획을 농축 건조시키는 방법으로 회수할 수 있다.
본 발명의 다른 예는 앞서 설명한 재조합 스트렙토마이세스 아베르미틸리스 균주 또는 상기 균주의 배양물; 재조합 벡터; 및 아베멕틴 유도체 또는 이의 염으로 이루어진 군에서 선택된 1종 이상을 유효성분으로 포함하는 해충 방제용 조성물을 제공한다.
다른 예는 앞서 설명한 재조합 스트렙토마이세스 아베르미틸리스 균주 또는 상기 균주의 배양물; 재조합 벡터; 및 아베멕틴 유도체 또는 이의 염으로 이루어진 군에서 선택된 1종 이상을 작물에 적용하는 단계를 포함하는 해충 방제 방법을 제공한다.
상기 해충 방제는 해충에 대한 살충, 기피 등의 저해 효과를 포괄적으로 의미한다.
상기 해충은 동물 및/또는 식물에 위해를 가하는 모든 절지 동물의 성충, 유충 및 알을 총칭하는 것으로, 밀베마이신 또는 이의 유도체, 및/또는 아베멕틴 또는 이의 유도체가 방제 활성을 갖는 모든 해충의 성충 및/또는 유충일 수 있다. 예컨대, 상기 해충은 다음으로 이루어진 군에서 선택된 1종 이상일 수 있다:
- 반시류 (Hemiptera) (예컨대, 다음을 포함함:
멸구류 (Delphacidae), 예컨대, 애멸구 (Laodelphax striatellus), 벼멸구 (Nilaparvata lugens), 흰등멸구 (Sogatella furcifera) 등; 매미충류 (Deltocephalidae), 예컨대, 끝동매미충 (Nephotettix cincticeps), 두점끝 동매미충 (Nephotettix virescens) 등; 진딧물류 (Aphididae), 예컨대, 목화진딧물 (Aphis gossypii), 복숭아혹진 딧물 (Myzus persicae), 양배추가루진딧물 (Brevicoryne brassicae), 감자수염진딧물 (Macrosiphum euphorbiae), 싸리수염진딧물 (Aulacorthum solani), 기장테두리진딧물 (Rhopalosiphum padi), 귤소리진딧물 (Toxoptera citricidus) 등; 노린재류 (Pentatomidae), 예컨대, 풀색노린재 (Nezara antennata), 톱다리개미허리노 린재 (Riptortus clavetus), 호리허리노린재 (Leptocorisa chinensis), 가시점둥글노린재 (Eysarcoris parvus), 썩덩나무노린재 (Halyomorpha mista), 장님노린재 (Lyus lineolaris) 등; 가루이류 (Aleyrodidae), 예 컨대 온실가루이 (Trialeurodes vaporariorum), 담배가루이 (Bemisia tabaci), 은빛잎가루이 (Bemisia argentifolii) 등; 깍지벌레류 (Coccidae), 예컨대, 캘리포니아붉은깍지벌레 (Aonidiella aurantii), 샌호제깍지벌 레 (Comstockaspis perniciosa), 화살깍지벌레 (Unaspis citri), 루비깍지벌레 (Ceroplastes rubens), 이세 리아깍지벌레 (Icerya purchasi) 등; 방패벌레류 (Tingidae); 나무이류 (Psyllidae) 등);
- 인시류 (Lepidoptera) (예컨대, 다음을 포함함: 명나방류 (Pyralidae), 예컨대, 이화명나방 (Chilo suppressalis), 옐로우 라이스 나무좀 (Tryporyza incertulas), 혹명나방 (Cnaphalocrocis medinalis), 목화명나방 (Notarcha derogata), 화랑곡나방 (Plodia interpunctella), 조명나방 (Ostrinia furnacalis), 유럽조명나방 (Ostrinia nubilaris), 배추순나방 (Hellula undalis), 잔디포충나방 (Pediasia teterrellus) 등; 밤나방류 (Noctuidae), 예컨대, 담배거세미나방 (Spodoptera litura), 파밤나방 (Spodoptera exigua), 멸강나방 (Pseudaletia separata), 도둑나방 (Mamestra brassicae), 검거세미나방 (Agrotis ipsilon), 가두배추금날개밤나비 (Plusia nigrisigna), 토리코플루시아류 (Thoricoplusia spp.), 담배밤나방류 (Heliothis spp.), 담배나방류 (Helicoverpa spp.) 등; 흰나비류 (Pieridae), 예컨대, 배추흰나비 (Pieris rapae) 등; 잎말이나방류 (Tortricidae), 예컨대, 애모무늬잎말이나방류 (Adoxophyes spp.), 복숭아순나방 (Grapholita molesta), 콩나방 (Leguminivora glycinivorella), 팥나방 (Matsumuraeses azukivora), 사과애모무늬잎말이나방 (Adoxophyes orana fasciata), 차애모무늬잎말이나방 (Adoxophyes honmai), 차잎말이나방 (Homona magnanima), 검모무늬잎말이나방 (Archips fuscocupreanus), 코드린나방 (Cydia pomonella) 등; 가는나방류 (Gracillariidae), 예컨대, 동백가는나방 (Caloptilia theivora), 사과굴나방 (Phyllonorycter ringoniella) 등; 심식나방류 (Carposinidae), 예컨대, 복숭아심식나방 (Carposina niponensis) 등; 굴나방류 (Lyonetiidae), 예컨대, 은무늬굴나방류 (Lyonetia spp.) 등; 독나방류 (Lymantriidae), 예컨대, 매미나방류 (Lymantria spp.), 독나방류 (Euproctis spp.) 등; 집나방류 (Yponomeutidae), 예컨대, 배추좀 나방 (Plutella xylostella) 등; 뿔나방류 (Gelechiidae), 예컨대, 목화다래나방 (Pectinophora gossypiella), 감자뿔나방 (Phthorimaea operculella) 등; 불나방류 (Arctiidae), 예컨대, 미국흰불나방 (Hyphantria cunea) 등; 곡식좀나방류 (Tineidae), 예컨대, 옷좀나방 (Tinea translucens) 등);
- 총채벌레목 (Thysanoptera), 예컨대, 다음을 포함함: 꽃노랑총채벌레 (Frankliniella occidentalis), 오이총채벌레 (Thrips palmi), 볼록총채벌레 (Scirtothrips dorsalis), 파총채벌레 (Thrips tabaci), 대만총채벌레 (Frankliniella intonsa), 담배총채벌레 (Frankliniella fusca) 등;
- 쌍시류 (Diptera) (예컨대, 다음을 포함함:
굴파리류 (Agromyzidae), 예컨대, 고자리파리 (Hylemya antiqua), 씨고자리파리 (Hylemya platura), 벼잎굴파리 (Agromyza oryzae), 벼애잎굴파리 (Hydrellia griseola), 벼노랑굴파리 (Chlorops oryzae), 아메리카잎굴파리 (Liriomyza trifolii) 등; 멜론파리 (Dacus cucurbitae), 지중해과실파리 (Ceratitis capitata) 등);
- 초시류 (Coleoptera), 예컨대, 점박이 무당벌레 (Epilachna vigintioctopunctata), 오이잎벌레 (Aulacophora femoralis), 벼룩잎벌레 (Phyllotreta striolata), 벼잎벌레 (Oulema oryzae), 벼뿌리바구미 (Echinocnemus squameus), 벼물바구미 (Lissorhoptrus oryzophilus), 목화바구미 (Anthonomus grandis), 팥바구미 (Callosobruchus chinensis), 헌팅 바구미 (Sphenophorus venatus), 왜콩풍뎅이 (Popillia japonica), 구리풍뎅이 (Anomala cuprea), 옥수수잎벌 레류 (Diabrotica spp.), 콜로라도감자잎벌레 (Leptinotarsa decemlineata), 방아벌레류 (Agriotes spp.), 궐련벌레 (Lasioderma serricorne) 등;
- 메뚜기류 (Orthoptera), 예컨대, 땅강아지 (Gryllotalpa africana), 벼메뚜기 (Oxya yezoensis), 벼메뚜기 (Oxya japonica) 등;
- 막시류 (Hymenoptera), 예컨대, 무잎벌 (Athalia rosae), 가위개미(일개미)류 (Acromyrmex spp.), 불개미류 (Solenopsis spp.) 등;
- 이목 (Phthiraptera), 예컨대, 소 이 (Damalinia bovis, Haematopinus eurysternus, Linognathus vituli), 돼지 이 (Haematopinus suis) 등;
- 진드기류 (Acarina) (예컨대, 다음을 포함함:
응애류 (Tetranychidae), 예컨대, 점박이응애 (Tetranychus urticae), 귤응애 (Panonychus citri), 나무응애 류 (Oligonychus spp.) 등; 혹응애류 (Eriophyidae), 예컨대, 귤녹응애 (Aculops pelekassi); 먼지응애류 (Tarsonemidae), 예컨대, 차먼지응애 (Polyphagotarsonemus latus); 지응애류 (Tenuipalpidae); 치레응애류 (tuckerellidae); 가루진드기류 (Acaridae), 예컨대, 긴털가루진드기 (Tyrophagus putrescentiae) 등; 먼지진드기 류 (Pyroglyphidae), 예컨대, 큰다리먼지진드기 (Dermatophagoides farinae), 세로무늬먼지진드기 (Dermatophagoides ptrenyssnus) 등; 발톱진드기류 (Cheyletidae), 예컨대, 짧은빗살발톱진드기 (Cheyletus eruditus), 발톱진드기 (Cheyletus malaccensis), 케일레투스 무레이 (Cheyletus moorei) 등; 옴진드기류 (scabies mites), 예컨대, 옴진드기 (Sarcoptes scabiei) 등;
선충류 (Nematodes), 예컨대, 소나무재선충 (Bursaphelenchus xylophilus), 뿌리혹선충 (Meloidogyne spp.) 벼잎선충 (Aphelenchoides besseyi), 딸기아선충 (Nothotylenchus acris), 개구충 (Ancylostoma caninum), 모두충 (Capillaria spp.), 개사상충 (Dirofilaria immitis), 가축에 기생하는 선충류 (roundworn) 등.
상기 작물은 해충에 의하여 피해를 입을 수 있는 모든 식물일 수 있다.
상기 작물에 적용하는 단계는 침지, 분사, 도포 등의 통상적인 모든 방법에 의하여 수행될 수 있다.
본 명세서에 기재된 유전자 서열 및 아미노산 서열은 기능성 동등성을 유지하는 한, 60% 이상, 65% 이상, 70% 이상, 75% 이상, 80% 이상, 85% 이상, 90% 이상, 92% 이상, 94% 이상, 96% 이상, 98% 이상, 또는 99% 이상의 서열 상동성을 갖는 경우를 포함하는 것으로 해석될 수 있다.
본 발명에서 제공된 재조합 균주를 이용하여 밀베마이신을 대량으로 생산할 수 있으며, 이에 의해 생산된 밀베마이신은 농업 및 동물의약품 분야에 폭넓게 활용될 수 있다.
도 1은 일 실시예에 따른 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 milA1 유전자의 구성을 나타낸 것이다.
도 2는 일 실시예에 따른 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 milA1 유전자를 확보하는 과정을 나타낸 것이다.
도 3은 pGEMB2-MA-DOWN-fu 벡터를 구성하는 염기서열을 나타낸 것이다.
도 4a 내지 도 4c는 일 실시예에 따른 pCR2.1-milA1-AC2 벡터를 제조하는 과정을 나타낸 것이다.
도 5는 일 실시예에 따른 pCR2.1-Adu-M1Age 벡터를 제조하는 과정을 나타낸 것이다.
도 6은 일 실시예에 따른 pCR2.1-Adu-milA1-Ac2 벡터를 제조하는 과정을 나타낸 것이다.
도 7a는 일 실시예에 따른 milA1 integration 벡터가 도입된 균주에서 교차를 통해 milA1유전자가 삽입되는 과정을 나타낸 모식도이다.
도 7b는 일 실시예에 따른 milA1 integration 벡터가 도입된 균주에서 milA1이 성공적으로 삽입되었는지를 확인한 결과이다.
도 8은 일 실시예에 따른 S. avermitilis DBM-01 균주의 배양액으로부터 생성된 물질을 분석한 HPLC 결과를 나타낸다.
도 9a 및 도 9b는 일 실시예에 따른 pCR2.1-A3Mmo7A 및 pCR2.1-A3Mmo7B 벡터를 제조하는 과정을 나타낸 것이다.
도 10a는 일 실시예에 따른 모듈7 교체 벡터가 도입된 균주에서 교차를 통해 모듈7 유전자가 삽입되는 과정을 나타낸 모식도이다.
도 10b는 일 실시예에 따른 모듈7 교체 벡터가 도입된 균주에서 모듈 7 이 성공적으로 삽입되었는지를 확인한 결과이다.
도 11a 내지 도 11c는 실시예에 따른 DBM-03-A와 DBM-03-B가 생성한 물질을 분석한 HPLC 분석 결과를 보여준다. 형성된 각 peak 중 retention time이 표기된 peak는 Milbemycin 표준품과 동일한 UV-흡광패턴 보여주는 것이다. 도 11a는 밀베마이신 표준품의 HPLC 분석결과, 도 11b는 S. avermitilis DBM-03-A가 생성한 물질의 HPLC 분석결과, 도 11c는 S. avermitilis DBM-03-B가 생성한 물질의 HPLC 분석결과를 각각 나타낸다.
도 12는 일 실시예에 따른 모듈7 치환 균주의 배양액으로부터 생성된 물질을 분석한 HPLC 결과 및 각 피크에서 생성된 물질을 나타낸다.
도 13은 aveA3 유전자와 milA3 유전자의 AT-DH 모듈간 영역의 코딩 유전자 부위 및 KR-ACP 모듈간 영역의 코딩 유전자 부위의 염기서열 분석 결과를 보여준다.
도 14는 일 실시예에 따른 milA3의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인까지의 영역을 코딩하는 유전자 부위를 포함하는 pD7K9milA3 벡터의 제작 과정을 모식적으로 보여준다.
도 15는 일 실시예에 따른 milA3 삽입 벡터 pD7K9milA3가 삽입된 스트렙토마이세스 아베미틸리스 균주의 제작 과정을 모식적으로 보여준다.
도 16은 일 실시예에 따라서 제작된 S. avermitilis DBM-03-C 균주의 생성 물질의 HPLC 분석 결과를 보여주는 그래프이다.
도 17a 내지 17f는 일 실시예에 따라서 제작된 S. avermitilis DBM-03-C 균주의 생성 물질의 UPLC-qTOF-HR-MS 분석 결과를 보여주는 그래프이다 (도 17a: 밀베마이신 A3; 도 17b: 밀베마이신 A4; 도 17c: 밀베마이신 D; 도 17d: 밀베마이신 alpha2; 도 17e: 밀베마이신 alpha4; 도 17f: 밀베마이신 G).
도 18a-18c는 아베멕틴과 밀베마이신의 분자구조 차이 및 이를 유발하는 유전자의 구성을 보여주는 그림이고, 도 18d는 밀베마이신 폴리케타이드 합성효소 유전자군 (gene cluster)과 아베멕틴 폴리케타이드 합성효소 유전자군을 비교하여 보여주는 그림이다.
도 2는 일 실시예에 따른 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 milA1 유전자를 확보하는 과정을 나타낸 것이다.
도 3은 pGEMB2-MA-DOWN-fu 벡터를 구성하는 염기서열을 나타낸 것이다.
도 4a 내지 도 4c는 일 실시예에 따른 pCR2.1-milA1-AC2 벡터를 제조하는 과정을 나타낸 것이다.
도 5는 일 실시예에 따른 pCR2.1-Adu-M1Age 벡터를 제조하는 과정을 나타낸 것이다.
도 6은 일 실시예에 따른 pCR2.1-Adu-milA1-Ac2 벡터를 제조하는 과정을 나타낸 것이다.
도 7a는 일 실시예에 따른 milA1 integration 벡터가 도입된 균주에서 교차를 통해 milA1유전자가 삽입되는 과정을 나타낸 모식도이다.
도 7b는 일 실시예에 따른 milA1 integration 벡터가 도입된 균주에서 milA1이 성공적으로 삽입되었는지를 확인한 결과이다.
도 8은 일 실시예에 따른 S. avermitilis DBM-01 균주의 배양액으로부터 생성된 물질을 분석한 HPLC 결과를 나타낸다.
도 9a 및 도 9b는 일 실시예에 따른 pCR2.1-A3Mmo7A 및 pCR2.1-A3Mmo7B 벡터를 제조하는 과정을 나타낸 것이다.
도 10a는 일 실시예에 따른 모듈7 교체 벡터가 도입된 균주에서 교차를 통해 모듈7 유전자가 삽입되는 과정을 나타낸 모식도이다.
도 10b는 일 실시예에 따른 모듈7 교체 벡터가 도입된 균주에서 모듈 7 이 성공적으로 삽입되었는지를 확인한 결과이다.
도 11a 내지 도 11c는 실시예에 따른 DBM-03-A와 DBM-03-B가 생성한 물질을 분석한 HPLC 분석 결과를 보여준다. 형성된 각 peak 중 retention time이 표기된 peak는 Milbemycin 표준품과 동일한 UV-흡광패턴 보여주는 것이다. 도 11a는 밀베마이신 표준품의 HPLC 분석결과, 도 11b는 S. avermitilis DBM-03-A가 생성한 물질의 HPLC 분석결과, 도 11c는 S. avermitilis DBM-03-B가 생성한 물질의 HPLC 분석결과를 각각 나타낸다.
도 12는 일 실시예에 따른 모듈7 치환 균주의 배양액으로부터 생성된 물질을 분석한 HPLC 결과 및 각 피크에서 생성된 물질을 나타낸다.
도 13은 aveA3 유전자와 milA3 유전자의 AT-DH 모듈간 영역의 코딩 유전자 부위 및 KR-ACP 모듈간 영역의 코딩 유전자 부위의 염기서열 분석 결과를 보여준다.
도 14는 일 실시예에 따른 milA3의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인까지의 영역을 코딩하는 유전자 부위를 포함하는 pD7K9milA3 벡터의 제작 과정을 모식적으로 보여준다.
도 15는 일 실시예에 따른 milA3 삽입 벡터 pD7K9milA3가 삽입된 스트렙토마이세스 아베미틸리스 균주의 제작 과정을 모식적으로 보여준다.
도 16은 일 실시예에 따라서 제작된 S. avermitilis DBM-03-C 균주의 생성 물질의 HPLC 분석 결과를 보여주는 그래프이다.
도 17a 내지 17f는 일 실시예에 따라서 제작된 S. avermitilis DBM-03-C 균주의 생성 물질의 UPLC-qTOF-HR-MS 분석 결과를 보여주는 그래프이다 (도 17a: 밀베마이신 A3; 도 17b: 밀베마이신 A4; 도 17c: 밀베마이신 D; 도 17d: 밀베마이신 alpha2; 도 17e: 밀베마이신 alpha4; 도 17f: 밀베마이신 G).
도 18a-18c는 아베멕틴과 밀베마이신의 분자구조 차이 및 이를 유발하는 유전자의 구성을 보여주는 그림이고, 도 18d는 밀베마이신 폴리케타이드 합성효소 유전자군 (gene cluster)과 아베멕틴 폴리케타이드 합성효소 유전자군을 비교하여 보여주는 그림이다.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명이 하기 실시예에 의해 한정되는 것은 아니다.
실시예
1.
milA1
삽입 벡터(
milA1
integration vector) 제작
밀베마이신 생산 균주인 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 milA1 유전자를 S. avermitilis SA-01의 aveA1에 치환하기 위한 벡터를 제조하였다. 상기 벡터는 milA1 유전자 전체를 포함하고, 통합을 위한 상동성을 가지는 aveA1 유전자의 upstream 및 downstream 지역이 milA1의 양쪽 말단에 붙여진 형태로 구성하였다. 이하, 벡터 제작 과정을 상세히 설명한다.
1-1.
milA1
유전자 확보
스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739) genomic DNA로부터 milA1 유전자를 확보하였다. 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 경우 밀베마이신 생합성 유전자들의 DNA 염기서열이 알려져 있지 않기 때문에, NCBI에 등록된 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis; Accession no. FJ952082) 또는 스트렙토마이세스 빙쳉젠시스(Streptomyces bingchenggensis; Accession no. CP002047.1)의 밀베마이신 생합성 유전자군의 염기서열을 기반으로 프라이머를 디자인하여 유전자를 증폭하여 이용하였다. 또한, 밀베마이신 생합성에 관여하는 폴리케타이드 생합성효소의 유전자는 매우 크고 높은 GC비율을 갖기 때문에 한 번에 증폭하는 것은 매우 어려운 일로 이를 극복하기 위해 유전자를 임의의 두 구역으로 나눠 증폭하였다 (도 1).
우선, milA1 유전자의 N-terminus쪽의 단편을 확보하기 위하여, 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 genomic DNA를 A1NT2-F 프라이머(GAACCGTGTGCGGAAGAA: 서열번호 1) 및 A1CT1-R 프라이머(ATCAGGTCGGTGAGACGGT: 서열번호 2)를 사용하여 PCR(95도에서 5분; 98도에서 20초, 60도에서 20초 및 68도에서 10분으로 30 사이클; 및 68도에서 10분)을 통해 증폭한 후 pCR2.1 벡터(TA 클로닝 벡터)에 클로닝하였다. 이 벡터를 pCR2.1-NM으로 명명하였다. 다음으로, milA1 유전자의 C-terminus쪽의 단편을 확보하기 위하여, 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 genomic DNA를 A1CT1-F 프라이머(GCAAGGTCGTCTTTGTGTGC: 서열번호 3) 및 A1CT2-R 프라이머(CCACCAACTCGATATGGGC: 서열번호 4)를 사용하여 PCR(95도에서 5분; 98도에서 20초, 60도에서 20초 및 68도에서 10분으로 30 사이클; 및 68도에서 10분)을 통해 증폭한 후 pCR2.1 벡터(TA 클로닝 벡터)에 클로닝하였다. 이 벡터를 pCR2.1-CT으로 명명하였다. 상기 제조한 pCR-NM과 pCR-CT 벡터를 각각 NcoI 제한효소로 처리하여 단편(NM fragment, CT fragment)을 확보한 후(도 1), 이들 단편을 연결(ligation)하여 하나의 벡터를 제조하였으며, 이를 pCR2.1-milA1으로 명명하였다(도 2; milA1 gene 서열 (서열번호: 70) 포함). 이 때, 연결된 단편의 ORF의 염기서열을 확인한 결과, 스트렙토마이세스 빙쳉젠시스(Streptomyces bingchenggensis)의 염기서열과 100% 일치하는 것을 확인하였다.
1-2. C-
terminus쪽
arm 제작
milA1의 모듈 2의 위치에 해당하는 일부 DNA sequence와, aveA1 docking domain으로 추정되는 부위, 및 aveA2 sequence를 포함하는 DNA를 인공적으로 합성하였고, 합성된 DNA는 pGEM vector에 도입하여 pGEMB2-MA_DOWN_fu를 제작하였다(도 3; 서열번호 35 내지 서열번호 37). 상기 제작된 pGEMB2-MA_DOWN_fu를 BamHI/HindIII 제한효소로 잘라 insert를 분리한 후, BamHI/HindIII로 자른 pCR2.1-CT에 연결하여 milA1의 C-terminal 부위가 aveA1의 C-terminal 부위로 교체된 pCR2.1-CT-Ac 벡터를 제작하였다.
pCR2.1-CT-Ac 벡터의 결합부위를 연장하기 위하여, S. avermitilis SA-01균주의 genomic DNA로부터 A2-404F 프라이머(GGCGCTGGCAATGGACC: 서열번호 5) 및 A2PH-1500R 프라이머(GATTACGCCAAGCTTGGCTGATTCTCCCGGGACG: 서열번호 6)를 이용하여 aveA2의 N-terminus region을 PCR 증폭하고, 이를 infusion ligation을 통해 pCR2.1-CT-AC의 3' 말단에 결합하여, pCR2.1-CT-Ac2 벡터를 제작하였다.
pCR2.1-CT-Ac2 벡터와 pCR2.1-NM 벡터를 각각 NdeI/XbaI 제한효소로 처리한 후 결합하여, milA1 유전자의 C-terminal부분이 aveA1의 것으로 치환된 milA1 유전자를 갖는 pCR2.1-milA1-Ac2 벡터를 제작하였다(도 4a 내지 도 4c 참조).
1-3. N-
terminus쪽
arm 제작
S. avermitilis SA-01 균주의 genomic DNA로부터 AveD2-F 프라이머(GATGACGGAAATCCCCTGG: 서열번호 7), AveD2-R 프라이머(AGAATGAAAGGAGCGCGA: 서열번호 8), MA1NU-F 프라이머(GAGGACACAGCTCAGTTGCCCAAAGCCCAGAAC: 서열번호 9), M1Age-R 프라이머(TGATCGAAGAGCCGGGAGT: 서열번호 10), PHR-M1Ag 프라이머(CCGGCTCTTCGATCAAGCTTGGCGTAATCATGG: 서열번호 11) 및 PXF-AD2 프라이머(GGGATTTCCGTCATCAAGCTTGGCGTAATCATGG: 서열번호 12)를 이용하여 N-terminus arm에 해당하는 3개의 단편을 각각 PCR 증폭하고, 이를 infusion ligation을 통해 결합하여 pCR2.1-Adu-M1Age 벡터를 제작하였다(도 5).
1-4.
milA1
유전자에 양쪽 arm이
결합된
형태의 vector 제작
pCR2.1-milA1-Ac2 벡터와 pCR2.1-Adu-M1Age 벡터를 XmnI 제한효소로 처리한 후 이를 infusion ligation을 통해 결합하여, pCR2.1-Adu-milA1-Ac2 벡터 (서열번호 79)를 제작하였다(도 6). 이를 아베멕틴 생산균주로 도입하기 위해 스트렙토마이시스-대장균 셔틀벡터인 pKC1139 벡터(M. Bierman et al., Gene, 116:43-49)와 pCR2.1-Adu-milA1-Ac2 벡터를 각각 XbaI, HindIII 제한효소로 처리한 후 연결(ligation)하여 pKC1139에 Adu-milA1-Ac2 구조가 들어가 있는 형태인, pKC-Adu-milA1-Ac2 벡터를 제작하였다.
실시예
2.
milA1
치환 균주 제작 및 생성물질 확인
milA1 integration 벡터를 S. avertmilis SA-01 균주 내로 도입시키고, 상동재조합이 일어나도록 유도하여 S. avertmilis SA-01 균주의 aveA1 유전자가 milA1 유전자로 치환된 형태의 균주를 확보하고, 배양을 통해 생성되는 신규물질을 확인하였다.
2-1. 접합법을 통한 형질전환
pKC-Adu-milA1-Ac2 벡터를 포함하고 있는 E. coli 균주(ET12567/pUZ8002 strain)를 chloramphenicol, kanamycin, apramycin을 넣은 LB 액체배지 3ml에 접종한 후 Shaking incubator에서 37, 200rpm 조건으로 18~24시간 배양하였다. 멸균된 250ml flask에 25ml LB broth를 위의 항생제와 함께 넣고, 배양액 250ul을 접종한 뒤 동일 조건으로 3시간 배양하였다다. OD가 0.4~0.5에 이르렀을 때, 배양을 종료하고 5000rpm 에서 5분간 원심분리하였고(4 조건), Pellet만 취하여 냉각된 LB broth 1ml에 풀어주고 세척하여 균체 현탁액을 확보하였다. 또한, S. avertmilis SA-01 균주를 germination 하기 위하여, ISP4 agar plate에 host로 사용할 방선균을 7일 이상 배양하고, Spore를 loop로 긁어서 2X TY 배지에 풀어주고, 50 heating block에 넣고 10분간 activation을 실시하였다. 이와 같이 준비된 E. coli 균주 500ul과 germination된 host 균주 S. avertmilis SA-01 500ul을 microtube에 넣고 inverting으로 섞고, 8000rpm 에서 2분간 원심분리하여 상층액을 제거하는 방법으로 접합(conjugation)을 유도하여, pKC-Adu-milA1-Ac2 벡터를 S. avertmilis SA-01 균주 내로 도입시켰다.
2-2.
교차에 의한 유전자 치환
상기와 같이 pKC-Adu-milA1-Ac2(Et) 벡터가 도입된 균주에서 1차 교차를 유도하여 벡터 전체가 균주의 염색체에 삽입되도록 하였다. 이를 위하여, apramycin이 25ug/ml 농도로 첨가된 ISP-4 고체배지를 제조한 후, pKC-Adu-milA1-Ac2 벡터 도입에 의해 생성된 colony를 취한 후, 준비된 고체 배지에 streaking 하였다. 이후 고온에 민감한 pKC1139 벡터의 성질을 이용하여 벡터가 복제 되는 것을 막기 위하여 37에서 7일간 배양하였다.
2-3.
2차 교차
상기 1차 교차가 일어난 균주에서 aveA1 유전자를 milA1 유전자로 치환하기 위해 2차 교차를 유도하였다(도 7). 이를 위하여, apramycin이 25ug/ml 농도로 첨가된 ISP-4 고체배지와 항생제가 첨가 되지 않은 배지를 각각 제조하고, 1차 교차를 유도한 균주를 ISP-4 고체 배지에 도말하여 2차 교차를 유도한 후, apramycin 저항성 유무로 후보 균주를 선발하였다. 스크리닝을 통해 확보한 후보 균주의 genomic DNA를 추출하여, aveA1, milA1 일부지역을 확인할 수 있는 프라이머(표 1)를 사용하여 유전자의 교체가 정확하게 이루어졌는지 확인하였다. 이와 같이 최종 균주를 선발하고 S. avermitilis DBM-01로 명명하였으며, KCTC(Korean Colletion for Type Cultures, 한국생명공학연구원 미생물자원센터)에 기탁하여 2015년 8월 31일자로 기탁번호 KCTC12889BP 를 부여받았다.
| Primer Name | Sequence | 용도 |
| aveFi-F | GGCAACACCAACCGTGAGAT(서열번호 13) | N-terminus fusion 확인 |
| milA1i-R | GATGACCGAGGAGTGGAACG(서열번호 14) | |
| milA1i-F | GTACTGGACCTGTTCGAGCG(서열번호 15) | C-terminus fusion 확인 |
| aveA2i-R2 | ATGAATGCGGTCTTTCCTGC(서열번호 16) | |
| aveA1C-F | GGACTCGCTGATTCGGACAA(서열번호 17) | aveA1 일부지역 확인 |
| aveA1C-R | GGAGCAGATGAATGCCCTCT(서열번호 18) | |
| milA1C-F | CCTGTGGGAGTTCATCACCG(서열번호 19) | milA1 일부지역 확인 |
| milA1C-R | CGAGATTGTGGTGGACGAGG(서열번호 20) |
2-4.
S.
avermitilis
DBM-01의 생성물질 확인
S. avermitilis DBM-01 균주를 배양하기 위한 종균 배양용 배지와 생산 배양용 배지를 각각 제조하였다. 종균 배지를 제조하기 위하여, soluble starch 30g/L(Junsei, Japan), yeast extract 15g/L(Duchefa, 네덜란드), KH2PO4 0.4g/L(Junsei)의 농도가 되도록 적정량의 DW에 혼합한 후, pH를 7.2로 맞추고 121, 15분간 고압살균을 수행하였다. 충분히 식은 후, 별살한 Corn steep liquor(Sigma)를 5g/L의 농도가 되도록 첨가하였다. 또한, 생산 배지를 제조하기 위하여, soluble starch 80g/L, soybean meal 10g/L(Sigma), skim milk 15g/L(Difco), KH2PO4 0.5 g/L의 농도가 되도록 적정량의 DW에 혼합한 후, pH를 7.2로 맞추고, 121, 15분간 고압살균을 실시하였다. 생산 배양에 접종할 균주의 양을 충분히 만들기 위해, 상기 제조한 종균 배지 25ml을 멸균된 250ml baffled flask에 넣고, 균주 mycelium을 agar plate로부터 1loop씩 떠서 종균 배지에 접종하였다. 230 rpm 및 28도 조건하에서 48시간 동안 배양하였다. 또한, 생산배지 25ml을 멸균된 250ml non-baffled flask 넣고 종균 배양액 1.25ml을 접종한 후, 230 rpm 및 28도 조건하에서 10일 동안 배양하였다. 생산 배양액으로부터 생성된 물질을 추출하기 위하여, 유기용매(acetonitrile : MeOH = 1 : 1 혼합 용액) 1.5ml과 배양액 0.5ml을 섞고 충분히 vortexing하였다. 5분간 sonication 한 후, 1시간 동안 교반하였다. 12000rpm에서 5분간 원심분리한 후 상등액만 취하여 분석하였다.
추출한 시료를 아래의 조건δ에서 HPLC/UVD 분석을 통해 분석하였다.
- Mobile phase : Acetonitrile/Water(v/v)= 40/60, v/v(5min), 90/10, v/v(30min), 90/10, v/v(10min)
- Flow rate : 1 ml/min
- wavelength : 245 nm
- Run time : 45 min
또한, DBM-01을 배양하여 나온 생성 물질 중 신규한 물질 4종(도 8에서 IV1, IV2, IV3, IV5로 표시함)에 대해 NMR 분석을 실시하여 구조를 분석한 결과는 다음과 같다:
600 MHz in CD3OD(the internal references residual solvent protons and carbons; δH 4.78, δC 49.15)
IV1: δH: 3.16(m, H-2), 5.35(d, J = 1.4, H-3), 1.75(s, 3H, H-4a), 4.16(m, H-5), 3.70(m, H-6), 4.55(m, H-8a), 5.74(dd, J = 1.9 & 11.3, H-9), 5.81(ddd, J = 3.4 & 11.4 & 14.2, H-10), 5.65(ddd, J = 1.9 & 9.7 & 14.8, H-11), 2.59(m, H-12), 1.09(m, H-12a), 3.93(m, H-13), 1.48(s, 3H, H-14a), 5.15(m, H-15), 2.24(m, H-16), 3.63(m, H-17), 0.75(m, H-18), 1.86(m, H-18), 4.98(m, H-19), 1.14(m, H-20), 2.11(m, H-20), 1.24(m, H-22), 1.44(m, H-23), 1.52(m, H-23), 1.18(m, H-24), 0.77(d, J = 6.6, 3H, H-24a), 3.25(m, H-25), 1.08(m, H-26), 4.76(m, H-1'), 2.23(m, H-2'), 3.46(m, H-3'), 3.34(s, 3H, 3'-OMe), 3.12(m, H-4'), 3.76(m, H-5'), 1.17(m, H-6'), 5.26(d, J = 3.2, H-1"), 1.57(m, H-2"), 3.36(H-3"), 3.40(s, 3H, 3"-OMe), 3.64(m, H-4"), 2.97(m, H-5"), 1.15(m H-6") δC: 173.6(C-1), 47.1(C-2), 120.3(C-3), 137.2(C-4), 19.9(C-4a), 69.1(C-5), 82.3(C-6), 82.2(C-7), 141.9(C-8), 68.7(C-8a), 121.8(C-9), 126.7(C-10), 138.4(C-11), 41.2(C-12), 21.0(C-12a), 82.7(C-13), 136.3(C-14), 15.4(C-14a), 120.1(C-15), 35.2(C-16), 69.1(C-17), 38.1(C-18), 69.9(C-19), 42.7(C-20), 99.1(C-21), 30.7(C-22), 36.9(C-23), 38.0(C-24), 18.2(C-24a), 72.7(C-25), 19.8(C-26), 96.3(C-1'), 36.2(C-2'), 79.7(C-3'), 57.0(3'-OMe), 82.3(C-4'), 69.9(C-5'), 19.1(C-6'), 99.9(C-1"), 37.3(C-2"), 79.5(C-3"), 57.6(3"-OMe), 69.2(C-4"), 77.1(C-5"), 18.3(C-6"). HRESQTOF m/z 855.4507 [M+Na]+(cald for C45H68O14Na)
IV2: δH: 3.16(m, H-2), 5.36(d, J = 1.5, H-3), 1.75(s, 3H, H-4a), 4.17(m, H-5), 3.71(m, H-6), 4.55(m, H-8a), 5.75(dt, J = 2.2 & 11.3, H-9), 5.82(dd, J = 11.2 & 14.9, H-10), 5.65(dd, J = 9.8 & 14.9, H-11), 2.59(m, H-12), 1.10(d, J = 7.0, H-12a), 3.92(m, H-13), 1.48(s, 3H, H-14a), 5.13(m, H-15), 2.44(m, H-16), 3.62(m, H-17), 0.77(m, H-18), 1.85(m, H-18), 5.00(m, H-19), 1.16(m, H-20), 2.13(m, H-20), 1.26(m, H-22), 1.66(m, H-22), 1.15(m, H-23), 1.43(m, H-23), 1.24(m, H-24), 0.76(d, J = 6.5, 3H, H-24a), 3.05(m, H-25), 1.29(m, H-26), 0.93(t, J = 7.3, H-27), 4.75(m, H-1'), 2.24(m, H-2'), 3.56(m, H-3'), 3.32(s, 3H, 3'-OMe), 3.12(m, H-4'), 3.65(m, H-5'), 1.18(d, J = 6.2, H-6'), 5.26(d, J = 3.2, H-1"), 1.56(m, H-2"), 3.35(H-3"), 3.35(s, 3H, 3"-OMe), 3.64(m, H-4"), 2.94(m, H-5"), 1.15(d, J = 6.2, H-6") δC: 173.6(C-1), 47.1(C-2), 120.1(C-3), 137.2(C-4), 19.9(C-4a), 69.1(C-5), 82.2(C-6), 82.0(C-7), 142.0(C-8), 68.6(C-8a), 121.8(C-9), 126.7(C-10), 138.4(C-11), 41.2(C-12), 20.9(C-12a), 83.2(C-13), 136.5(C-14), 15.3(C-14a), 120.3(C-15), 35.1(C-16), 69.1(C-17), 38.1(C-18), 70.1(C-19), 42.9(C-20), 99.0(C-21), 30.9(C-22), 29.2(C-23), 35.6(C-24), 18.2(C-24a), 77.5(C-25), 26.9(C-26), 10.6(C-27), 96.4(C-1'), 36.2(C-2'), 80.9(C-3'), 56.9(3'-OMe), 82.2(C-4'), 70.1(C-5'), 19.1(C-6'), 99.9(C-1"), 36.8(C-2"), 79.4(C-3"), 57.6(3"-OMe), 70.2(C-4"), 77.5(C-5"), 18.2(C-6"). HRESQTOF m/z 869.4663 [M+Na]+(cald for C46H70O14Na)
IV3: δH: 3.16(m, H-2), 5.42(d, J = 1.4, H-3), 1.74(s, 3H, H-4a), 3.92(m, H-5), 3.41(s, 5-OMe), 3.90(m, H-6), 4.58(m, H-8a), 5.77(dd, J = 1.9 & 11.3, H-9), 5.85(ddd, J = 3.4 & 11.4 & 14.2, H-10), 5.68(ddd, J = 1.9 & 9.7 & 14.8, H-11), 2.59(m, H-12), 1.09(m, H-12a), 3.90(m, H-13), 1.48(s, 3H, H-14a), 5.17(m, H-15), 2.24(m, H-16), 3.63(m, H-17), 0.75(m, H-18), 1.86(m, H-18), 5.00(m, H-19), 1.14(m, H-20), 2.11(m, H-20), 1.24(m, H-22), 1.44(m, H-23), 1.52(m, H-23), 1.18(m, H-24), 0.77(d, J = 6.6, 3H, H-24a), 3.25(m, H-25), 1.08(m, H-26), 4.76(m, H-1'), 2.23(m, H-2'), 3.46(m, H-3'), 3.34(s, 3H, 3'-OMe), 3.13(m, H-4'), 3.81(m, H-5'), 1.17(m, H-6'), 5.28(d, J = 3.2, H-1"), 1.57(m, H-2"), 3.36(H-3"), 3.40(s, 3H, 3"-OMe), 3.64(m, H-4"), 2.97(m, H-5"), 1.15(m H-6") δC: 173.4(C-1), 47.0(C-2), 121.4(C-3), 135.4(C-4), 20.0(C-4a), 78.56(C-5), 57.8(5-OMe), 80.9(C-6), 82.2(C-7), 141.5(C-8), 68.7(C-8a), 121.9(C-9), 126.7(C-10), 138.5(C-11), 41.2(C-12), 21.0(C-12a), 82.7(C-13), 136.3(C-14), 15.4(C-14a), 120.1(C-15), 35.2(C-16), 69.1(C-17), 38.1(C-18), 69.9(C-19), 42.7(C-20), 99.1(C-21), 30.7(C-22), 36.9(C-23), 38.0(C-24), 18.2(C-24a), 72.7(C-25), 19.8(C-26), 96.3(C-1'), 36.2(C-2'), 79.7(C-3'), 57.0(3'-OMe), 82.3(C-4'), 69.9(C-5'), 19.1(C-6'), 99.9(C-1"), 37.3(C-2"), 79.5(C-3"), 57.6(3"-OMe), 69.2(C-4"), 77.1(C-5"), 18.3(C-6"). HRESQTOF m/z 869.4662 [M+Na]+(cald for C46H70O14Na)
IV5: δH: 3.16(m, H-2), 5.41(d, J = 1.5, H-3), 1.74(s, 3H, H-4a), 3.93(m, H-5), 3.41(s, 5-OMe), 3.91(m, H-6), 4.57(m, H-8a), 5.76(dt, J = 2.2 & 11.3, H-9), 5.82(dd, J = 11.2 & 14.9, H-10), 5.66(dd, J = 9.8 & 14.9, H-11), 2.59(m, H-12), 1.11(d, J = 7.0, H-12a), 3.92(m, H-13), 1.47(s, 3H, H-14a), 5.13(m, H-15), 2.43(m, H-16), 3.62(m, H-17), 0.78(m, H-18), 1.85(m, H-18), 5.00(m, H-19), 1.15(m, H-20), 2.13(m, H-20), 1.25(m, H-22), 1.66(m, H-22), 1.14(m, H-23), 1.43(m, H-23), 1.24(m, H-24), 0.76(d, J = 6.5, 3H, H-24a), 3.04(m, H-25), 1.29(m, H-26), 0.93(t, J = 7.3, H-27), 4.75(m, H-1'), 2.24(m, H-2'), 3.56(m, H-3'), 3.32(s, 3H, 3'-OMe), 3.12(m, H-4'), 3.65(m, H-5'), 1.18(d, J = 6.2, H-6'), 5.26(d, J = 3.2, H-1"), 1.56(m, H-2"), 3.35(H-3"), 3.35(s, 3H, 3"-OMe), 3.64(m, H-4"), 2.94(m, H-5"), 1.15(d, J = 6.2, H-6") δC: 173.5(C-1), 47.1(C-2), 121.5(C-3), 135.3(C-4), 20.2(C-4a), 78.4(C-5), 57.7(5-OMe), 80.8(C-6), 82.1(C-7), 141.4(C-8), 68.8(C-8a), 121.9(C-9), 126.8(C-10), 138.3(C-11), 41.2(C-12), 21.0(C-12a), 83.3(C-13), 136.4(C-14), 15.3(C-14a), 120.4(C-15), 35.2(C-16), 69.1(C-17), 38.0(C-18), 70.2(C-19), 42.7(C-20), 99.1(C-21), 30.7(C-22), 29.2(C-23), 35.6(C-24), 18.2(C-24a), 77.5(C-25), 26.8(C-26), 10.6(C-27), 96.4(C-1'), 36.2(C-2'), 80.9(C-3'), 56.9(3'-OMe), 82.2(C-4'), 70.1(C-5'), 19.1(C-6'), 99.9(C-1"), 36.8(C-2"), 79.4(C-3"), 57.6(3"-OMe), 70.2(C-4"), 77.5(C-5"), 18.2(C-6"). HRESQTOF m/z 883.4823 [M+Na]+(cald for C47H72O14Na)
Ivermectin 유도체 1(IV1)의 구조는 화학식 1과 같이 결정하였으며, HMBC correlation을 화살표로 표시하였다.
[화학식 1]
22,23-dihydro-25-methyl-avermectin
Ivermectin 유도체 2(IV2)의 구조는 화학식 2와 같이 결정하였으며, HMBC correlation을 화살표로 표시하였다.
[화학식 2]
22,23-dihydro-25-ethyl-avermectin
Ivermectin 유도체 3(IV3)의 구조는 화학식 3과 같이 결정하였으며, HMBC correlation을 화살표로 표시하였다.
[화학식 3]
5-methoxy-22,23-dihydro-25-methyl-avermectin
Ivermectin 유도체 5(IV5)의 구조는 화학식 4와 같이 결정하였으며, HMBC correlation을 화살표로 표시하였다.
[화학식 4]
5-methoxy-22,23-dihydro-25-ethyl-avermectin
결과적으로, 상기 물질의 분석을 통해 avermectin 생산 균주인 S. avermitilis SA-01의 avermectin 생산 유전자군의 첫번째 유전자인 aveA1을 S. hygroscopicus subsp. aureolacrimosus의 밀베마이신 생산 유전자군의 첫번째 유전자인 milA1으로 교체하였을 때, 숙주 균주와 다른 형태의 물질 생산 패턴을 나타냄을 확인할 수 있었으며, milA1이 삽입된 균주는 각각 22,23-dihydro-25-methyl-avermectin (CAS No. 137330-81-5; Ref: DE4031039), 22,23-dihydro-25-ethyl-avermectin (CAS No. 71972-13-9;Ref: FR2387231), 22,23-dihydro-25-isopropyl-avermectin(즉, Ivermectin B1b, CAS NO. 70209-81-3; Ref. US4199569), 5-methoxy-22,23-dihydro-25-methyl-avermectin, 5-methoxy-22,23-dihydro-25-ethyl-avermectin, 및 5-methoxy-22,23-dihydro-25-isopropyl-avermectin (즉, Ivermectin A1b; Ref. US4199569)을 생성함을 알 수 있었다.
이들 중, 5-methoxy-22,23-dihydro-25-methyl-avermectin (화학식 3; IV3) 및 5-methoxy-22,23-dihydro-25-ethyl-avermectin (화학식 4; IV5)의 경우, 물질 구조를 이용한 SciFinder 검색에서 일치하는 구조를 찾지 못하였는 바, 신규 화합물인 것으로 판단되었다.
2-5.
S.
avermitilis
DBM-01 생성물질인 IV3와 IV5의 생물학적 활성
상기 실시예 2-4에서 신규 화합물로 확인된 IV3 (화학식 3) 및 IV4 (화학식 4)의 생물학적 활성을 평가하여, 상기 화합물류가 해충의 유충 및 성충, 특히 점박이응애와 배추좀나방의 해충 구제 (살충)에 활성효과가 있음을 확인하였다.
상기 화합물류의 살충 활성을 측정하기 위하여 다음과 같은 실험을 수행하였다. 상기 시험 화합물 IV3 및 IV4을 각각 용매인 acetone에 용해시키고 농도가 100ppm이 되도록 용해시켜 stock를 만든 후, 각 시험에서의 처리 농도 (표 2 및 표 3 참조)가 되도록 희석하여 사용하였다. 점박이 응애의 경우, 성충을 대상으로 하였으며, 상기 준비된 희석액을 배추 (Φ5cm/disk)에 1ml을 spray 처리하고, 1 내지 2일 후 생충수를 측정하였다. 이 경우, 대조약제는 아바멕틴(Sigma)을 사용하였다. 배추좀나방의 경우, 2령충을 대상으로 하였으며, 배추(Φ5cm/disk) 절편에 침지 처리한 후 시험충을 절편당 10마리 접종하여 1 내지 2일 후 생충수를 측정 하였다. 이 경우, 대조약제는 에바멕틴(Sigma)를 사용하였다.
상기 얻어진 결과를 아래의 표 그 결과를 표 2 및 표 3에 각각 나타내었다.
| 물질 | 처리 농도 | 처리기간 | |
| 1일차 | 2일차 | ||
| IV3 | 0.6 ppm | 100.0 | 100.0 |
| IV5 | 76.7 | 86.7 | |
| abamectin | 100.0 | 100.0 | |
| 물질 | 처리 농도 | 처리기간 (2일차) |
| IV3 | 0.06 ppm | 100 |
| IV5 | 100 | |
| Abamectin | 96.7 |
상기 표 3 및 표 4에 나타난 바와 같이, 화합물 IV3 및 IV4는 대조약물과 비교하여 동등 이상의 해충의 유충 및 성충에 대한 방제 활성을 나타냄을 확인할 수 있다.
실시예
3. 모듈 7 교체 벡터 제작
스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 milA3 유전자에 존재하는 7번 모듈을 S. avermitilis DBM-01의 물질 생산 유전자군의 7번 모듈과 치환하기 위한 벡터를 제조하였다. 상기 벡터는 7번 모듈 전체 또는 일부를 포함하고, 통합을 위한 상동성을 가지는 aveA3, aveA4 유전자의 지역이 양쪽 말단에 붙여진 형태로 구성하였다. 이하, 벡터 제작 과정을 상세히 설명한다.
3-1.
스트렙토마이세스
밀베마이시니쿠스
(S
.
milbemycinicus
NRRL5739
) genomic DNA로부터
모듈7
유전자 확보
모듈 7 전체와 모듈 7 중 KS(Ketosynthase) 도메인만 숙주 균주인 S. avermitilis DBM-01의 것을 사용한 두 가지 형태로 모듈의 치환을 계획하고 이에 따른 유전자를 확보하고자 하였다. 앞서 milA의 클로닝 경우와 마찬가지로 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 모듈 7의 염기서열이 알려져 있지 않기 때문에, NCBI에 등록된 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis) 또는 스트렙토마이세스 빙쳉젠시스(Streptomyces bingchenggensis)의 milA3의 염기서열을 활용하여 프라이머를 디자인하였다.
스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739) genomic DNA 로부터 모듈7에 해당하는 부위를 두 가지 형태로(A 및 B)로 클로닝하기 위하여, MA3-6503R 프라이머(TGCCAATGATCGCGATGGCCTCATC: 서열번호 21), MA3-104Fin(A) 프라이머(GGGTCAGTGGCACGAACGCCCATGTGATCCTCG: 서열번호 22) 및 MA3-1368Ffu(B) 프라이머(GGAGGCGTCCAGCGAACCCATCGCCATCATC: 서열번호 23)를 사용하여 PCR(95도에서 5분; 98도에서 20초, 60도에서 20초 및 68도에서 6분으로 30 사이클; 및 68도에서 10분)을 통해 증폭한 후 pCR2.1 벡터(TA 클로닝 벡터)에 클로닝하였다. 이 벡터를 pCR2.1-Mo7A, pCR2.1-Mo7B로 명명하였다 (도 9a 및 도 9b).
3-2.
양쪽 arm 제작
S. avermitilis DBM-01 균주의 모듈7 지역에 교차를 일으키기 위한 결합부위를 제작하였다. S. avermitilis DBM-01 균주의 genomic DNA를 아래 프라이머를 사용하여 증폭하여 양쪽 arm에 해당하는 부위의 fragment를 확보한 후, pCR2.1 벡터(TA 클로닝 벡터)에 각각 클로닝하였다.
AA3-8600RH : aaaaagcttAACGGTGTGTGGAGCGTCAG (서열번호 24)
AA3-5597FSp : aaaactagTCGCGATCATTGGCATG (서열번호 25)
AA3-111RSp : aaaactagtGCTGGACGCCTCCATGGC (서열번호 26)
AA3-1369RSp : aaaactagTCGTGCCACTGACCCCGAAC (서열번호 27)
AA4-2300FNs : aaatctagaGCTCCGTGCAGACCGAGAAC (서열번호 28)
벡터를 회수하여 NsiI/SpeI, SpeI/HindIII 제한효소로 각각 처리 한 후 ligation 하여 두 fragment를 붙인 형태로 양쪽 arm이 벡터 하나에 존재하도록 제작하였다. 이 벡터를 pCR2.1-armA, pCR2.1-armB 로 명명하였다(도 9).
확립한 pCR2.1-armA, pCR2.1-armB 벡터의 양쪽 arm 사이를 SpeI 제한효소로 자르고, pCR2.1-mo7A, pCR2.1-mo7B 벡터를 EcoRI 으로 잘라서 insert를 수득하였다. 이 두 단편을 각각 infusion ligation을 통해 결합하여 7번 모듈 교체 벡터의 구조를 완성하였다. 이 벡터를 각각 pCR2.1-A3Mmo7A, pCR2.1-A3Mmo7B로 명명하였다(도9)
pKC1139 벡터와 pCR2.1-A3Mmo7A, pCR2.1-A3Mmo7B 벡터를 각각 XbaI, HindIII 제한효소로 처리하고, pKC1139 벡터 fragment와 pCR2.1-A3Mmo7A, pCR2.1-A3Mmo7B 벡터의 insert fragment를 접합하여 pKC1139에 A3Mmo7A 또는 A3Mmo7B 구조가 들어가 있는 형태를 제작하였다. 이를 각각 pKC-A3Mmo7A, pKC-A3Mmo7B 벡터로 명명하였다 (도 9).
실시예
4. 모듈 7 교체 균주 제작 및 생성물질 확인
모듈7 교체 벡터를 DBM-01 균주 내로 도입시키고, 상동 재조합이 일어나도록 유도하여 S. avermitilis DBM-01 균주의 모듈7이 milA3의 모듈7로 치환된 형태의 균주를 확보하고, 배양을 통해 생성되는 신규물질을 확인하였다.
4-1.
PEG-mediated protoplast transformation을 통한 벡터 도입
Lysozyme을 처리하여 protoplast를 제조하고, pKC-A3Mmo7A(110), pKC-A3Mmo7B(110) 벡터를 섞고 PEG1000를 처리하여, PEG-mediated protoplast transformation을 통해 상기 벡터를 DBM-01 균주 내로 도입하였다. 24시간 배양한 후 항생제(apramycin)를 처리하였고, 위 벡터의 apramycin 저항성 유전자인 aac 유전자를 확인 할 수 있는 aac3iv-F 프라이머(GGTTCATGTGCAGCTCCATC: 서열번호 29) 및 aac3iv-R 프라이머(CGTCGCGGTGAGTTCAGG: 서열번호 30)를 사용하여 도입이 이루어졌음을 확인하였다.
4-2.
S.
avermitilis
DBM-01 균주로 모듈 7의 삽입
상기와 같이 모듈7 교체 벡터의 도입이 이루어진 균주에 1차 교차를 유도하여 벡터 전체가 S. avermitilis DBM-01 균주의 genomic DNA에 삽입되도록 하였다(도 10). 이를 위하여, apramycin이 25μg/ml 농도로 첨가된 ISP-4 고체배지를 제조한 후, 벡터 도입에 의해 생성된 colony를 취한 후, 준비된 고체 배지에 streaking 하였다. 이후 고온에 민감한 pKC1139 벡터의 성질을 이용하여 벡터가 복제 되는 것을 막기 위하여 37에서 7일간 배양하였다.
상기 1차 교차가 일어난 균주에서 aveA3의 모듈7을 milA3 모듈7로 치환하기 위해 2차 교차를 유도하였다(도 10). 이를 위하여, apramycin이 25ug/ml 농도로 첨가된 ISP-4 고체배지와 항생제가 첨가 되지 않은 배지를 각각 제조하고, 1차 교차를 유도한 균주를 ISP-4 고체 배지에 도말하여 2차 교차를 유도한 후, apramycin 저항성 유무로 후보 균주를 선발하였다. 스크리닝을 통해 확보한 후보 균주의 genomic DNA를 추출하여, aveA3 내의 모듈7, 및 milA3 내의 모듈7 일부지역을 확인할 수 있는 프라이머(표 4)를 사용하여 유전자의 교체가 정확하게 이루어졌는지 확인하였다 (도 10). 이를 통해 확보된 균주 중 A3Mmo7A가 삽입된 균주를 S. avermitilis DBM-03-A, A3Mmo7B가 삽입된 균주를 S. avermitilis DBM-03-B로 명명하였다. S. avermitilis DBM-03-A 균주는 2015년 8월 31일자로 기탁번호 KCTC12890BP 를 부여받았고, S. avermitilis DBM-03-B 균주는 2015년 8월 31일자로 기탁번호 KCTC12891BP 를 부여받았다.
| Primer Name | Sequence | 용도 |
| Amo7i-F | GTTGTCGGTGTTGCTTCCG (서열번호 31) | aveA3의 모듈7 확인 |
| Amo7i-R | CACATCACACGCAACAATCC (서열번호 32) | |
| Mmo7i-F | ACATCAGCGGCCTCTACGA (서열번호 33) | milA3의 모듈7 확인 |
| Mmo7i-R | CCAGGTCCATCACTCCCTC (서열번호 34) |
4-4.
모듈7
치환 균주의 생성물질 확인
모듈7 치환 균주를 배양하기 위한 종균 배양용 배지와 생산 배양용 배지를 각각 제조하였다. 종균 배지를 제조하기 위하여, soluble starch 30g/L, yeast extract 15g/L, KH2PO4 0.4g/L의 농도가 되도록 적정량의 DW에 혼합하여 교반기로 충분히 섞어주고, 충분히 식은 후, 별살한 Corn steep liquor 을 5g/L의 농도가 되도록 첨가하였다. 또한, 생산 배지를 제조하기 위하여, soluble starch 80g/L, soybean meal 10g/L, skim milk 15g/L, KH2PO4 0.5g/L의 농도가 되도록 적정량의 DW에 혼합하여 교반기로 충분히 섞어주고, pH를 7.2로 맞춘 후, 121, 15분간 고압살균을 실시하였다. 생산 배양에 접종할 균주의 양을 충분히 만들기 위해, 상기 제조한 종균 배지 25ml을 멸균된 250ml baffled flask에 넣고, 균주 mycelium을 agar plate로부터 1loop씩 떠서 종균 배지에 접종하였다. 230 rpm 및 28도 조건하에서 48시간 동안 배양하였다. 또한, 생산배지 25ml을 멸균된 250ml non-baffled flask 넣고 종균 배양액 1.25ml을 접종한 후, 230 rpm 및 28도 조건하에서 10일 동안 배양하였다. 생산 배양액으로부터 생성된 물질을 추출하기 위하여, 유기용매(acetonitrile : MeOH = 1 : 1 혼합 용액) 1.5ml과 배양액 0.5ml을 섞고 충분히 vortexing하였다. 5분간 sonication 한 후, 1시간 동안 교반하였다. 12000rpm에서 5분간 원심분리한 후 상등액만 취하여 분석하였다.
추출한 시료를 아래의 조건에서 HPLC/UVD 분석을 통해 분석하였다.
- Mobile phase : Acetonitrile/Water(v/v)= 40/60 v/v, 90/10 v/v, 90/10 v/v - Flow rate : 0.7 ml/min
- wavelength : 245 nm
- Run time : 40 min
그 결과, 밀베마이신 표준품의 HPLC 결과에서 나온 milbemycin A3와 A4의 peak (도 11a 참조)가 S. avermitilis DBM-03-A (도 11b 참조), S. avermitilis DBM-03-B 배양액 (도 11c 참조)의 HPLC분석 결과에서도 형성됨을 확인하였다(도 11a 내지 도 11c 참조).
또한, 모듈 7 치환 균주를 배양하여 수득한 생성물 중, HPLC로 확인 한 milbemycin A3, milbemycin A4 이외의 생성물질에 대한 분석을 실시하였다. 그 결과, HPLC 상에 보인 major peak에 대한 mass 분석을 실시하여 분자량을 근거로 예측했을 때, milbemycin α2, milbemycin α4, milbemycin G 가 생성 되었음을 확인할 수 있었다(도 12).
실시예
5.
aveA3의
변이를 위한
milA3
삽입 벡터(
milA3
integration vector) 제작
아베멕틴과 밀베마이신의 AT-DH 모듈간 영역 및 KR-ACP 모듈간 영역의 염기서열 분석을 통해 공통적인 amino acid를 코딩하는 지점을 치환이 일어나는 fusion site로 결정하고, amino acid의 서열에 변화를 주지 않는 범위 내에서 제한효소 site를 도입하였다. 상기 결과를 도 13에 나타내었으며, 하기의 유전자 변형을 위한 재조합 벡터의 제조의 제작시에 이를 참조하였다.
상기 실시예 2-3에서 제작된 스트렙토마이세스 아베미틸리스 DBM-01의 aveA3 유전자의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인을 포함하는 영역의 코딩 유전자 부위를 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 물질 생산 유전자군 milA3 유전자의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인 포함 영역의 코딩 유전자 부위로 치환하기 위한 벡터를 제작하였다. 이 벡터는 milA3의 일부를 포함하고 균주 내 재조합을 위한 상동성을 지니는 aveE 및 aveA3 유전자의 영역이 양쪽 말단에 연결된 형태로 구성하였다.
5-1.
MilA3
의 모듈 7의
DH
도메인부터 모듈 9의 KR 도메인까지의 코딩 유전자 부위의 확보
밀베마이신 생산 균주인 스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)로부터의 milA3 모듈 7 DH~모듈 9 KR 영역까지의 코딩 유전자의 확보를 위해 NCBI에 등록된 또 다른 밀베마이신 생산 균주 스트렙토마이세스 빙쳉젠시스 (Streptomyces binchenggensis)의 밀베마이신 폴리케타이드 합성효소의 염기서열을 활용하여, 하기의 표 5와 같은 염기서열로 프라이머를 디자인하였다 (제작하고자 하는 DNA fragment가 매우 거대하여, 3개의 구역(fragments)로 나누어 PCR을 수행한 후, 하기의 실시예 5-3의 과정을 통해 합성함).
| Primer | Sequence (5'→ 3') | 용도 |
| DK1F | GCGTCCGGGGTCGACATGGGATGCAGGCCTGAGCGGTTGATGCGGGCCCG (서열번호 38) | Fragment 1' |
| DK1R | ATGGGACTGCGGCTGCTTAAGACCTCACCCGTC(서열번호 39) | 합성 |
| DK2F | GCTAGCGGGGTGTGGTCG(서열번호 40) | Fragment 3 |
| DK2R | TTCGAACACGCGGGAATCGAC(서열번호 41) | |
| DK3F | GCATGCCGTGTCCACGCGCCC(서열번호 42) | Fragment 5-6' |
| DK3R | ACACCCACCCCCACAACCACCACCTAGATCTGCCCACCTACGCCTTCCA(서열번호 43) | 합성 |
상기 프라이머를 사용하여 얻어진 각 fragment를 PCR (1. 97 ℃ 3분, 2. 72℃ 1분, 3. 98 ℃ 10초, 4. 55 ℃ 15초, 5. 68 ℃ 1분 30초, 6. 스텝 3~ 스텝 5 에 대해31 사이클 반복 수행, 7. 72 ℃ 10분, 8. 4 ℃, 9. end)을 통해 증폭하였다. 각 fragment는 pGEMTeasy 벡터 (Promega)에 클로닝되어 DNA sequencing을 통해 염기서열을 확인하였다. 이와 같이 얻어진 벡터를 pYJ1950, pYJ1951, pYJ1952로 명명하였다. 이들 각 플라스미드의 insert는 도14의 fragment 1', 3과 5-6'에 해당하며, 각 fragment는 클로닝의 용이성을 위해 milA3 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인 영역에 존재하는 제한효소 위치를 참조하여 디자인 하였다.
5-2. Left arm 및 Right arm의 제작
스트렙토마이세스 아베미틸리스 DBM-01 균주의 모듈 7 DH~모듈 9 KR 영역에 상동재조합을 통한 교차를 일으키기 위해 각 벡터를 제작하였다. 스트렙토마이세스 아베미틸리스 DBM-01 균주의 genomic DNA를 주형으로 사용하고, DKLF/DKLR 및 DKRF/DKRR프라이머를 이용하여 (아래의 표 6 참조), milA3 유전자의 모듈 7 DH~모듈 9 KR 영역에 연결될 left 및 right flanking 영역 fragment를 PCR로 확보한 뒤, 각각 pGEMTeasy 벡터에 클로닝하였다. 이 때 각 fragment는 연결 될 milA3 영역과 상동성을 갖도록 디자인 되었고 이를 각각 pYJ1948 및 pYJ1949로 명명하였다. 각각 플라스미드의 insert는 도 14의 LA와 RA에 해당한다.
| Primer | Sequence (5'→3') | 용도 |
| DKLF | AACCCGAACGACCGAGCGCAGCGGCGGCCGCGAATTCATACGGCTACGAGGAT(서열번호 44) | Left arm (LA)의 제작 |
| DKLR | CTCGACCGCACCGACCGGGCCCGCATCAACCGCTCAGGCCTGCATCCCATGTCGACCCC(서열번호 45) | |
| DKRF | GCCAGTAGCGGCGGCGCTGGAAGGCGTAGGTGGGCAGATCTAGGTGGTGGTTGTGGGGGTG(서열번호 46) | Right arm (RA)의 제작 |
| DKRR | CTTAGGACCGTTATAGTTACGCGCTTCCTCGCTCACTGACTTTAATTAAAAGCTTACGTTTCGCTTGCCGATG(서열번호 47) |
5-3.
milA3
유전자에 양쪽 arm이
결합된
형태의 vector 제작
스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)로부터의 PCR을 통해 확보된 milA3영역을 포함하는 pM5-2벡터 및 pM6-4벡터를 제작하고, pM5-2벡터 에 제한효소 AfIII/BmtI를 처리하여 도14의 fragment 2를 얻었으며, pM6-4벡터에 제한효소 BstBI/SphI를 처리하여 도14의 fragment 4를 확보하였다.
상기 과정을 보다 구체적으로 기재하면 다음과 같다:
스트렙토마이세스 밀베마이시니쿠스 (S. milbemycinicus NRRL5739)의 milbemycin 생산유전자군을 클로닝하기 위하여, 스트렙토마이세스 빙쳉젠시스 (S. bingchenggensis BCW-1) 의 genome 염기서열 정보 (Accession no. CP002047.1)를 기반으로 milA2, milA3, milA4 지역을 포함하는 61kb의 영역을 각각 10kb 씩 6개 영역으로 나누어 프라이머를 디자인 하였다(표 7 및 도 19 참조)
| Fragment 명칭 | Primer Name | Sequence | 용도 |
| M1 | M1171-1NF | aaaatgcatCTACCGCCGCTTCAACAGC(서열번호 48) | Fragment 1 증폭 |
| M10810-1R | aaaactagtGGAGAGGAAGAAGGAGAGCCG(서열번호 49) | ||
| M2 | M10762-2NF | aaaatgcatCACCCACACAGGACAGGGA(서열번호 50) | Fragment 2 증폭 |
| MC20300-2R | aaaactagtATGACAGGCCGGGTACATCA(서열번호 51) | ||
| M3 | M20255-3NF | aaaatgcatCGCGTACCTGATGTCCCTGA(서열번호 52) | Fragment 3 증폭 |
| M30434-3R | aaaactagtGGCCACACGGTCTTCATCG(서열번호 53) | ||
| M4 | M30377-NF2 | aaaatgcatCCTGCACACCGACCGAGAC(서열번호 54) | Fragment 4 증폭 |
| M40160-4R | aaaactagtACCTTCGTACCCGTCTCGC(서열번호 55) | ||
| M5 | M40091-5NF | aaaatgcatGTGCGGACGATCGTGAGAAG(서열번호 56) | Fragment 5 증폭 |
| M50273-5R | aaaactagtAACACGCGGGAATCGACC(서열번호 57) | ||
| M6 | M50196-6NF | aaaatgcatGTAGTCCGTCGGCATCACAC(서열번호 58) | Fragment 6 증폭 |
| M60390-6R | aaaactagTCGAGACCGTCCCTATGTCC(서열번호 59) | ||
| M4N | M30290-F | ACGTCAGGAAGCGGTTCA(서열번호 60) | Fragment M4N 증폭 |
| M34887-R | CGTCCGAACGTTTCTGGA(서열번호 61) | ||
| M4CN | M4M4502F | GAGCTCCAGAAACGTTCGG(서열번호 62) | Fragment M4CN 증폭 |
| M4F7910R | GGATCGTCAACATCTCCTCC(서열번호 63) | ||
| M4CC | M4C-ScF | CGGCCACCAGAACACCTTC(서열번호 64) | Fragment M4CC 증폭 |
| M4C25R | ACACGCTTCTCACGATCGTC(서열번호 65) |
다음의 표 8의 조건으로 PCR 용 mixture를 제조하였다:
| Component | 1 reaction 당 (uL) | |
| 5X GXL buffer | 5 | Primer를 제외한 component를 모두 혼합 후 21 ul / tube 첨가 |
| 2.5 mM dNTPs | 2 | |
| S. milbemycinicus Genomic DNA (1/50 dil.) | 2 | |
| DW | 16.5 | |
| PrimeSTAR Pol. | 0.5 | |
| Primer for. (N), 5 pmol/ul | 2 | |
| Primer rev. (C), 5 pmol/ul | 2 |
상기 mixture에 대하여 MyCycler (Bio-Rad 사)를 사용하여 다음의 표 9의 조건에서 PCR을 수행하였다
| Step | Temperature | Time | Cycles |
| First denaturation | 95 ℃ | 5 min | 1 |
| Denaturation | 98 ℃ | 20 sec. | 30 |
| Extension | 68 ℃ | 10 min | |
| Final extension | 68 ℃ | 10 min | 1 |
증폭된 DNA fragment 중 milA3에 해당되는 fragment인 M5와 M6를 pCR2.1 TOPO cloning 시스템 (Invitrogen, 미국)을 이용하여 클로닝하고 E. coli DH5a를 형질전환하여 각각 pM5-2, pM6-4를 확보하였다.
상기 얻어진 fragments와, 상기 표 5의 프라이머 DK1F/DK1R, DK2F/DK2R, DK3F/DK3R 를 사용한 PCR 결과물 및 left, right flanking 영역과 단계적으로 overlap PCR 방법과 통상적인 ligation반응을 통하여 pLitmus28 (NEB, 미국) 에서 한 fragment로 연결하였다. 이와 같이 얻어진 재조합 벡터를 pYJ1957로 명명 하고, 이로부터 EcoRI/HindIII 제한효소 처리를 통해 얻은 left flanking region-milA3 모듈 7 DH~모듈 9 KR-right flanking region을 pKC1139로 옮겨 milA3 모듈 7 DH~모듈 9 KR 영역의 발현을 위한 재조합 벡터 (milA3 삽입 벡터; milA3 integration vector)를 제작하였다(도 14 참조; LA: left arm; RA: right arm; 1', 2, 3, 4, 5, 6: fragments). 이를 pD7K9milA3으로 명명하였다 (서열번호 80; LA: 6074-7261 부위; 1': 7256-10476 부위; 2: 10471-15043 부위; 3: 15038-17193 부위; 4: 17188-20776 부위; 5 및 6': 20771-21226 부위; RA: 21221-22436 부위).
실시예
6.
milA3
삽입 벡터(
milA3
integration vector)가 삽입된 균주 제작 및 생성물질 확인
상기 실시예 5-3에서 제작된 milA3 integration 벡터 (pD7K9milA3) 를 S. avertmilis DBM-01 균주 내로 도입시키고, 상동재조합이 일어나도록 유도하여 S. avertmilis DBM-01 균주의 aveA3 유전자가 milA3 유전자로 치환된 형태의 균주를 확보하고, 배양을 통해 생성되는 신규물질을 확인하였다.
6-1. 접합법을 통한 벡터 도입 및 유전자 치환
E. coli ET12567/pUZ8002 (The John Innes Foundation, 영국) 를 치환 벡터(pD7K9milA3)로 형질전환시킨 형질전환체를 아프라마이신 (apramycin), 클로람페니콜 (chloramphenicol), 카나마이신 (kanamycin)이 포함된 2xTY (trptone 16 g/L, yeast extract 10 g/L, NaCl 5 g/L) 또는 LB (peptone 10g/L, yeast extract 5g/L, sodium chloride 10g/L) 액체 배지를 이용하여 37 ℃에서 배양하였다. 12 시간 후, 1 ul의 배양액을 25 mL의 2xTY에 옮기고, 37℃에서 광학 밀도 (optical density)가 0.6이 될 때까지 배양하였다. 배양액의 세포를 동일한 액체 배지로 두 번 세척한 뒤, 500 uL의 2xTY로 현탁하여 얼음에 보관하였다. 스트렙토마이세스 아베미틸리스 DBM-01의 포자액 30-50 uL를 원심분리 (12,000 rpm으로 10분간)하고, 500 uL의 2xTY 에 현탁한 후, 50℃에서 10분 동안 열충격을 가하여 균사액을 준비하였다. 얼음에 보관해둔 E. coli 현탁액 500 uL과 상기 준비된 균사액 500 uL을 서로 혼합한 후 MS (agar 20g/L, D-mannitol 20g/L, soya flour 20g/L) 배지에 도포하여 28℃에서 24시간 동안 배양한 뒤 날리딕신산 (nalidixic acid)과 아프라마이신 (apramycin) 각 1mg을 배지위에 도말하였다. 7~10일 뒤 아프라마이신 저항성을 지닌 콜로니를 선별하고 37 ℃에서의 배양을 통해 항생제 저항성 여부를 재확인 후, 삼차 스트릭킹 (streaking)을 통해 얻은 콜로니에 대해 PCR 및 PCR 결과물에 대한 sequencing을 수행하여 일차교차 균주를 선별하였다.
상기 선별된 콜로니로부터의 계대배양을 통해 이차교차를 유도하고, 이를 아프라마이신이 첨가된 ISP4 고체배지와 아프라마이신이 첨가되지 않은 ISP4 고체배지에 각각 도말하여 아프라마이신 저항성을 잃은 콜로니를 선별하였다.
상기 사용된 ISP4 고체배지의 조성을 아래의 표 10에 나타내었다:
PCR 분석을 통해 최종 milA3가 치환된 이차교차 균주가 확보되었다. 상기 균주 제작 과정을 도 15에 모식적으로 나타내었다.
상기와 같이 확보된 균주를 S. avermitilis DBM-03-C로 명명하였다. S. avermitilis DBM-03-C 균주는 2016년 8월 24일자로 대한민국 전라북도 정읍시에 위치하는 한국생명공학연구원(Korean collection for type culture)에 기탁하여 수탁번호 KCTC13083BP를 부여받았다.
6-2.
milA3
치환균주의
배양
Milbemyci의 생산확인을 위한 종균배양 및 생산배양을 수행하였으며, 이 때 상용된 배지의 조성을 다음의 표 11에 나타내었다:
| Seed culture | g/L | Main culture | g/L |
| Soluble starch | 30.0 | Soluble starch | 80.0 |
| Yeast extract | 15.0 | Soybean flour | 10.0 |
| Corn steep liquor | 5.0 | Skim milk | 15.0 |
| KH2PO4 | 0.4 | KH2PO4 | 0.5 |
각 분량의 soluble starch, yeast extract, corn steep liquor, 및 KH2PO4를 DW에 녹여서 혼합한 후, NaOH를 사용하여 pH 7.2로 맞춘 후, 121℃에서 20 분간 고압살균하였다. ISP4 고체배지 (상기 표 5 참조)에 배양한 milA3 발현 균주 (S. avermitilis DBM-03-C)의 포자 혹은 mycelium을 긁어 종균배지에 접종한 뒤 28 ℃, 230rpm에서 40~48 시간 배양하여 종균배양액을 준비하였다. Soluble starch, soybean flour, skim milk, 및 KH2PO4를 DW에 녹이고 pH 7.2에 맞춰 제조한 생산배양 배지를 종균배양 배지와 같은 조건하에서 고압 멸균한 뒤 생산배양 배지 총 부피의 5 % 비율로 상기 준비된 종균배양액을 접종하여 230 rpm 및 28 ℃ 조건에서 10~15 일간 배양하였다.
6-3. 생성물질의 추출 및 분석
분석을 위한 시료의 전처리를 위하여, 상기 얻어진 배양액 0.5 mL을 동일 부피의 acetonitrile 및 methanol 혼합 용액 (acetonitrile:methanol=1:1 (v:v)) 또는 동일 부피의 ethyl acetate 와 혼합하여 2 분간 vortex 처리 한 뒤 10 분간 초음파 추출을 수행하였다. 5분간 원심분리하여 고형물을 제외하고 상층액만을 취하였다. 동일한 방법으로 추출을 3회 반복하여 분석시료를 확보하였다. 상기 확보된 시료는 HPLC 및 UPLC-qTOF-HR-MS를 통해 분석하였다.
HPLC는 PDA detector와 gradient pump로 구성된 HPLC system (YL Instrument Co. Ltd)하에서 Phenomenex Luna (4.6x100 mm, 5 mm) column을 이용하여 수행하였고, 물 기반의 0.05% (v/v) trifluoroacetic acid (TFA)를 용매 A로 하고 acetonitrile을 용매 B 로 하고 표 12와 같은 용매 기울기를 사용하여, 238~245 nm 에서 1 mL/min 의 flow rate로 총 60 분간 시료를 분석하였다.
Milbemycin A3/A4의 30%/70% (w/w) 표준품인 milbemectin (Bioaustralis, Australia) 및 milbemycin D (Bioaustralis, Australia)를 사용하여 본 HPLC 분석 조건에서의 해당 milbemycin의 retention time (RT) 및 245 nm에서의 absorbance를 확인하였다. 상기 확보된 시료 (S. avermitilis DBM-03-C 균주)의 생성 물질을 분석한 결과를 도 16에 나타내었다. 도 16에서, (a)는 표준폼 milbemectin (Bioaustralis, Australia) 및 milbemycin D (Bioaustralis, Australia)의 분석 결과를, (b)는 S. S. avermitilis DBM-03-C 균주의 생성물질 분석 결과를 각각 보여준다. 도 16에 나타난 바와 같이, 상기 S. avermitilis DBM-03-C 균주는 milbemycin A3 (CAS NO. 51596-10-2), A4 (CAS NO. 51596-11-3), alpha2 (CAS No. 51596-12-4), D (CAS NO. 77855-81-3), alpha4 (CAS No. 51596-13-5), 및 G (CAS No. 83471-31-2)로 추정되는 피크를 확인할 수 있고, 이 중에서, milbemycin A3, A4, 및 D는 표준품 milbemycin과 동일한 RT값을 지님을 확인할 수 있다. 특히 milbemycin A3, A4 및 D로 추정되는 peak는 표준품 milbemycin 과 동일한 RT 값을 지님을 알 수 있다. 도 16의 HPLC 분석 결과에 나타난 각 물질의 RT는 다음과 같다:
Milbemycin A3 RT: 29.0 min
Milbemycin A4 RT: 32.1 min
Milbemycin D RT: 35.4 min
Milbemycin alpha 2 RT: 33.7 min
Milbemycin alpha 4 RT: 36.3 min
Milbemycin G RT: 39.4 min.
HPLC 상에서 milbemycin으로 추정되는 물질의 분자량을 비교하기 위하여, UPLC-qTOF-HR-MS를 다음과 같이 수행하였다. 상기 HPLC 상에서 분석된 여섯 종류의 milbemycin의 해당 분자량 값을 Xselect® CSH column XP (2.1x100 mm, 2.5 um; Waters) 을 통해 Waters XEVO® G2S Q-TOF mass spectrometer; Waters) 상에서 확인하였다. 이 때 사용된 용매 기울기는 아래의 표 13과 같이 하였다.
상기 얻어진 질량 스펙트럼 결과를 도 17a 내지 17f에 나타내었다 (도 17a: 밀베마이신 A3; 도 17b: 밀베마이신 A4; 도 17c: 밀베마이신 D; 도 17d: 밀베마이신 alpha2; 도 17e: 밀베마이신 alpha4; 도 17f: 밀베마이신 G). 도 17a 내지 17f에 나타난 바와 같이, milbemycin A3에 해당하는 m/z=527.3014, milbemycin A4 및 α2에 상응하는 m/z=541.3171, milbemycin D및 α4를 나타내는 m/z=555.3327, milbemycin G의 값인 m/z=569.3484을 negative ion mode에서 확인할 수 있었으며, 각각 분자의 알려진 분자량과 잘 일치하는 것으로 보아 제작된 재조합 스트렙토마이세스 아베르미틸리스 DBM-03-C 가 밀베마이신을 생산함을 알 수 있었다.
<110> Dongbu Farm Hannong Co., Ltd.
Ewha University - Industry Collaboration Foundation
<120> Recombinant microorganism producing milbemycin and method for
producing milbemycin using the same
<130> DPP20163328KR
<150> KR10-2015-0133892
<151> 2015-09-22
<160> 80
<170> KopatentIn 1.71
<210> 1
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> A1NT2-F primer
<400> 1
gaaccgtgtg cggaagaa 18
<210> 2
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> A1CT1-R primer
<400> 2
atcaggtcgg tgagacggt 19
<210> 3
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> A1CT1-F primer
<400> 3
gcaaggtcgt ctttgtgtgc 20
<210> 4
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> A1CT2-R primer
<400> 4
ccaccaactc gatatgggc 19
<210> 5
<211> 17
<212> DNA
<213> Artificial Sequence
<220>
<223> A2-404F primer
<400> 5
ggcgctggca atggacc 17
<210> 6
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> A2PH-1500R primer
<400> 6
gattacgcca agcttggctg attctcccgg gacg 34
<210> 7
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> AveD2-F primer
<400> 7
gatgacggaa atcccctgg 19
<210> 8
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> AveD2-R primer
<400> 8
agaatgaaag gagcgcga 18
<210> 9
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> MA1NU-F primer
<400> 9
gaggacacag ctcagttgcc caaagcccag aac 33
<210> 10
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> M1Age-R primer
<400> 10
tgatcgaaga gccgggagt 19
<210> 11
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> PHR-M1Ag primer
<400> 11
ccggctcttc gatcaagctt ggcgtaatca tgg 33
<210> 12
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> PXF-AD2 primer
<400> 12
gggatttccg tcatcaagct tggcgtaatc atgg 34
<210> 13
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> aveFi-F primer
<400> 13
ggcaacacca accgtgagat 20
<210> 14
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> milA1i-R primer
<400> 14
gatgaccgag gagtggaacg 20
<210> 15
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> milA1i-F primer
<400> 15
gtactggacc tgttcgagcg 20
<210> 16
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> aveA2i-R2 primer
<400> 16
atgaatgcgg tctttcctgc 20
<210> 17
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> aveA1C-F primer
<400> 17
ggactcgctg attcggacaa 20
<210> 18
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> aveA1C-R primer
<400> 18
ggagcagatg aatgccctct 20
<210> 19
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> milA1C-F primer
<400> 19
cctgtgggag ttcatcaccg 20
<210> 20
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> milA1C-R primer
<400> 20
cgagattgtg gtggacgagg 20
<210> 21
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> MA3-6503R primer
<400> 21
tgccaatgat cgcgatggcc tcatc 25
<210> 22
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> MA3-104Fin(A) primer
<400> 22
gggtcagtgg cacgaacgcc catgtgatcc tcg 33
<210> 23
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> MA3-1368Ffu(B) primer
<400> 23
ggaggcgtcc agcgaaccca tcgccatcat c 31
<210> 24
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> AA3-8600RH primer
<400> 24
aaaaagctta acggtgtgtg gagcgtcag 29
<210> 25
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> AA3-5597FSp primer
<400> 25
aaaactagtc gcgatcattg gcatg 25
<210> 26
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> AA3-111RSp primer
<400> 26
aaaactagtg ctggacgcct ccatggc 27
<210> 27
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> AA3-1369RSp primer
<400> 27
aaaactagtc gtgccactga ccccgaac 28
<210> 28
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> AA4-2300FNs primer
<400> 28
aaatctagag ctccgtgcag accgagaac 29
<210> 29
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> aac3iv-F primer
<400> 29
ggttcatgtg cagctccatc 20
<210> 30
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> aac3iv-R primer
<400> 30
cgtcgcggtg agttcagg 18
<210> 31
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Amo7i-F primer
<400> 31
gttgtcggtg ttgcttccg 19
<210> 32
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Amo7i-R primer
<400> 32
cacatcacac gcaacaatcc 20
<210> 33
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Mmo7i-F primer
<400> 33
acatcagcgg cctctacga 19
<210> 34
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Mmo7i-R primer
<400> 34
ccaggtccat cactccctc 19
<210> 35
<211> 1005
<212> DNA
<213> Artificial Sequence
<220>
<223> pGEMB2-MA-DOWN-fu (Fig. 3)
<400> 35
ggatccagga cccggcggac accccccgcg gccgccgcca ccgggccgtt ccccctgcat 60
acccgcctcg gcggtcacgc cccggccgag cagcacgaga tgctgctctc gctggtccgc 120
tcccacgccg ccctcgtgct gggccgcgac gatccggaca cggtccatcc cggcgcgcac 180
ttccgcggcc tgggcttcga ctccctgacc gcggtcgagc tccgcaaccg gctcaacgcc 240
gccaccggcc tccggctctc caccaccctc gtcttcgacc accccacgcc cgacgaactc 300
gcccgtcacg tccgggagca ggtgctgggc gacggcgaag cggcgcgggt ggcccccgtg 360
ttggcggagc tcgacaaact cgaatccgcc ctctccgccc tcgacaaaac cgacagcgcc 420
agcgaaagag tcaccctgcg gctgaagtca ctcatgttga ggtggaacgc accccagcat 480
ccgacagccg aaagcgctga tgacgacgag aagttcacat cggcaacaga ggctgagatt 540
ttcaaattca ttgacaacga cctcggcctg tcctgaaccg gacgcctgcc actccgcccg 600
tatccgctgg gccctgctag gacgtgaatg caattggcga atgaagcgaa gctcctggaa 660
tacctcaagc gcgtcactgc ggacctggac cgcactcgcc gtcgcctgta cgaggtggtc 720
gagcgtgagc aggagccgat cgcgattgtg gggatggcgt gtcgttaccc aggcggggcg 780
acgtcaccca cgcgactgtg gcatctcgtc aagtcccaga cggacgctat cggggagttc 840
ccgaccgacc gtggatggaa cctggagcag ctctacgacc cggaccccga ccgctcagga 900
accagttaca cgcgcagcgg agggtttctc tatgacgcgg gcgacttcga cgccgcgttc 960
ttcgagttgt caccgcgtga ggcgctggca atggaattca agctt 1005
<210> 36
<211> 191
<212> PRT
<213> Artificial Sequence
<220>
<223> MilA1 sequence (module 2) and AveA1 sequence (Docking domain) in
pGEMB2-MA-DOWN-fu (Fig. 3)
<400> 36
Gly Ser Arg Thr Arg Arg Thr Pro Pro Ala Ala Ala Ala Thr Gly Pro
1 5 10 15
Phe Pro Leu His Thr Arg Leu Gly Gly His Ala Pro Ala Glu Gln His
20 25 30
Glu Met Leu Leu Ser Leu Val Arg Ser His Ala Ala Leu Val Leu Gly
35 40 45
Arg Asp Asp Pro Asp Thr Val His Pro Gly Ala His Phe Arg Gly Leu
50 55 60
Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Asn Ala
65 70 75 80
Ala Thr Gly Leu Arg Leu Ser Thr Thr Leu Val Phe Asp His Pro Thr
85 90 95
Pro Asp Glu Leu Ala Arg His Val Arg Glu Gln Val Leu Gly Asp Gly
100 105 110
Glu Ala Ala Arg Val Ala Pro Val Leu Ala Glu Leu Asp Lys Leu Glu
115 120 125
Ser Ala Leu Ser Ala Leu Asp Lys Thr Asp Ser Ala Ser Glu Arg Val
130 135 140
Thr Leu Arg Leu Lys Ser Leu Met Leu Arg Trp Asn Ala Pro Gln His
145 150 155 160
Pro Thr Ala Glu Ser Ala Asp Asp Asp Glu Lys Phe Thr Ser Ala Thr
165 170 175
Glu Ala Glu Ile Phe Lys Phe Ile Asp Asn Asp Leu Gly Leu Ser
180 185 190
<210> 37
<211> 122
<212> PRT
<213> Artificial Sequence
<220>
<223> AveA2 sequence in pGEMB2-MA-DOWN-fu (Fig.3)
<400> 37
Met Gln Leu Ala Asn Glu Ala Lys Leu Leu Glu Tyr Leu Lys Arg Val
1 5 10 15
Thr Ala Asp Leu Asp Arg Thr Arg Arg Arg Leu Tyr Glu Val Val Glu
20 25 30
Arg Glu Gln Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg Tyr Pro
35 40 45
Gly Gly Ala Thr Ser Pro Thr Arg Leu Trp His Leu Val Lys Ser Gln
50 55 60
Thr Asp Ala Ile Gly Glu Phe Pro Thr Asp Arg Gly Trp Asn Leu Glu
65 70 75 80
Gln Leu Tyr Asp Pro Asp Pro Asp Arg Ser Gly Thr Ser Tyr Thr Arg
85 90 95
Ser Gly Gly Phe Leu Tyr Asp Ala Gly Asp Phe Asp Ala Ala Phe Phe
100 105 110
Glu Leu Ser Pro Arg Glu Ala Leu Ala Met
115 120
<210> 38
<211> 50
<212> DNA
<213> Artificial Sequence
<220>
<223> DK1F primer
<400> 38
gcgtccgggg tcgacatggg atgcaggcct gagcggttga tgcgggcccg 50
<210> 39
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> DK1R primer
<400> 39
atgggactgc ggctgcttaa gacctcaccc gtc 33
<210> 40
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> DK2F primer
<400> 40
gctagcgggg tgtggtcg 18
<210> 41
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> DK2R primer
<400> 41
ttcgaacacg cgggaatcga c 21
<210> 42
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> DK3F primer
<400> 42
gcatgccgtg tccacgcgcc c 21
<210> 43
<211> 49
<212> DNA
<213> Artificial Sequence
<220>
<223> DK3R primer
<400> 43
acacccaccc ccacaaccac cacctagatc tgcccaccta cgccttcca 49
<210> 44
<211> 53
<212> DNA
<213> Artificial Sequence
<220>
<223> DKLF primer
<400> 44
aacccgaacg accgagcgca gcggcggccg cgaattcata cggctacgag gat 53
<210> 45
<211> 59
<212> DNA
<213> Artificial Sequence
<220>
<223> DKLR primer
<400> 45
ctcgaccgca ccgaccgggc ccgcatcaac cgctcaggcc tgcatcccat gtcgacccc 59
<210> 46
<211> 61
<212> DNA
<213> Artificial Sequence
<220>
<223> DKRF primer
<400> 46
gccagtagcg gcggcgctgg aaggcgtagg tgggcagatc taggtggtgg ttgtgggggt 60
g 61
<210> 47
<211> 73
<212> DNA
<213> Artificial Sequence
<220>
<223> DKRR primer
<400> 47
cttaggaccg ttatagttac gcgcttcctc gctcactgac tttaattaaa agcttacgtt 60
tcgcttgccg atg 73
<210> 48
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> M1171-1NF primer
<400> 48
aaaatgcatc taccgccgct tcaacagc 28
<210> 49
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> M10810-1R primer
<400> 49
aaaactagtg gagaggaaga aggagagccg 30
<210> 50
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> M10762-2NF primer
<400> 50
aaaatgcatc acccacacag gacaggga 28
<210> 51
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> MC20300-2R primer
<400> 51
aaaactagta tgacaggccg ggtacatca 29
<210> 52
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> M20255-3NF primer
<400> 52
aaaatgcatc gcgtacctga tgtccctga 29
<210> 53
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> M30434-3R primer
<400> 53
aaaactagtg gccacacggt cttcatcg 28
<210> 54
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> M30377-NF2 primer
<400> 54
aaaatgcatc ctgcacaccg accgagac 28
<210> 55
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> M40160-4R primer
<400> 55
aaaactagta ccttcgtacc cgtctcgc 28
<210> 56
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> M40091-5NF primer
<400> 56
aaaatgcatg tgcggacgat cgtgagaag 29
<210> 57
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> M50273-5R primer
<400> 57
aaaactagta acacgcggga atcgacc 27
<210> 58
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> M50196-6NF primer
<400> 58
aaaatgcatg tagtccgtcg gcatcacac 29
<210> 59
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> M60390-6R primer
<400> 59
aaaactagtc gagaccgtcc ctatgtcc 28
<210> 60
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> M30290-F primer
<400> 60
acgtcaggaa gcggttca 18
<210> 61
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> M34887-R primer
<400> 61
cgtccgaacg tttctgga 18
<210> 62
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> M4M4502F primer
<400> 62
gagctccaga aacgttcgg 19
<210> 63
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> M4F7910R primer
<400> 63
ggatcgtcaa catctcctcc 20
<210> 64
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> M4C-ScF primer
<400> 64
cggccaccag aacaccttc 19
<210> 65
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> M4C25R primer
<400> 65
acacgcttct cacgatcgtc 20
<210> 66
<211> 11919
<212> DNA
<213> Artificial Sequence
<220>
<223> aveA1 gene of Streptomyces avermitilis MA-4680
<400> 66
gtgcagagga tggacggcgg ggaagaaccc cgccctgcgg caggggaggt cctcggagtg 60
gccgacgagg cggacggcgg cgtcgtcttc gtttttcccg ggcagggccc gcaatggccg 120
ggcatgggaa gggaacttct cgacgcttcc gacgtcttcc gggagagcgt ccgcgcctgc 180
gaagccgcgt tcgcgcccta cgtcgactgg tcggtggagc aggtgttgcg ggactcgccg 240
gacgctcccg ggctggaccg ggtggacgtc gtccagccga ccctgttcgc cgtcatgatc 300
tccctggccg ccctctggcg ctcgcaaggg gtcgagccgt gcgcggtgct gggacacagc 360
ctgggcgaga tcgcggcagc ccacgtctcg ggaggcctgt ccctggccga cgccgcacgc 420
gtggtgacgc tttggagcca ggcacagacc acccttgccg ggaccggcgc gctcgtctcc 480
gtcgccgcca cgccggatga gctcctgccc cgaatcgctc cgtggaccga ggacaacccg 540
gcgcggctcg ccgtcgcagc cgtcaacgga ccccggagca cagtcgtttc cggtgcccgc 600
gaggccgtcg cggacctggt ggccgacctc accgccgcgc aggtgcgcac gcgcatgatc 660
ccggtggacg ttcccgccca ctcccccctg atgtacgcca tcgaggaacg ggtcgtcagc 720
ggcctgctgc ccatcacccc acgcccctcc cgcatcccct tccactcctc ggtgaccggc 780
ggccgcctcg acacccgcga gctagacgcg gcgtactggt accgcaacat gtcgagcacg 840
gtccggttcg agcccgccgc ccggctgctt ctgcagcagg ggcccaagac gttcgtcgag 900
atgagcccgc acccggtgct gaccatgggc ctccaggagc tcgccccgga cctgggcgac 960
accaccggca ccgccgacac cgtgatcatg ggcacgctgc gccgcggcca gggcaccctg 1020
gaccacttcc tgacgtctct cgcccaacta cgggggcatg gtgagacgtc ggcgaccacc 1080
gtcctctcgg cacgcctgac cgcgctgtcc cccacgcagc agcagtcgct gctcctggac 1140
ctggtgcgcg cccacaccat ggcggtgctg aacgacgacg gaaacgagcg caccgcgtcg 1200
gatgccggcc catcggcgag tttcgcccac ctcggcttcg actccgtcat gggtgtcgaa 1260
ctgcgcaacc gcctcagcaa ggccacgggc ctgcggttgc ccgtgacgct catcttcgac 1320
cacaccacgc cggccgcggt cgccgcgcgc cttcggaccg cggcgctcgg ccacctcgac 1380
gaggacaccg cgcccgtacc ggactcaccc agcggccacg gaggcacggc agcggcggac 1440
gacccgatcg ccatcatcgg catggcatgc cgtttcccgg gcggagtccg gtccccgaag 1500
gacctgtggg agctggccgc ctcgggcgga gacgccatcg ggccgttccc caccgaccgc 1560
ggatggccca cggaacagcg tcacgcccag gaccccacgc agcccggcac gttctatccg 1620
cagggaggcg ggttccttca cgacgcggcg cacttcgacg ccggcttctt cggaatcagt 1680
ccacgtgagg cactggcgat ggatccgcag cagcggctgc tgctggagac gtcctgggag 1740
gcgttcgagc gggcgggaat cgatccgctg tcggtacgcg ggtcccgtac gggcgtcttc 1800
gcgggcgccc tctccttcga ctacggcccg cgtatggaca ccgcgtcgtc ggagggcgcc 1860
gcggacgtgg agggccacat cctcaccggt accacgggca gcgtcctgtc gggccgtatc 1920
gcctacagct tcgggctgga agggccggcg atcaccgtgg acacggggtg ctcggcatcg 1980
ctcgtgacgc tgcatctggc gtgccagtcg ctgcggtcgg gtgagtgcac gctcgcgctg 2040
gccggcggcg tctcggtcat gtccaccctc ggcatgttca tcgagttctc ccggcagcgc 2100
gggctgtcgg tggacggcag gtgcaaggcg tactcggctg cagccgacgg caccggctgg 2160
ggcgagggcg tcgggatgct gttggtggag cggttgtcgg atgcggtgcg gctggggcat 2220
cgggtgctgg cggtggtacg cggcagtgcg gtcaaccagg acggtgcgtc gaatgggctg 2280
acggcgccga acggtccggc tcaggagcgg gtgatccggc aggcgttggc gaacgcgggg 2340
ttgtccgtgg cggatgtgga tgtggtggag gggcacggga cgggcacgac gctgggtgat 2400
ccgatcgagg cacaggcgtt gctcgccacg tacgggcagc gggccggtga caggccgctg 2460
tggctggggt ctctgaagtc caacatcggg cacaccatgg ctgccgcggg tgtgggtggg 2520
gtcatcaaga tggtgatggc gttgcgggag ggggtgttgc cgcggacgtt gcatgtggat 2580
aagccgtcgc cgcaggtgga ctggtccgcg ggggcggtgc ggctgctgac ggaggcggtg 2640
ccgtggccgg gggacgcggc agggcggttg cggcgggcgg gagtgtcgtc gttcgggatc 2700
ggcggcacga atgcgcatgt gattttggag gaggcgccgg cggcgggggg ctgtgttgcc 2760
gggggtgggg tgttggaggg tgctccgggt cttgccattt cggtggctga gtcggtggcc 2820
gctccagtgg ctgtgtctgc gccggtggct gagtcggtgc cggtgccggt gccggtgccg 2880
gttcctgtgc cggtgtcggc taggtctgag gctgggttgc gggcgcaggc ggaggcgttg 2940
cgtcagtacg tggcagtccg gccggacgtt tcgcttgccg atgtgggtgc gggtctggcc 3000
tgtgggcggg ctgtgctgga gcatcgtgcg gtcgtcctgg ccgcggaccg tgaggagctg 3060
gtgcaagggt tgggggcgct ggcggcgggt gagccggatc ggcgggtgac cacgggtcat 3120
gcgccgggtg gtgaccgggg cggtgtcgtc ttcgtgtttc ccggacaggg tgggcagtgg 3180
gccgggatgg gtgtgcgtct gctcgcctcc tctccggtgt tcgcccggcg gatgcaggcg 3240
tgcgaggagg ctctggcgcc gtgggtggac tggtctgtgg tggacatcct gcgccgggac 3300
gcgggggatg cggtgtggga gcgggccgat gtggtccagc ctgtgctgtt cagcgtcatg 3360
gtgtctttgg ctgctctgtg gcgttcctac ggtatcgaac ccgacgcggt ccttggccat 3420
tcccagggcg agatcgcggc cgcgcatgtg tgtggggcgc tgagcctgaa ggacgcggcg 3480
aagactgttg cgctgcgcag ccgggcgctg gccgctgtgc ggggccgggg cggcatggcc 3540
tcagtgccgc tgcctgccca ggaggtggag cagctcattg gtgagcggtg ggcggggcgg 3600
ttgtgggtgg cggcggtcaa cggcccccgc tccaccgccg tctcggggga tgccgaggcg 3660
gtggacgagg tgctggcgta ctgtgccggc accggggtgc gggcccggcg gatcccggtc 3720
gactatgcct cgcactgccc ccatgtgcag cccctgcggg aggagttgct ggagctgctg 3780
ggggacatca gcccgcagcc gtccggcgtg ccgttcttct ccacggtgga gggcacctgg 3840
ctggacacca caaccctgga cgccgcctac tggtaccgca acctgcacca gccggtccgt 3900
ttcagcgatg ccgtccaggc cctggcggat gacggacacc gcgtcttcgt cgaagtcagc 3960
ccccacccca ccctcgtccc cgccatcgaa gacaccaccg aagacaccgc cgaagacgtc 4020
accgcgatcg gcagcctccg ccgcggcgac aacgacaccc gccgcttcct caccgccctc 4080
gcccacaccc ataccaccgg catcggcaca cccaccacct ggcaccacca ctacacccac 4140
caccacaccc acccccaccc ccacacgcac ctcgacctgc ccacctaccc cttccaacac 4200
cagcactact ggctcgagag ctcacagccg ggtgccggat ccggttcggg tgccggtgcc 4260
ggttcgggtg ccggttccgg gcgggcaggg actgcgggcg ggacggcaga ggtggagtcg 4320
cggttctggg acgcggtggc ccgccaggac ctggaaacgg tcgcgaccac actcgccgtg 4380
cccccctccg ccggcctgga cacggtggtg cccgcactct ccgcctggca ccgccaccaa 4440
cacgaccaag cccgcatcaa cacctggacc taccaggaaa cctggaaacc cctcaccctc 4500
cccaccaccc accaacccca ccaaacctgg ctcatcgcca tccccgaaac ccagacccac 4560
cacccccaca tcaccaacat cctcaccaac ctccaccacc acggcatcac ccccatcccc 4620
ctcaccctca accacaccca caccaacccc caacacctcc accacaccct ccaccacacc 4680
cgacaacaag cccaaaacca caccaccgga gccatcaccg gcctgctctc cctcctcgcc 4740
ctcgacgaaa caccccaccc ccaccacccc cacacaccca ccggcaccct cctcaacctc 4800
accctcaccc aaacccacac ccaaacccac ccaccaaccc ccctctggta cgccaccacc 4860
aacgccacca ccacccaccc caacgacccc ctcacacacc ccacccaagc ccaaacctgg 4920
ggactcgccc gcaccaccct cctcgaacac cccacccaca ccgccggaat catcgacctc 4980
cccaccaccc ccacccccca caccctccag cacctcaccc aaaccctcac ccaaccccac 5040
caccaaaccc aactcgccat ccgcaccacc ggcacccaca cccgccgcct cacccccacc 5100
accctcaccc ccacacacca accacccacc cccacccccc acggaaccac cctcatcacc 5160
ggcggaaccg gcgccctcgc cacccacctc acccaccacc tcaccaccca ccaacccacc 5220
caacacctcc tcctcaccag ccgaaccggc ccccacaccc cccacgcaca acacctcacc 5280
acccaactcc aacaaaaagg catccacctc accatcacca cctgcgacac cagcaaccca 5340
gaccaactcc aacaactcct caacaccatc cccccacaac accccctcac caccgtcatc 5400
cacaccgcag gcatcctcga cgacgccacc ctcaccaacc tcacccccac ccaactcaac 5460
aacgtcctcc gcgccaaagc ccacagcgcc cacctcctcc accaactcac ccaacacacc 5520
cccctcaccg ccttcgtcct ctactcctcc gccgccgcca ccttcggcgc acccggccaa 5580
gccaactacg ccgcagccaa cgcctacctc gacgccctcg cccaccaccg ccacacccac 5640
cacctccccg ccaccagcat cgcctggggc acctggcaag gaaacggact cgctgattcg 5700
gacaaggccc gcgcatatct cgaccgccgc gggtttcgac ccatgtcacc cgagttggcc 5760
acggcagcgg tcacgcaggc gatcgcggac accgaacggc cgtatgtcgt catcgccgac 5820
atcgactgga gcaagatcga acacacctct cagaccagcg acctggtgag cgcggcccgg 5880
gaaagggagc cagctgtcca gcgccccact ccaccggcgg agttgcacaa aacgctggcc 5940
catcagacgt cggccgacca acgggccgca ttgctcgagc tcgtacgaga ccatgtggcg 6000
gcagtgctcc ggcacgcgga cccgaaagcc atcgcgcccg accagtcgtt ccgtgcactc 6060
ggcttcgatt cactcacggc cgtcgagttc cgaaacctgc tgatcaaggc aacaggactc 6120
cgccttcctg tctcgctggt cttcgaccac ccgacccctg ccaaactcgc cgtacacctg 6180
cagaaccaac tgcggggcac agcagcggag tcggctcctt cagcggcagc cgttaccgcc 6240
gaggcttctg tcaccgagcc gatcgccatc gttggcatgg cctgtcgttt ccccggcgga 6300
gtgacctcgg cggacgactt ctgggatctg atctcctccg agcaggacgc gatcggcgga 6360
ttccccaccg accgcggctg ggacctggac acgctctacg accccgaccc cgaccacccc 6420
ggcacctgct acacccgaaa cggcggattc ctctacgacg caggccactt cgacgccgaa 6480
ttcttcggca tcagcccccg cgaagccctc gccatggacc cccagcaacg actcctcctc 6540
gaaaccgcct gggaaaccat cgaacacgcc ggcatcaacc cccacaccct ccacggcacc 6600
cccaccggag tcttcaccgg caccaacgga caggactacg cacttcgcgt gcacaacgcg 6660
ggccagtcaa ccgatggttt cgcactgacc ggaaccgccg gcagcgtcat ctccggtcgt 6720
atctcgtaca cgtttggttt tgagggtcct gcggtgtcgg tggacacggc ttgttcctcg 6780
tcgttggtgg ctttgcatct ggcctgtcag gcgttgcgtg cgggtgagtg ctcgatggcg 6840
cttgccgggg gtgtgacggt gatgtcgtct ccgggtgcct tcgtggagtt ttcgcggcag 6900
cggggtctgg ccgcggacgg gcattgcaag gcgttctcgg cggcggcgga cgggaccggc 6960
tggggtgagg gtgtggggat gctgctggtg gagcggctct ccgacgccca tcgcaacggt 7020
caccgtgtcc tggccgtggt gcgtggcagt gcggtcaacc aggacggtgc gagcaacggt 7080
ctgaccgcgc ccaacgggcc gtcccagcag cgtgtcatcc gccaggccct cgccaacgcc 7140
ggcttgtcgg ccggtgatgt cgacgcggtg gaggcccacg gcaccggcac cactttgggc 7200
gacccgatcg aggcccaggc cctcctcgcg acctacggac aggaccgtgc cggcgagggg 7260
ccgctgtggc tgggctcggt caagtccaat gtcggtcaca cacaggctgc cgcgggcgtc 7320
gccggggtga tcaagatggt gatggcgctg cggcatggtc tgctgccgcg gacgttgcat 7380
gtggatgagc cgtcgccgca tgtggactgg tccgcgggtg cggtgcagct gctgacggag 7440
acggtgccct ggcccggcgg ggaggggcgg ctacggcggg caggagtgtc atcattcggc 7500
gtcagcggca ccaacgccca cgtcatcctc gaagaagcac ccgccgacga cgttccgggg 7560
ggaccacccg ccggcgaggg tgacgcgggc agcgacgatg aggctgctgc cggcagtcct 7620
ggggtgtggc cgtggctggt gtcggccaag tcgcagccgg ccctgcgcgc ccaggcccag 7680
gccctgcacg cccacctcac cgaccacccc ggcctcgacc tcgcggatgt cggatacacc 7740
ctcgcccacg cccgcgccgt gttcgaccac cgcgccaccc tcatcgccgc ggaccgcgac 7800
acgttcctgc aagcactcca ggcactcgcc gcaggcgagc cccaccccgc cgtcatccac 7860
agcagcgccc cgggcgggac cgggaccggg gaggccgcag gaaagaccgc attcatctgc 7920
tccggacagg gcacccaacg ccccggcatg gcccacggcc tctaccacac ccaccccgtc 7980
ttcgccgccg cactcaacga catctgcacc cacctcgacc cccacctcga ccaccccctc 8040
ctccccctcc tcacccaaaa cgacaacgac aacgaggacg cggccgcact gctccagcag 8100
acccgctacg cccagcccgc cctcttcgcc ttccaggtcg ccctccaccg cctcctcacc 8160
gacggctacc acatcacccc ccactactac gccggacact ccctcggcga aatcaccgcc 8220
gcccacctcg ccggcatcct caccctcacc gacgccacca ccctcatcac ccaacgcgcc 8280
accctcatgc aaaccatgcc ccccggcacc atgaccaccc tccacaccac cccccaccac 8340
atcacccacc acctcaccgc ccacgaaaac gacctcgcca tcgccgccat caacaccccc 8400
acctccctcg tcatcagcgg caccccccac accgtccaac acatcaccac cctctgccaa 8460
caacaaggca tcaaaaccaa aaccctcccc accaaccacg ccttccactc cccccacacc 8520
aaccccatcc tcaaccaact ccaccagcac acccaaaccc tcacctacca cccaccccac 8580
acccccctca tcaccgccaa caccccaccc gaccaactcc tcacccccca ctactggacc 8640
caacaagccc gcaacaccgt cgactacgcc accaccaccc aaaccctcca ccaacacggc 8700
gtcaccacct acatcgaact cggacccgac aacaccctca ccaccctcac ccaccacaac 8760
ctccccaacc cccccaccac caccctcacc ctcacccacc cccaccacca cccccaaacc 8820
cacctcctca ccaacctcgc caaaaccacc accacctggc acccccacca ctacacccac 8880
cacgacaacc aaccccacac ccacacccac ctcgacctcc ccacctaccc cttccaacac 8940
caccactact ggctcgaaag cacacagccc ggtgccggca acgtgtcagc agccggactc 9000
gaccccaccg aacaccccct actcggcgcc acattggaac tggcgactga cggtggagcg 9060
cttcttgcag ggcgcttgtc tttgaggtcg catccgtggc tggctgacca tgccgtcggc 9120
ggcacggtgc tgctgtcggg cgccaccttc ctcgaactcg cccttcatgc gggcacatac 9180
gtgggctgcg accgagtgga tgagctgacg ctgcatgcgc cgctggtggt tcctgtggat 9240
gggggtgtga gtgtgcaggt tggggttgcg gctgcggatg gggaggggcg gcgtttggtg 9300
agtgtgtatg cgcggggtgg gagtgcttgt ggtgggggtg gtgcgtcggg tggggtgtgg 9360
acgtgtcatg cctcgggggt gctggttgag gctgctgctg gtggtgtggt ggtggatggt 9420
ctggcggggg tgtggccgcc gcggggtgcg gtggcggtgg atgtcgatgg tgtccgtgac 9480
cgtttggctg gggctggttg tgttttgggg ccggtgtttt cggggctgcg tgcggtgtgg 9540
cgtgatgggg gggatttgct ggctgaggtg tgtctgccgg aggaggcgtg gggtgatgcg 9600
gctggttttg ggctgcatcc ggcgttgctg gatggtgtgg tccagccgtt gtcggtgttg 9660
cttccgggtg ggacggggtt tggggagggg gcggggttcg gggagggtgt tcgggtgccg 9720
gctgtgtggg gtggtgtgtc gcttcaccgg gcgggtgtga ccggtgtgcg ggtgcgtgtg 9780
tcggctgtcg ggcggggcgg cgggcgtgag gcggtgtcgg tcgtggtcgg ggatgaggcg 9840
ggtgtgccgg tggcgtcggt cgatcgtctt gagttgcggc ctgtggatat gggtcagttg 9900
cgtgctgtct cggtttcggc ggggcggcgg ggttcgctgt atgcggtgca gtgggctgag 9960
gtgggtcctg tgccggtgtg tgggcaggcg tgggcgtggc acgaggacgt gggtgagagc 10020
ggtggtgggc ctgtgccggg ggtggtggtg ttgcggtgcc cggatgccgg tgccggtggc 10080
ggtggcggtg gcggtggtgg cggtggtgtg ggtgaggttg ttggtggggt gttgggtgtg 10140
gtgcaggggt ggctggggct ggagcggttt gcgggttcgc ggctggtggt ggtgacccgg 10200
ggtgcggtgg tggccggccc ggaggacggc ccggtggatg tggtgggtgc gtcggtgtgg 10260
gggctggtgc gttcggcgca ggctgagcat ccggaccggt ttgtcctcct cgacctcgac 10320
accgacaccg gcaccgacct cgacaccggt gctggtgctg gttggggcgt ggatggtggg 10380
cgtgtggcgg cggtggtggc gtgtggtgag ccgcagttgg cggtgcgtgg ggagcggttg 10440
ctggccgcac gcctgaaacg acttgagtca tccggtgatg ttccagccca gcggtccggt 10500
gacacacgag cccggcggtc cgacgtgcct gcccagcgct ccggtggcgt gcctgctcgg 10560
cggtcggttg atgtatcggg tcgggaggtg ttgccgtggt tgtcgggtgg gtcggtgttg 10620
gtgacgggtg ggacgggtgt gctgggtgcg gcggtggcgc ggcatctggc tggtgtgtgt 10680
ggggtgcggg atctgctgtt ggtgagccgg cgtggtccgg atgctccggg tgcggagggt 10740
ctgcgggcgg agctggccgc gttgggggcg gaggtgcgga ttgttgcgtg tgatgtgggg 10800
gagcggcggg aggtggtccg gctgctggag ggtgttcctg ccgggtgtcc gctgacgggt 10860
gtcgtgcatg cggctggtgt gctggacgat gcgacgatcg cctctctcac gcccgagcgg 10920
ctgggcacgg tgttcgcggc caaggtggat gccgctcttt tgctggatga gctgacgcgg 10980
ggtatggagc tgtcggcgtt cgtgctgttc tcctcggccg cggggatcct ggggtcggcc 11040
gggcagggca actacgccgc ggccaatgcc gctctggacg cgctggcgta ccggcggcgg 11100
gcggcgggtc tgccgggggt gtcgctggcg tgggggctgt gggaagaggc cagcgggatg 11160
accgggcacc tggccggcac cgaccaccgg cgcatcatcc gttccggtct gcatcccatg 11220
tcgaccccgg acgcactggc cctcttcgat gcggccctgg ctctggaccg gccggtcctg 11280
ctgcccgccg acctgcgtcc cgccccgccc ctgccgcccc tgctgcagga cctcctgccc 11340
gccacccgcc gccgcaccac ccgcaccacc actaccggtg gtgcggacaa cggcgcccag 11400
ctgcacgccc ggctggccgg ccagacacac gaacaacagc acaccaccct cctcgccctg 11460
gtccgctccc acatcgccac cgtcctgggc cacaccaccc ccgacaccat cccccccgac 11520
cgcgcgttcc gcgacctcgg cttcgactcc ctcaccgccg tcgaactacg caaccggctc 11580
tcccgcacca ccggactccg cctccccacc accctcgcct tcgaccaccc caaccccacc 11640
accctcaccc accacctcca cacacaactc cagccacaac cggacaacgc tgtcgccccc 11700
gtgttggcgg agctcgacaa actcgaatcc gccctctccg ccctcgacaa aaccgacagc 11760
gccagcgaaa gagtcaccct gcggctgaag tcactcatgt tgaggtggaa cgcaccccag 11820
catccgacag ccgaaagcgc tgatgacgac gagaagttca catcggcaac agaggctgag 11880
attttcaaat tcattgacaa cgacctcggc ctgtcctga 11919
<210> 67
<211> 3972
<212> PRT
<213> Artificial Sequence
<220>
<223> type I polyketide synthase AVES 1 (BAA84474.1)
<400> 67
Met Gln Arg Met Asp Gly Gly Glu Glu Pro Arg Pro Ala Ala Gly Glu
1 5 10 15
Val Leu Gly Val Ala Asp Glu Ala Asp Gly Gly Val Val Phe Val Phe
20 25 30
Pro Gly Gln Gly Pro Gln Trp Pro Gly Met Gly Arg Glu Leu Leu Asp
35 40 45
Ala Ser Asp Val Phe Arg Glu Ser Val Arg Ala Cys Glu Ala Ala Phe
50 55 60
Ala Pro Tyr Val Asp Trp Ser Val Glu Gln Val Leu Arg Asp Ser Pro
65 70 75 80
Asp Ala Pro Gly Leu Asp Arg Val Asp Val Val Gln Pro Thr Leu Phe
85 90 95
Ala Val Met Ile Ser Leu Ala Ala Leu Trp Arg Ser Gln Gly Val Glu
100 105 110
Pro Cys Ala Val Leu Gly His Ser Leu Gly Glu Ile Ala Ala Ala His
115 120 125
Val Ser Gly Gly Leu Ser Leu Ala Asp Ala Ala Arg Val Val Thr Leu
130 135 140
Trp Ser Gln Ala Gln Thr Thr Leu Ala Gly Thr Gly Ala Leu Val Ser
145 150 155 160
Val Ala Ala Thr Pro Asp Glu Leu Leu Pro Arg Ile Ala Pro Trp Thr
165 170 175
Glu Asp Asn Pro Ala Arg Leu Ala Val Ala Ala Val Asn Gly Pro Arg
180 185 190
Ser Thr Val Val Ser Gly Ala Arg Glu Ala Val Ala Asp Leu Val Ala
195 200 205
Asp Leu Thr Ala Ala Gln Val Arg Thr Arg Met Ile Pro Val Asp Val
210 215 220
Pro Ala His Ser Pro Leu Met Tyr Ala Ile Glu Glu Arg Val Val Ser
225 230 235 240
Gly Leu Leu Pro Ile Thr Pro Arg Pro Ser Arg Ile Pro Phe His Ser
245 250 255
Ser Val Thr Gly Gly Arg Leu Asp Thr Arg Glu Leu Asp Ala Ala Tyr
260 265 270
Trp Tyr Arg Asn Met Ser Ser Thr Val Arg Phe Glu Pro Ala Ala Arg
275 280 285
Leu Leu Leu Gln Gln Gly Pro Lys Thr Phe Val Glu Met Ser Pro His
290 295 300
Pro Val Leu Thr Met Gly Leu Gln Glu Leu Ala Pro Asp Leu Gly Asp
305 310 315 320
Thr Thr Gly Thr Ala Asp Thr Val Ile Met Gly Thr Leu Arg Arg Gly
325 330 335
Gln Gly Thr Leu Asp His Phe Leu Thr Ser Leu Ala Gln Leu Arg Gly
340 345 350
His Gly Glu Thr Ser Ala Thr Thr Val Leu Ser Ala Arg Leu Thr Ala
355 360 365
Leu Ser Pro Thr Gln Gln Gln Ser Leu Leu Leu Asp Leu Val Arg Ala
370 375 380
His Thr Met Ala Val Leu Asn Asp Asp Gly Asn Glu Arg Thr Ala Ser
385 390 395 400
Asp Ala Gly Pro Ser Ala Ser Phe Ala His Leu Gly Phe Asp Ser Val
405 410 415
Met Gly Val Glu Leu Arg Asn Arg Leu Ser Lys Ala Thr Gly Leu Arg
420 425 430
Leu Pro Val Thr Leu Ile Phe Asp His Thr Thr Pro Ala Ala Val Ala
435 440 445
Ala Arg Leu Arg Thr Ala Ala Leu Gly His Leu Asp Glu Asp Thr Ala
450 455 460
Pro Val Pro Asp Ser Pro Ser Gly His Gly Gly Thr Ala Ala Ala Asp
465 470 475 480
Asp Pro Ile Ala Ile Ile Gly Met Ala Cys Arg Phe Pro Gly Gly Val
485 490 495
Arg Ser Pro Lys Asp Leu Trp Glu Leu Ala Ala Ser Gly Gly Asp Ala
500 505 510
Ile Gly Pro Phe Pro Thr Asp Arg Gly Trp Pro Thr Glu Gln Arg His
515 520 525
Ala Gln Asp Pro Thr Gln Pro Gly Thr Phe Tyr Pro Gln Gly Gly Gly
530 535 540
Phe Leu His Asp Ala Ala His Phe Asp Ala Gly Phe Phe Gly Ile Ser
545 550 555 560
Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu
565 570 575
Thr Ser Trp Glu Ala Phe Glu Arg Ala Gly Ile Asp Pro Leu Ser Val
580 585 590
Arg Gly Ser Arg Thr Gly Val Phe Ala Gly Ala Leu Ser Phe Asp Tyr
595 600 605
Gly Pro Arg Met Asp Thr Ala Ser Ser Glu Gly Ala Ala Asp Val Glu
610 615 620
Gly His Ile Leu Thr Gly Thr Thr Gly Ser Val Leu Ser Gly Arg Ile
625 630 635 640
Ala Tyr Ser Phe Gly Leu Glu Gly Pro Ala Ile Thr Val Asp Thr Gly
645 650 655
Cys Ser Ala Ser Leu Val Thr Leu His Leu Ala Cys Gln Ser Leu Arg
660 665 670
Ser Gly Glu Cys Thr Leu Ala Leu Ala Gly Gly Val Ser Val Met Ser
675 680 685
Thr Leu Gly Met Phe Ile Glu Phe Ser Arg Gln Arg Gly Leu Ser Val
690 695 700
Asp Gly Arg Cys Lys Ala Tyr Ser Ala Ala Ala Asp Gly Thr Gly Trp
705 710 715 720
Gly Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala Val
725 730 735
Arg Leu Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn
740 745 750
Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala Gln
755 760 765
Glu Arg Val Ile Arg Gln Ala Leu Ala Asn Ala Gly Leu Ser Val Ala
770 775 780
Asp Val Asp Val Val Glu Gly His Gly Thr Gly Thr Thr Leu Gly Asp
785 790 795 800
Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Arg Ala Gly
805 810 815
Asp Arg Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly His Thr
820 825 830
Met Ala Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Met Ala Leu
835 840 845
Arg Glu Gly Val Leu Pro Arg Thr Leu His Val Asp Lys Pro Ser Pro
850 855 860
Gln Val Asp Trp Ser Ala Gly Ala Val Arg Leu Leu Thr Glu Ala Val
865 870 875 880
Pro Trp Pro Gly Asp Ala Ala Gly Arg Leu Arg Arg Ala Gly Val Ser
885 890 895
Ser Phe Gly Ile Gly Gly Thr Asn Ala His Val Ile Leu Glu Glu Ala
900 905 910
Pro Ala Ala Gly Gly Cys Val Ala Gly Gly Gly Val Leu Glu Gly Ala
915 920 925
Pro Gly Leu Ala Ile Ser Val Ala Glu Ser Val Ala Ala Pro Val Ala
930 935 940
Val Ser Ala Pro Val Ala Glu Ser Val Pro Val Pro Val Pro Val Pro
945 950 955 960
Val Pro Val Pro Val Ser Ala Arg Ser Glu Ala Gly Leu Arg Ala Gln
965 970 975
Ala Glu Ala Leu Arg Gln Tyr Val Ala Val Arg Pro Asp Val Ser Leu
980 985 990
Ala Asp Val Gly Ala Gly Leu Ala Cys Gly Arg Ala Val Leu Glu His
995 1000 1005
Arg Ala Val Val Leu Ala Ala Asp Arg Glu Glu Leu Val Gln Gly Leu
1010 1015 1020
Gly Ala Leu Ala Ala Gly Glu Pro Asp Arg Arg Val Thr Thr Gly His
1025 1030 1035 1040
Ala Pro Gly Gly Asp Arg Gly Gly Val Val Phe Val Phe Pro Gly Gln
1045 1050 1055
Gly Gly Gln Trp Ala Gly Met Gly Val Arg Leu Leu Ala Ser Ser Pro
1060 1065 1070
Val Phe Ala Arg Arg Met Gln Ala Cys Glu Glu Ala Leu Ala Pro Trp
1075 1080 1085
Val Asp Trp Ser Val Val Asp Ile Leu Arg Arg Asp Ala Gly Asp Ala
1090 1095 1100
Val Trp Glu Arg Ala Asp Val Val Gln Pro Val Leu Phe Ser Val Met
1105 1110 1115 1120
Val Ser Leu Ala Ala Leu Trp Arg Ser Tyr Gly Ile Glu Pro Asp Ala
1125 1130 1135
Val Leu Gly His Ser Gln Gly Glu Ile Ala Ala Ala His Val Cys Gly
1140 1145 1150
Ala Leu Ser Leu Lys Asp Ala Ala Lys Thr Val Ala Leu Arg Ser Arg
1155 1160 1165
Ala Leu Ala Ala Val Arg Gly Arg Gly Gly Met Ala Ser Val Pro Leu
1170 1175 1180
Pro Ala Gln Glu Val Glu Gln Leu Ile Gly Glu Arg Trp Ala Gly Arg
1185 1190 1195 1200
Leu Trp Val Ala Ala Val Asn Gly Pro Arg Ser Thr Ala Val Ser Gly
1205 1210 1215
Asp Ala Glu Ala Val Asp Glu Val Leu Ala Tyr Cys Ala Gly Thr Gly
1220 1225 1230
Val Arg Ala Arg Arg Ile Pro Val Asp Tyr Ala Ser His Cys Pro His
1235 1240 1245
Val Gln Pro Leu Arg Glu Glu Leu Leu Glu Leu Leu Gly Asp Ile Ser
1250 1255 1260
Pro Gln Pro Ser Gly Val Pro Phe Phe Ser Thr Val Glu Gly Thr Trp
1265 1270 1275 1280
Leu Asp Thr Thr Thr Leu Asp Ala Ala Tyr Trp Tyr Arg Asn Leu His
1285 1290 1295
Gln Pro Val Arg Phe Ser Asp Ala Val Gln Ala Leu Ala Asp Asp Gly
1300 1305 1310
His Arg Val Phe Val Glu Val Ser Pro His Pro Thr Leu Val Pro Ala
1315 1320 1325
Ile Glu Asp Thr Thr Glu Asp Thr Ala Glu Asp Val Thr Ala Ile Gly
1330 1335 1340
Ser Leu Arg Arg Gly Asp Asn Asp Thr Arg Arg Phe Leu Thr Ala Leu
1345 1350 1355 1360
Ala His Thr His Thr Thr Gly Ile Gly Thr Pro Thr Thr Trp His His
1365 1370 1375
His Tyr Thr His His His Thr His Pro His Pro His Thr His Leu Asp
1380 1385 1390
Leu Pro Thr Tyr Pro Phe Gln His Gln His Tyr Trp Leu Glu Ser Ser
1395 1400 1405
Gln Pro Gly Ala Gly Ser Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala
1410 1415 1420
Gly Ser Gly Arg Ala Gly Thr Ala Gly Gly Thr Ala Glu Val Glu Ser
1425 1430 1435 1440
Arg Phe Trp Asp Ala Val Ala Arg Gln Asp Leu Glu Thr Val Ala Thr
1445 1450 1455
Thr Leu Ala Val Pro Pro Ser Ala Gly Leu Asp Thr Val Val Pro Ala
1460 1465 1470
Leu Ser Ala Trp His Arg His Gln His Asp Gln Ala Arg Ile Asn Thr
1475 1480 1485
Trp Thr Tyr Gln Glu Thr Trp Lys Pro Leu Thr Leu Pro Thr Thr His
1490 1495 1500
Gln Pro His Gln Thr Trp Leu Ile Ala Ile Pro Glu Thr Gln Thr His
1505 1510 1515 1520
His Pro His Ile Thr Asn Ile Leu Thr Asn Leu His His His Gly Ile
1525 1530 1535
Thr Pro Ile Pro Leu Thr Leu Asn His Thr His Thr Asn Pro Gln His
1540 1545 1550
Leu His His Thr Leu His His Thr Arg Gln Gln Ala Gln Asn His Thr
1555 1560 1565
Thr Gly Ala Ile Thr Gly Leu Leu Ser Leu Leu Ala Leu Asp Glu Thr
1570 1575 1580
Pro His Pro His His Pro His Thr Pro Thr Gly Thr Leu Leu Asn Leu
1585 1590 1595 1600
Thr Leu Thr Gln Thr His Thr Gln Thr His Pro Pro Thr Pro Leu Trp
1605 1610 1615
Tyr Ala Thr Thr Asn Ala Thr Thr Thr His Pro Asn Asp Pro Leu Thr
1620 1625 1630
His Pro Thr Gln Ala Gln Thr Trp Gly Leu Ala Arg Thr Thr Leu Leu
1635 1640 1645
Glu His Pro Thr His Thr Ala Gly Ile Ile Asp Leu Pro Thr Thr Pro
1650 1655 1660
Thr Pro His Thr Leu Gln His Leu Thr Gln Thr Leu Thr Gln Pro His
1665 1670 1675 1680
His Gln Thr Gln Leu Ala Ile Arg Thr Thr Gly Thr His Thr Arg Arg
1685 1690 1695
Leu Thr Pro Thr Thr Leu Thr Pro Thr His Gln Pro Pro Thr Pro Thr
1700 1705 1710
Pro His Gly Thr Thr Leu Ile Thr Gly Gly Thr Gly Ala Leu Ala Thr
1715 1720 1725
His Leu Thr His His Leu Thr Thr His Gln Pro Thr Gln His Leu Leu
1730 1735 1740
Leu Thr Ser Arg Thr Gly Pro His Thr Pro His Ala Gln His Leu Thr
1745 1750 1755 1760
Thr Gln Leu Gln Gln Lys Gly Ile His Leu Thr Ile Thr Thr Cys Asp
1765 1770 1775
Thr Ser Asn Pro Asp Gln Leu Gln Gln Leu Leu Asn Thr Ile Pro Pro
1780 1785 1790
Gln His Pro Leu Thr Thr Val Ile His Thr Ala Gly Ile Leu Asp Asp
1795 1800 1805
Ala Thr Leu Thr Asn Leu Thr Pro Thr Gln Leu Asn Asn Val Leu Arg
1810 1815 1820
Ala Lys Ala His Ser Ala His Leu Leu His Gln Leu Thr Gln His Thr
1825 1830 1835 1840
Pro Leu Thr Ala Phe Val Leu Tyr Ser Ser Ala Ala Ala Thr Phe Gly
1845 1850 1855
Ala Pro Gly Gln Ala Asn Tyr Ala Ala Ala Asn Ala Tyr Leu Asp Ala
1860 1865 1870
Leu Ala His His Arg His Thr His His Leu Pro Ala Thr Ser Ile Ala
1875 1880 1885
Trp Gly Thr Trp Gln Gly Asn Gly Leu Ala Asp Ser Asp Lys Ala Arg
1890 1895 1900
Ala Tyr Leu Asp Arg Arg Gly Phe Arg Pro Met Ser Pro Glu Leu Ala
1905 1910 1915 1920
Thr Ala Ala Val Thr Gln Ala Ile Ala Asp Thr Glu Arg Pro Tyr Val
1925 1930 1935
Val Ile Ala Asp Ile Asp Trp Ser Lys Ile Glu His Thr Ser Gln Thr
1940 1945 1950
Ser Asp Leu Val Ser Ala Ala Arg Glu Arg Glu Pro Ala Val Gln Arg
1955 1960 1965
Pro Thr Pro Pro Ala Glu Leu His Lys Thr Leu Ala His Gln Thr Ser
1970 1975 1980
Ala Asp Gln Arg Ala Ala Leu Leu Glu Leu Val Arg Asp His Val Ala
1985 1990 1995 2000
Ala Val Leu Arg His Ala Asp Pro Lys Ala Ile Ala Pro Asp Gln Ser
2005 2010 2015
Phe Arg Ala Leu Gly Phe Asp Ser Leu Thr Ala Val Glu Phe Arg Asn
2020 2025 2030
Leu Leu Ile Lys Ala Thr Gly Leu Arg Leu Pro Val Ser Leu Val Phe
2035 2040 2045
Asp His Pro Thr Pro Ala Lys Leu Ala Val His Leu Gln Asn Gln Leu
2050 2055 2060
Arg Gly Thr Ala Ala Glu Ser Ala Pro Ser Ala Ala Ala Val Thr Ala
2065 2070 2075 2080
Glu Ala Ser Val Thr Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg
2085 2090 2095
Phe Pro Gly Gly Val Thr Ser Ala Asp Asp Phe Trp Asp Leu Ile Ser
2100 2105 2110
Ser Glu Gln Asp Ala Ile Gly Gly Phe Pro Thr Asp Arg Gly Trp Asp
2115 2120 2125
Leu Asp Thr Leu Tyr Asp Pro Asp Pro Asp His Pro Gly Thr Cys Tyr
2130 2135 2140
Thr Arg Asn Gly Gly Phe Leu Tyr Asp Ala Gly His Phe Asp Ala Glu
2145 2150 2155 2160
Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln
2165 2170 2175
Arg Leu Leu Leu Glu Thr Ala Trp Glu Thr Ile Glu His Ala Gly Ile
2180 2185 2190
Asn Pro His Thr Leu His Gly Thr Pro Thr Gly Val Phe Thr Gly Thr
2195 2200 2205
Asn Gly Gln Asp Tyr Ala Leu Arg Val His Asn Ala Gly Gln Ser Thr
2210 2215 2220
Asp Gly Phe Ala Leu Thr Gly Thr Ala Gly Ser Val Ile Ser Gly Arg
2225 2230 2235 2240
Ile Ser Tyr Thr Phe Gly Phe Glu Gly Pro Ala Val Ser Val Asp Thr
2245 2250 2255
Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu
2260 2265 2270
Arg Ala Gly Glu Cys Ser Met Ala Leu Ala Gly Gly Val Thr Val Met
2275 2280 2285
Ser Ser Pro Gly Ala Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ala
2290 2295 2300
Ala Asp Gly His Cys Lys Ala Phe Ser Ala Ala Ala Asp Gly Thr Gly
2305 2310 2315 2320
Trp Gly Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala
2325 2330 2335
His Arg Asn Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val
2340 2345 2350
Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser
2355 2360 2365
Gln Gln Arg Val Ile Arg Gln Ala Leu Ala Asn Ala Gly Leu Ser Ala
2370 2375 2380
Gly Asp Val Asp Ala Val Glu Ala His Gly Thr Gly Thr Thr Leu Gly
2385 2390 2395 2400
Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Asp Arg
2405 2410 2415
Ala Gly Glu Gly Pro Leu Trp Leu Gly Ser Val Lys Ser Asn Val Gly
2420 2425 2430
His Thr Gln Ala Ala Ala Gly Val Ala Gly Val Ile Lys Met Val Met
2435 2440 2445
Ala Leu Arg His Gly Leu Leu Pro Arg Thr Leu His Val Asp Glu Pro
2450 2455 2460
Ser Pro His Val Asp Trp Ser Ala Gly Ala Val Gln Leu Leu Thr Glu
2465 2470 2475 2480
Thr Val Pro Trp Pro Gly Gly Glu Gly Arg Leu Arg Arg Ala Gly Val
2485 2490 2495
Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Glu
2500 2505 2510
Ala Pro Ala Asp Asp Val Pro Gly Gly Pro Pro Ala Gly Glu Gly Asp
2515 2520 2525
Ala Gly Ser Asp Asp Glu Ala Ala Ala Gly Ser Pro Gly Val Trp Pro
2530 2535 2540
Trp Leu Val Ser Ala Lys Ser Gln Pro Ala Leu Arg Ala Gln Ala Gln
2545 2550 2555 2560
Ala Leu His Ala His Leu Thr Asp His Pro Gly Leu Asp Leu Ala Asp
2565 2570 2575
Val Gly Tyr Thr Leu Ala His Ala Arg Ala Val Phe Asp His Arg Ala
2580 2585 2590
Thr Leu Ile Ala Ala Asp Arg Asp Thr Phe Leu Gln Ala Leu Gln Ala
2595 2600 2605
Leu Ala Ala Gly Glu Pro His Pro Ala Val Ile His Ser Ser Ala Pro
2610 2615 2620
Gly Gly Thr Gly Thr Gly Glu Ala Ala Gly Lys Thr Ala Phe Ile Cys
2625 2630 2635 2640
Ser Gly Gln Gly Thr Gln Arg Pro Gly Met Ala His Gly Leu Tyr His
2645 2650 2655
Thr His Pro Val Phe Ala Ala Ala Leu Asn Asp Ile Cys Thr His Leu
2660 2665 2670
Asp Pro His Leu Asp His Pro Leu Leu Pro Leu Leu Thr Gln Asn Asp
2675 2680 2685
Asn Asp Asn Glu Asp Ala Ala Ala Leu Leu Gln Gln Thr Arg Tyr Ala
2690 2695 2700
Gln Pro Ala Leu Phe Ala Phe Gln Val Ala Leu His Arg Leu Leu Thr
2705 2710 2715 2720
Asp Gly Tyr His Ile Thr Pro His Tyr Tyr Ala Gly His Ser Leu Gly
2725 2730 2735
Glu Ile Thr Ala Ala His Leu Ala Gly Ile Leu Thr Leu Thr Asp Ala
2740 2745 2750
Thr Thr Leu Ile Thr Gln Arg Ala Thr Leu Met Gln Thr Met Pro Pro
2755 2760 2765
Gly Thr Met Thr Thr Leu His Thr Thr Pro His His Ile Thr His His
2770 2775 2780
Leu Thr Ala His Glu Asn Asp Leu Ala Ile Ala Ala Ile Asn Thr Pro
2785 2790 2795 2800
Thr Ser Leu Val Ile Ser Gly Thr Pro His Thr Val Gln His Ile Thr
2805 2810 2815
Thr Leu Cys Gln Gln Gln Gly Ile Lys Thr Lys Thr Leu Pro Thr Asn
2820 2825 2830
His Ala Phe His Ser Pro His Thr Asn Pro Ile Leu Asn Gln Leu His
2835 2840 2845
Gln His Thr Gln Thr Leu Thr Tyr His Pro Pro His Thr Pro Leu Ile
2850 2855 2860
Thr Ala Asn Thr Pro Pro Asp Gln Leu Leu Thr Pro His Tyr Trp Thr
2865 2870 2875 2880
Gln Gln Ala Arg Asn Thr Val Asp Tyr Ala Thr Thr Thr Gln Thr Leu
2885 2890 2895
His Gln His Gly Val Thr Thr Tyr Ile Glu Leu Gly Pro Asp Asn Thr
2900 2905 2910
Leu Thr Thr Leu Thr His His Asn Leu Pro Asn Pro Pro Thr Thr Thr
2915 2920 2925
Leu Thr Leu Thr His Pro His His His Pro Gln Thr His Leu Leu Thr
2930 2935 2940
Asn Leu Ala Lys Thr Thr Thr Thr Trp His Pro His His Tyr Thr His
2945 2950 2955 2960
His Asp Asn Gln Pro His Thr His Thr His Leu Asp Leu Pro Thr Tyr
2965 2970 2975
Pro Phe Gln His His His Tyr Trp Leu Glu Ser Thr Gln Pro Gly Ala
2980 2985 2990
Gly Asn Val Ser Ala Ala Gly Leu Asp Pro Thr Glu His Pro Leu Leu
2995 3000 3005
Gly Ala Thr Leu Glu Leu Ala Thr Asp Gly Gly Ala Leu Leu Ala Gly
3010 3015 3020
Arg Leu Ser Leu Arg Ser His Pro Trp Leu Ala Asp His Ala Val Gly
3025 3030 3035 3040
Gly Thr Val Leu Leu Ser Gly Ala Thr Phe Leu Glu Leu Ala Leu His
3045 3050 3055
Ala Gly Thr Tyr Val Gly Cys Asp Arg Val Asp Glu Leu Thr Leu His
3060 3065 3070
Ala Pro Leu Val Val Pro Val Asp Gly Gly Val Ser Val Gln Val Gly
3075 3080 3085
Val Ala Ala Ala Asp Gly Glu Gly Arg Arg Leu Val Ser Val Tyr Ala
3090 3095 3100
Arg Gly Gly Ser Ala Cys Gly Gly Gly Gly Ala Ser Gly Gly Val Trp
3105 3110 3115 3120
Thr Cys His Ala Ser Gly Val Leu Val Glu Ala Ala Ala Gly Gly Val
3125 3130 3135
Val Val Asp Gly Leu Ala Gly Val Trp Pro Pro Arg Gly Ala Val Ala
3140 3145 3150
Val Asp Val Asp Gly Val Arg Asp Arg Leu Ala Gly Ala Gly Cys Val
3155 3160 3165
Leu Gly Pro Val Phe Ser Gly Leu Arg Ala Val Trp Arg Asp Gly Gly
3170 3175 3180
Asp Leu Leu Ala Glu Val Cys Leu Pro Glu Glu Ala Trp Gly Asp Ala
3185 3190 3195 3200
Ala Gly Phe Gly Leu His Pro Ala Leu Leu Asp Gly Val Val Gln Pro
3205 3210 3215
Leu Ser Val Leu Leu Pro Gly Gly Thr Gly Phe Gly Glu Gly Ala Gly
3220 3225 3230
Phe Gly Glu Gly Val Arg Val Pro Ala Val Trp Gly Gly Val Ser Leu
3235 3240 3245
His Arg Ala Gly Val Thr Gly Val Arg Val Arg Val Ser Ala Val Gly
3250 3255 3260
Arg Gly Gly Gly Arg Glu Ala Val Ser Val Val Val Gly Asp Glu Ala
3265 3270 3275 3280
Gly Val Pro Val Ala Ser Val Asp Arg Leu Glu Leu Arg Pro Val Asp
3285 3290 3295
Met Gly Gln Leu Arg Ala Val Ser Val Ser Ala Gly Arg Arg Gly Ser
3300 3305 3310
Leu Tyr Ala Val Gln Trp Ala Glu Val Gly Pro Val Pro Val Cys Gly
3315 3320 3325
Gln Ala Trp Ala Trp His Glu Asp Val Gly Glu Ser Gly Gly Gly Pro
3330 3335 3340
Val Pro Gly Val Val Val Leu Arg Cys Pro Asp Ala Gly Ala Gly Gly
3345 3350 3355 3360
Gly Gly Gly Gly Gly Gly Gly Gly Gly Val Gly Glu Val Val Gly Gly
3365 3370 3375
Val Leu Gly Val Val Gln Gly Trp Leu Gly Leu Glu Arg Phe Ala Gly
3380 3385 3390
Ser Arg Leu Val Val Val Thr Arg Gly Ala Val Val Ala Gly Pro Glu
3395 3400 3405
Asp Gly Pro Val Asp Val Val Gly Ala Ser Val Trp Gly Leu Val Arg
3410 3415 3420
Ser Ala Gln Ala Glu His Pro Asp Arg Phe Val Leu Leu Asp Leu Asp
3425 3430 3435 3440
Thr Asp Thr Gly Thr Asp Leu Asp Thr Gly Ala Gly Ala Gly Trp Gly
3445 3450 3455
Val Asp Gly Gly Arg Val Ala Ala Val Val Ala Cys Gly Glu Pro Gln
3460 3465 3470
Leu Ala Val Arg Gly Glu Arg Leu Leu Ala Ala Arg Leu Lys Arg Leu
3475 3480 3485
Glu Ser Ser Gly Asp Val Pro Ala Gln Arg Ser Gly Asp Thr Arg Ala
3490 3495 3500
Arg Arg Ser Asp Val Pro Ala Gln Arg Ser Gly Gly Val Pro Ala Arg
3505 3510 3515 3520
Arg Ser Val Asp Val Ser Gly Arg Glu Val Leu Pro Trp Leu Ser Gly
3525 3530 3535
Gly Ser Val Leu Val Thr Gly Gly Thr Gly Val Leu Gly Ala Ala Val
3540 3545 3550
Ala Arg His Leu Ala Gly Val Cys Gly Val Arg Asp Leu Leu Leu Val
3555 3560 3565
Ser Arg Arg Gly Pro Asp Ala Pro Gly Ala Glu Gly Leu Arg Ala Glu
3570 3575 3580
Leu Ala Ala Leu Gly Ala Glu Val Arg Ile Val Ala Cys Asp Val Gly
3585 3590 3595 3600
Glu Arg Arg Glu Val Val Arg Leu Leu Glu Gly Val Pro Ala Gly Cys
3605 3610 3615
Pro Leu Thr Gly Val Val His Ala Ala Gly Val Leu Asp Asp Ala Thr
3620 3625 3630
Ile Ala Ser Leu Thr Pro Glu Arg Leu Gly Thr Val Phe Ala Ala Lys
3635 3640 3645
Val Asp Ala Ala Leu Leu Leu Asp Glu Leu Thr Arg Gly Met Glu Leu
3650 3655 3660
Ser Ala Phe Val Leu Phe Ser Ser Ala Ala Gly Ile Leu Gly Ser Ala
3665 3670 3675 3680
Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu Ala
3685 3690 3695
Tyr Arg Arg Arg Ala Ala Gly Leu Pro Gly Val Ser Leu Ala Trp Gly
3700 3705 3710
Leu Trp Glu Glu Ala Ser Gly Met Thr Gly His Leu Ala Gly Thr Asp
3715 3720 3725
His Arg Arg Ile Ile Arg Ser Gly Leu His Pro Met Ser Thr Pro Asp
3730 3735 3740
Ala Leu Ala Leu Phe Asp Ala Ala Leu Ala Leu Asp Arg Pro Val Leu
3745 3750 3755 3760
Leu Pro Ala Asp Leu Arg Pro Ala Pro Pro Leu Pro Pro Leu Leu Gln
3765 3770 3775
Asp Leu Leu Pro Ala Thr Arg Arg Arg Thr Thr Arg Thr Thr Thr Thr
3780 3785 3790
Gly Gly Ala Asp Asn Gly Ala Gln Leu His Ala Arg Leu Ala Gly Gln
3795 3800 3805
Thr His Glu Gln Gln His Thr Thr Leu Leu Ala Leu Val Arg Ser His
3810 3815 3820
Ile Ala Thr Val Leu Gly His Thr Thr Pro Asp Thr Ile Pro Pro Asp
3825 3830 3835 3840
Arg Ala Phe Arg Asp Leu Gly Phe Asp Ser Leu Thr Ala Val Glu Leu
3845 3850 3855
Arg Asn Arg Leu Ser Arg Thr Thr Gly Leu Arg Leu Pro Thr Thr Leu
3860 3865 3870
Ala Phe Asp His Pro Asn Pro Thr Thr Leu Thr His His Leu His Thr
3875 3880 3885
Gln Leu Gln Pro Gln Pro Asp Asn Ala Val Ala Pro Val Leu Ala Glu
3890 3895 3900
Leu Asp Lys Leu Glu Ser Ala Leu Ser Ala Leu Asp Lys Thr Asp Ser
3905 3910 3915 3920
Ala Ser Glu Arg Val Thr Leu Arg Leu Lys Ser Leu Met Leu Arg Trp
3925 3930 3935
Asn Ala Pro Gln His Pro Thr Ala Glu Ser Ala Asp Asp Asp Glu Lys
3940 3945 3950
Phe Thr Ser Ala Thr Glu Ala Glu Ile Phe Lys Phe Ile Asp Asn Asp
3955 3960 3965
Leu Gly Leu Ser
3970
<210> 68
<211> 13032
<212> DNA
<213> Artificial Sequence
<220>
<223> milA1 gene of S. bingchenggensis BCW-1 (Accession no. CP002047)
<400> 68
ttgcccaaag cccagaacga gttcgcagtg gccggtcatc cgtggatcct ctccgggcac 60
accggaaccg cgctgcgggc ccaggcacgc cggctccacg accatgtcgc cgaccaccct 120
cggctccgtc cggaagacat cgcccacacg ctggcgagca gcggcccggc gctcacccat 180
cgcgcggcgg tgatcgcggc ggaccgggaa ggacatctcc gggggctcga cgcggtggcc 240
cggggtgagg acacccccgg tgtcgtacgg ggcacggcgg ccgcgggcgg cgacggggtc 300
gcgttcgtct tccccggcca gggcacccag tggcccggta tggccgccga tctgctgacg 360
gtctcccccg ccttcagccg ggcggtcgac gcctgcgccg aggcgttcga accgtatgtc 420
tcctggtcac cggaggccgt gctgcggggc gctccgggcg cgccgcccct ggaggggacc 480
gatgtggtgc agccgacgct gttcgccgtc atggtggggc tggccgagct gtggcggact 540
cttggggtga gcccgacgtc gatcgtgggc cactgcatcg gggagatcgc ggcagcccat 600
ctctgcggcg ccctgtcgct gtccgacgcg gcgcgcgtgg tgatcgagag cagccgggcc 660
caggcgacgc tctccgggtc gggtgcgctg atcgcggtcg cgcggtccga ggcgcagctg 720
cttccgttgc tgcggcggtg gccgggcagg ctgacgatcg ccgcggtcaa cggcccgatg 780
gccacggtcg tctccggcga tcggccggcc gccgacgagc tgttggcgga gttcgcccgt 840
gccggtgtcc gggcccgcga ggtggcgatc gacatccccg cgcactcgcc gttcatggcc 900
cccctcaggg acggtctgct cgactcgctg tcatcggtca ccgcgggtgc gtcgcggctg 960
ccgttccact cctcggtcat cggggggccg ctggagaccc aagggctcga cgcggcttac 1020
tggtaccgga acctcgccga cacggtccgc ttcgaaagcg tcgtcacggg gctgctgcgg 1080
cagggcacac gctgcttcgt ggagctgagc ccgcacccga tgctgaccat gtgtgtgcag 1140
gccaccgccg aggaggtggt cggcggtgag cgcgtcgtga tcctgccgac gctgcatcgc 1200
gggcaggccg ccgtcgagtc cgttcgcacc acgctggccg agctgtacgt acggggcgca 1260
ctggatgacc atcgggcggc gttctcggtg ccgggcggcc gcctgatcac cctgcctctc 1320
gagccgcccg cggacacgtc cgtagagctc gccgacgccc cggacccggc ggaggcctgc 1380
cggcccccct tggtggagcg gcttgcccgg ctctccaccg cggagcggaa gcggcggctg 1440
cgcgagctgg tgggcgtcga ggcggccaag gtcctcgagg acgtcgccgg ggcggacgcg 1500
ccgggccacg gcatcgcgga gcaggagcac ttcgtcactt cgggcttcga ctccgcggcc 1560
gcggtcgcgc tgcgcaaccg cctgaacgac gccaccggtt tgctgctgcc cttcaccctg 1620
gccttcgacc atccgacacc cgccgccgtc gccgaccatc tgcactcccg gctcttcgat 1680
caccagggcg gcgggcagcc gggcgccgac ggccggcccg accccgcggc ggcggccggt 1740
ccggccaggg ccgacgacga gccgatcgcc gtcatcggca tggcgggccg cttccccggg 1800
ggcgcccgta ccccggagga gctgtgggaa ctggtcgccg aaggcaccga cgccctctcg 1860
cccttcccgg agggccgggg ctgggatccg ctgcggctct acgatccgga ccccgcccgg 1920
cccggcacgt actaccagcg cgaagcggga ttcctccacg acgccgacaa gttcgacgcc 1980
gagttcttcg gcatcgcgcc acgcgaggcc accgcaatgg atccccagca gcggctgctc 2040
ctggagacct cctgggaggc gctcgaacgg gcgcggatcg acccgaccgc gctgcgcggc 2100
agccgcaccg gggtgttcgt cggcgtggcc ccgctggact acagcccccg aatgcaccag 2160
gcgtcgccgg agctggaggg ccatctgctg accggcaaca tcggcgccgc ggcctcgggg 2220
cggatctcct acgtactcgg gcttgagggg cccgcggtgt ccgtggacac ggcgtgctcg 2280
tcgtccctgg tcgccctgca tctggcggcc caggcgctgc gggccgggga gtgctcgctg 2340
gccctggtcg gcggggcgac ggtcctctcg acccccggca tgttcatcga gttctcgcgg 2400
cagcgcggtc tggctccgga cggccgctgc aaggcgtacg cggccgccgc ggacggcacc 2460
ggctggtccg agggtgtggg catgctgctc gtcgagcggc tgtccgacgc gcgacggctc 2520
ggacaccagg tgcttgcggt ggtacggggc tccgccgtca accaggacgg ggcgagcaac 2580
ggcttcacgg cgcccagcgg tccatcacag caacaggtca tccgggcggc cctggccaat 2640
gccggggtgt cggctccgga ggtcgacgcg gtggaggggc acggcaccgg cacccggttg 2700
ggcgatccga tcgaggcgca ggcgctgctg gccgcctacg ggcaggggcg ggcggccgac 2760
cggccgctgt ggctggggtc gatcaagtcg aacatcggac acacccagtg ggccgcgggt 2820
gtcatcgggg tcatcaaaat ggtgctcgcg ctccagcacg gtgtgctgcc gcgcacgctg 2880
cacgtggaca agccgtcgga ttacgtggac tggtcggccg gggccgtacg gctgttgacg 2940
gagccggtgc cctggccgga gcggggccac ccgcgccggg cgggggtgtc gtccttcggg 3000
gtgagcggca ccaacgccca tgtcatcctc gagcaggcaa cgccatcgtc cacggtggct 3060
cccggggggc ataccgccga ggccgggcct cccctgccgt gggtggtctc ggcgaagacg 3120
ccccaggcac tgcgcgacca ggcccgccgc ctgcacgaac acctcaccgc ccagccacag 3180
ctccaaccgg ccgacgtcgg ccacaccctc gccaccggcc gcgccacctt cgaccaccgg 3240
gccgtcctca tcggctccga ccgcgaacaa ctcctccacg gcctggacgc gctcgccacc 3300
ggccggcccg acccagcggt ccaccagacg tcggaccgtc ccgccaccgc cgacggccgt 3360
atcgtcttcg tcttccccgg acaaggcggt caatgggcgg gcatgggcct acggctgctg 3420
aacgcctcac ccgtcttcac cgagcggatg gccgcctgcg aacaggccct ctccccctac 3480
gtcgactggt cactcacgga catcctccac cggccggccg acgacgccgt atggcaacgc 3540
gccgacatcg tccagcccgc cctgttctcg atcatggtgt ccctggccgc gctctggcgc 3600
tcttgcggca tcgaaccgga cgccgtcctc ggccactccc aaggcgagat cgccgcggcc 3660
cacgtctgcg gcgccctgac gctccacgac gcggccaagg tcatcgccct gcgcagccag 3720
gccctccaag ccgtacgcgg cgccgggggc atggcctccg tacccctgcc cgcggaccag 3780
gtcaccgagg atctgcgcac ccactggccc gaccggctat gggtggccgc caccaactcc 3840
cccacggcaa ccgtcatctc gggaaacacc gacgcgcttg acgaagcgct cgaccactac 3900
cacgcccacg acgtacgggc caagcgcatc ccggtcgact acgcctccca ctgcccccat 3960
atcgacgcgg tggccgagcg actgcccgac ctgctgggcg gcatcgtccc gcgcgccgcc 4020
gacatcccct tctactccac ggttgacggc cgatgggccg agccgaccga gctcgacgcc 4080
gactactggt accgcaacct ccgcagcccg gtacggttcg cccacgccgt ccacgccctc 4140
accgagaccg accaccgcac ctttgtcgaa gtcagcccac accccacgct cacccccgcc 4200
atcacggcca ccaccgaaac caccgaccgc accaccaccg tcatcgcctc gctccaccgc 4260
gaccacgacg acacccacca catcctcacc aacctcgccc aggcccacat ccacggccac 4320
accatcgact ggcgacacca ctaccagact ctgcgcccca ccccacccca tatcgacctc 4380
cccacctacc ccttccaaca ccaccactac tggctccacg actccaccga ggacaaggcg 4440
gtgggtacgg acctcgccgc ggcccgcttc tgggaggcgg tccacggcga ggacaccaac 4500
gccgtcgccg cgctcctcga cgtcgagccg ggcacctcac tggacgcgct gctgccggcc 4560
ctgtccgcct ggcacggtcg gcgtcgcgac caggccatca ccgacacctg gtgttaccgc 4620
gacatctgga agccggccga cctcaccgcc gcgcgccccc ggccgtccgg ccgatggctt 4680
gtcgcgatct ccgcagggcg ggccgatcac ctccacgtca gtgccgtcct ggacgctctg 4740
gaacgccagg gtctgcccat cgccaccctc gtcctcgacg acacccacac cgaactcccc 4800
ctgctggagc ggcatctcgc acaggcgatc gcgagcgatg ggccggccat cggcggcgtg 4860
ctctcgctgc tcgccctcga cgaggggcca catccgcgcc acccggaggt gcccgtcggc 4920
accgccctca ccctcagcct gatccaggcg ctcatcgcac gcgaggacat ggcgccccgg 4980
ctgtggctgg ccacccacga ggccgtcgcc acctcgtccg cggatacgct cgatcacccc 5040
ctccaggcga tggtctgggg gctgggacgc accgccgcac tcgaacaccc cgatctgtgg 5100
ggcggcctca tcgaccttcc ggacactctc accgaacggg tcctccacgg cctcgtcacg 5160
gcgctgacca cctgtcacga cgaggacgaa ctcgcgctgc gcgccaccgg cccacgcacc 5220
cggcgcctga tccggacgcc gtccaccgcc gcagcggagg acaccccgcc gtggacgccc 5280
cgtggcaccg tcctcatcac cggcggcacc ggggccctgg gctcccgcgt cgcccaccgc 5340
atcgccgaac gccaccccga ctgccacttg ctgctggtga gccggcgagg gcccaaggcc 5400
cccggcgcca ccgcgctccg cgaccagctc atcgaactcg gcgccacggt gaccctcgcc 5460
gcatgtgaca ccgccgaccc cggcgcgctc gcggatctcc tcgccgatgt cccctcggac 5520
cgccccctca ccgcggtcgt ccacaccgcg ggcgtcctgg acgacagcac cctcgccgta 5580
cagaccccgg accacctcgc cgccgttctg gggcccaagt cccatgccgc acaccatctg 5640
cacgccctcg cccagcacca ccccctcgac gcgttcgtcc tcttctcgtc cgtcgcggcg 5700
cccttcggcg ccgcgggcca ggccaactac gcggccgcca acgcctacct cgacgccctc 5760
gcccagcacc gccgggccca ggggctggcc gccacctcca tcgcctgggg caactgggac 5820
ggcgacgggc tcgcgagcac acagtccgcg cagacgtacc tgcgcaaccg cggctttcct 5880
ccgatgccgc cacacctggc gctggccgcc ctggagcgag ccatcgtctc gccccacgcc 5940
cagctcgtcg tcgccgacgt cgactggaag aagctcaagc cggcgccgca cacccgcgac 6000
atcccgggaa gccgccgccc ggccccggcc gccaccgacg gcgcggacag gacggccgac 6060
gccaccgcga gcctccgtac ccgcctcgcg ggtcagagcc cggccgaacg gcaccagacg 6120
ctcctcgacc tcatcagctc tcatacagcc gccgtcctcg ggcacgccac gccccagacg 6180
atccccacgg accgggcctt ccgcgacctg ggtttcacct cgctgacggc catcgagctc 6240
cgcaaccgcc tcgcggcggc caccgggctc cgcctgccga ccaccgtcgc cttcgaccgt 6300
ccgacgccgg acaagctcgc ggccgacctg ctggcgcggt gcgcgccgac aggcccggac 6360
ggcatcgggg tgacgcccga cgcgacggcc acgagtggca gttcgcccgg tgcggcgcat 6420
ggcgcgccgg accccgccga gcccatcgcc atcgtcggct tggcctgccg ctaccccggc 6480
gggatcggct cccccgagga cctgtgggag ttcatcaccg cacaccggga cgccgtcgga 6540
gacttcccga ccgaccgggg ctgggacctg gcgaggctct tcgaccccga tccggaccgg 6600
ccgggcacct cgtacagccg acagggcgcc ttcctccgcg acgcgggcga cttcgacccg 6660
gagttcttcg ggatcagccc acgggaggcg acggcgacgg acccccagca gcgactgctc 6720
ctggaggcgt cctgggaagc cctcgaacga gccgggatca acccccacga tctccacggc 6780
agtccgacgg gcgtcttcac cggcagcaac gcgcaggact tcagcgcgcg gctgcggcag 6840
acgccgtcgg agctggcgga gctgtgcgag ggctatgcgc tgactggcag caacaacagc 6900
gtcgcctcgg ggcgcgtctc gtacgcgctc ggcctggaag gcccggcggt cagcatcgac 6960
accgcctgct cgtcctcgct cgtggcgctc catctggcct gccagtcgct ccgggccggc 7020
gaatgctcgc tggccctggc gggcggcgtc acggtcatga tgaccccgtt caacttcgtg 7080
gagttctccc ggcagcgggg cctggcggcg gacggccggt gcaaggcgtt ctccgccaca 7140
gccgatggca ccggctgggg cgagggcgtg ggcatggtgg tggtggagcg gctgtcggac 7200
gcgcggcgca acggccatcg tgtgctggcc ctcgtccgcg gcagcgccgt caaccaggac 7260
ggtgccagca atgggctgac tgccccgaac ggcccctcgc agcagcgggt catccgcgcc 7320
gccctggccg ccgccggggt cgccgcggca gaagtggacg cggtcgaggc gcacggcacg 7380
gggacgacgc tcggcgatcc gatcgaggcc caggccctgc tcgccaccta cgggcagggg 7440
cggccggcgg accgggcgct gtggctcggt acggtcaagt ccaacatcgg acacgcccag 7500
tcggccgccg gtatcgccgg ggtcatcaag atggtgctgg ccctgcggca cgggatgctg 7560
ccgcgtacgc tgcatgtgtc cgagccgtcg ccgcatgtgg actggtcggc gggtgcggta 7620
cggctgctga ccgaggacca gccgtggccg gacaccgggc gcccccggcg ggcgggggtg 7680
tcgtccttcg gcgtgagcgg caccaatgcc catgtgatcc tggagcaggc ggagccgggg 7740
ccggacccgg caccgacggc ctccgcgccc tccctgcccc cctggcccct ctccgccagg 7800
tcggcggagg ccctgcgggc ccaggcccgt aggttgctgg cgtacgtggc cgagcacccg 7860
gatgtcgacc ccgccgacgt ggggtactcc ctcgcgcgcg gacgggccgt gttcgagcac 7920
cgggccgtgc tcctcggcac cggccacgac gacttccggc gcgccctgga cgccctggcg 7980
tcgggcgcgc ccgacggcgc ggtcgtccag ggcgcggcgg tggggcggca gggcaaggtc 8040
gtctttgtgt gctcggggca gggcacccag cgccccggca tgggccgcgg gctctaccgc 8100
tcgtccacgg cgttcgccgg ggcgctggag gaggtgtgcg cccatctgga cccgtatctg 8160
gaacaccctc tgatggaggt gatgttcgcc gatgagaaga gcgatacgtc ggcgctgctg 8220
catctgaccg cctacgccca accggccctc ttcgccctcc agaccgcgct gcaccgcatg 8280
gtcaccgagg agttcgggct cacccccgac tatctggccg gccactccct gggcgagctg 8340
accgccgccc atctggcggg catcctcagc ctgcccgacg ccgcggcgct ggtggcggcc 8400
cgcgcccgcg ccatgcggga ccttccagcg accggagcca tggtcgccgt cgaggccacc 8460
gaggcggagc tgcggccccg gctcgccgag ttggcggacc gggtcggcat cgccgccgtc 8520
aacgcccccg cgtccctggt catcaccggc gaccacgacg ccgtgcacca gatcgccgac 8580
gacttccgcg ggcagggcag gaaggtcact cccctccagg tcagcggcgc cttccactct 8640
ccccatatgg agcccctgct cgacgagatc gggcgcaccg ccgaaaccct cacctaccac 8700
cggccccaca ctcccctcgt caccgcgtcg gccgacggcg gcgacgacac gaccgagccg 8760
cgggccgacg acgacccggg cacggccgcg ttctggcctc tccaggcccg gcgcaccgtc 8820
cactacgcgc gggccgtgga gcggctgcgc gcccgcggcg tcaccacgtt cctggaactc 8880
ggccccgact ccaccctcac taccctcgtc caccacaatc tcgccgcgca cgatcccgtg 8940
gccgtctccc tgctccatcc ggagcggtgc gagacgcaca gcgtcctcgg cgcactcgcc 9000
gcggtccacg cccacagccg ccccgtcgac tggacacgcc actacaccgc acggccgcgg 9060
ccgacgccac accagatcga cgtgcccacc tatgccttcc ggcaccggcg ctactggctg 9120
cccgccccgg cggcggtcgg cgatgtgacg gccgcggggc tcgacgcggc ggagcacccg 9180
ctgatcggcg ccgccgtgtg gctcgcggag ggcgacggct gtctgctgac cggcaggatc 9240
tcgccgcgta cgcacccgtg gctggccgac catgtcatcg ccggcactgt gctccttccg 9300
ggcaccgcgt tcgtggagct ggcgctgcgg gccggggcgt acgtgggctg cgaccgtgtg 9360
gaggagctga ccctgcacgc gccactcccg ctgcccgccg acggtgaggt ggtgctgcag 9420
gtggcggtgg gggccgccga cgagtccggc cgccgtgagc tgagcatcca cgcccggccg 9480
gcggacgacg gtacatggac acggcacgcc atcggcacgc tggcatcggc ccgcggcgtc 9540
ggcctcgacg atggcacggg gcacaatggc cacgccccgg cgggcgacga gccgttcggg 9600
tcgtgggcca cggcctggcc gccgcccggt gccgagccct tggacgtcac cggggtctac 9660
gaccggtttg ccgacgccga gttcacgtac ggggaggcat tccaggggct ggtcgcggct 9720
tggcggcacg gcgacgagac gctggcggag gtccgcctcc ccgaccagcc ggccggtgac 9780
gccctccgct tcgggctgca ccccgcgctg ctcgacgcgg cactgcagac catgtggctc 9840
gtggagcccg acggcacacg gccgagcggt ggcctgggcg gccccgatcg gggcctgccg 9900
ttcgcctggc agggggtctc gctgcgtacg gcgggcccgt cggccctgcg ggtacggctg 9960
cgacggccgg cgccggacac cgtggccgtc gccgtggccg acgcggccgg ccggccggtc 10020
gcgtcggtgg agtcgctgac gctgcggccg gtgccgcggg gcgccttgcg cggcaccgag 10080
acggcggtgc gcacctcgtt gtacggcctg gactggacgg atgtgccgct gccgacgccg 10140
cagacggccc tgccccggtg tgcgctgatc ggagcggaca cgctcgacct ggtccccgcg 10200
ctcgaggccg cggcgcccga ccgcatcacc gacggcgtgg agcgctacgc cgacctggag 10260
gagctggtgc gctccgtggc ggcgggcgcc cccgccccgg acctcgtcat cgccggctgc 10320
cacgcagccc ctgaagccga cggcgcgagc gaacagccac agcccgagac ggtgcgcaca 10380
aggacgggtc aggtgctgga gctgcttcag cggtggctcg gcgcggacgg gctcgccgac 10440
gcacacctgg tgctgttcac ctcaggcgcg gtcgccaccc ggccgggcga gccggtgcgg 10500
gacctggcgg gggcggcggt ctggggtctg gtgcgctccg gccagtcgga gcatccggag 10560
tgcttcaccg tggtggacat ggacggcgcc caggagtccc gcgcggcgct gctcggcgcg 10620
ctcggcctcg gcgagccgca actggcggtg cgcggcggcc gggcgctggc gccgcgcctg 10680
gtgcgcccgg gtgacgccga cgacgacagc ggcctggccc tgccgcaggg gccggaaggc 10740
tggcggttgg agtgtcccgg cacgggcagc ctggacgggt tgaccacgac cgagtccccg 10800
gccgcggcgg tgccgctcgg cccgggcgag gtacgggtcg cggtgcgggc cgcggggctg 10860
aacttccgcg atgtgctgat cgcgctgggc gtggtgcccg ggcggacggc gctgggcagt 10920
gagggggcgg ggatcgtcct cgaggtcggg gcggaggtcc gcgatctcgc gcccggggac 10980
cgggtggtgg gtatcttccc cgaggcgttc ggcccggtgg ccgtggccga gcgggcgacc 11040
ctggcgcggg tccccgacgg ctggtcgttc gcccaggccg cgtcggtccc catcgtgttc 11100
gccaccgcgt accacggcct ggtcgatctg gcgcgcctgc ggccggggga atcggtgctg 11160
atccatgccg cggccggcgg ggtgggcatg gccgccgtgc aactggcgcg ccatctgggg 11220
gccgaggtgt acgccacggc cggccccggc aagtggcaca tcctgcgttc ccaaggcatc 11280
gacgacgacc atctggcctc gtcgcgcacg ctggagttcg agcagcgctt cgccgcgacc 11340
cgcggcgggc gggggatcga tgtcgtcctg gactgtctgg cccatgagtt cgtcgacgcc 11400
tcgctgcgcc tggtggcgcg tgacggcggc cggttcctgg agatgggcaa gagcgacatc 11460
cgtgacccgc ggcaggtggc gctggaccat ccgggcgtgc tctaccgggc gttcgacctg 11520
ctggaggccg ggccggagcg ggtcgggcag atcctgcgca ccgtactgga cctgttcgag 11580
cgcggtgtcc tggcgcacct gccgacgacc tgctgggaca tccggcaggc ggagcacgcc 11640
ttccgccatc tgcagcaggg ccgtcacatc ggaaagaacg tgctcaccgt cccggccggc 11700
tggaacgccg agggcaccgt actgatcacc ggcggtatgg gcaccctggg cgccgccctc 11760
gcccgtcatc tggcgggtac cgggcgcgcc cgccatctgc tgctggccgg ccgacgcggc 11820
cccgacgccc cgggcgccga ggagctgcga gaggagctga ccgagctggg cgcgcgggtc 11880
accatcgccg catgcgatct cggcgaccgg gcggcggtcg cccggctcct gggggcgatc 11940
ccggccgagc ggccgctgac cgctgtcatc cacgcggcgg gtgtcgtcga cgatgccacc 12000
ctcgggtccc tcaccccccg ccacctggac gccgccctgg ccgccaaggc cgacgccgcc 12060
tggcatctgc acaccctcac ccgccacgcc gacgtggccg cgttcgtcct cttctcctcg 12120
gtcgcgggtc tgctcggctc gcccgggcag ggcaactacg ccgcggccaa cgccttcttg 12180
gacgcgctcg cccaccaccg gcgcggctct ggccttccgg cggtgtcgct ggcgtggggg 12240
ctgtgggagc agaccagcgg catgaccggg cacctggacc aggccgaccg cgcccggctg 12300
gcccggctcg gcatcagccc gctcacgacc gggcaggcgc tcggcctttt cgacgccgcc 12360
ctcggccacc accgccccgt gctcgtcccc gcccgcctcg acgtgcccga tccgcacccc 12420
ggctcgtcga ccgtgccgcc cctgtaccgg ggcctggtcg gatccaggac ccggcggaca 12480
ccccccgcgg ccgccgccac cgggccgttc cccctgcata cccgcctcgg cggtcacgcc 12540
ccggccgagc agcacgagat gctgctctcg ctggtccgct cccacgccgc cctcgtgctg 12600
ggccgcgacg atccggacac ggtccatccc ggcgcgcact tccgcggcct gggcttcgac 12660
tccctgaccg cggtcgagct ccgcaaccgg ctcaacgccg ccaccggcct ccggctctcc 12720
accaccctcg tcttcgacca ccccacgccc gacgaactcg cccgtcacgt ccgggagcag 12780
gtgctgggcg acggcgaagc ggcgcgggtg gccccggtgc tggccgagct cgacaggctg 12840
gaagcggcgc tgtcccgggt ggacggggac gatgcggtcc gggcgagggt gacggcccgg 12900
ttgcaggccc ttctcctgaa gtggaacgag tccgatggtc cggcgacggg cggtgacggt 12960
gcgggcaggc tggcgtccgc cacggccgcc gaggtgctgg atttcatcag gaacgacctc 13020
ggcctctcct ga 13032
<210> 69
<211> 12993
<212> DNA
<213> Artificial Sequence
<220>
<223> meilingmycin biosynthetic gene cluster of Streptomyces
nanchangensis strain NS3226
<400> 69
gtggccggac atccgtggat cctctccgga cacaccggaa ccgcgctgcg ggcccaggcg 60
cgccggctcc acgaccatgt cgccgaccac cccctgctcc gtccggaaga catcgcgcac 120
acgctggcga gcggcggccc ggcgctcacc catcgcgcgg cggtgatcgc ggcggaccgg 180
gagggatatc tccgggggct cgacgcggtg gcccgaggtg aggacgcccc cggtgtcgta 240
cggggcacgg cgaccgcggt cggcgacggg gtcgcgttcg tcttccccgg ccagggcacc 300
cagtggcccg gtatggccgc ggatctgctg acggtctccc ctgccttcag ccgggcggtc 360
gacgcctgcg ccgaggcgtt cgaaccgtat gtcccctggt caccggaggc ggtgctgcgg 420
ggcgctccgg gcgcgccgcc cctggagggg accgatgtgg tgcagccgac gctgttcgcc 480
gtcatggtgg ggctggccga gctgtggcgg actcttgggg tgagcccgac gacgatcgtg 540
gggcactgca tcggggagat cgcggcggcc catctctgcg gcgccctgtc gctgtccgac 600
gcggcgcgcg tggtgatcga gagcagccgg gcccaggcga cgctctccgg gtcgggtgcg 660
ctgatcgcgg tcgcgcggtc cgaggcgcag ctgcttccgc tactgcggcg gtggccgggc 720
aggctgacga tcgccgcggt caacggcccg atggccacgg tcgtctccgg cgatcggccg 780
gccgccgacg agctgttggc ggagttggcc cgtgccggtg tccgggcccg cgaggtggcg 840
atcgacatcc ccgcgcactc ggcgttcatg gcccccctca gggacggtct gctcgactcg 900
ctgtcatcgg tcaccgcggg tgcgtcgcgg ctgccgttcc actcctcggt catcgggggg 960
ccgctggaga cccaagggct cgacgcggct tactggtacc ggaacctcgc cgacacggtc 1020
cgcttcgaaa gcgtggtcac ggggctgctg cggcagggca cgcgctgctt cgtggagctg 1080
agcccgcatc cgatgctgac catgtgtgtg caggccaccg ccgaggaggt ggtcggcggt 1140
gagcgcgtcg tgatcctgcc gacgctgcat cgcgggcaag ccgccgtcga gtccgttcgc 1200
accacgctgg ccgagctgta cgtacggggg gcgctggatg accctcgggc ggcgttctcg 1260
gtgccgggcg gccgactgat caccctgccc ctcgagccgc tcgcggacac gtccgtagag 1320
ctcgccgacg ccccggatcc tgcggaggcc tgccggcccc cttgggcgga gcggcttgcc 1380
cggctctcca ccgcggagcg gaagcggcgg ctgtgcgagc tggtgggcgt cgaggcggcc 1440
aaggtcctcg aggacgtcgc cggggcggac gcgccgcgcc acggcatcgc tgagcaggag 1500
cacttcgtcg cttcgggctt cgactccgcg gccgcggtcg cgctgcgcaa ccgcctgaac 1560
gacgccaccg gactgctgct gcccttcacc ctggccttcg accatccgac acccgccgcc 1620
gtcgccgacc atctgcactc ccggctcttc gatcaccggg gcggtgggca gccgggcgcc 1680
gacggctggc ccgaccccgc ggcggcggcc ggtccggcca gggccgacga cgagccgatc 1740
gccgtcatcg gcatggcggg ccgcttcccc gggggcgctc gtaccccgga ggagctgtgg 1800
gatctggtcg ccgaaggcac cgacgccctc tcccccttcc cggagggccg gggctgggat 1860
ccgctgcggc tctacgatcc ggaccccgcc cggcccggca cgtactacca gcgcgaagcg 1920
ggattcctcc acgacgccga caagttcgac gccgagttct tcggcatcgc gccacgcgag 1980
gccacggcca tggatcccca gcagcggctg ctcctggaga cctcctggga ggcgctcgaa 2040
cgggcgcgga tcgacccgac cgcgctgcgc ggcagccgca ccggggtgtt cgtcggcgtg 2100
gccccgctgg actacagccc ccgtatgcac caggcgtcgc cggagctgga gggccatctg 2160
ctgaccggca acatcggcgc cgcggcctcg gggcggatct cctacgtact cgggctcgag 2220
gggcccgcgg tgtccgtgga cacggcgtgc tcgtcgtccc tggtagccct gcatctggcg 2280
gctcaggcgc tgcgggccgg ggagtgctcg ctggccctgg tcggcggggc gacggtcctc 2340
tcgacccccg gcatgttcat cgagttctcg cggcagcgcg gtctggctcc ggacggccgc 2400
tgcaaggcgt acgcggccgc cgcggacggc accggctggt ccgagggcgt gggcatgctg 2460
ctcgtcgagc ggctgtccga cgcacgacgg ctcggacacc aggtgctggc ggtggtacgg 2520
ggctccgccg tcaaccagga cggggcgagc aacggcttca cggcgcccag cggtccatca 2580
cagcaacagg tcatccgggc ggccctggcc aatgcggggg tgtcggctcc ggaggtcgac 2640
gcggtggagg ggcacggcac cggcacccgg ttgggcgatc cgatcgaggc gcaggcgctg 2700
ctggcggcgt acgggcaggg gcgggcggcc gaccggccgc tgtggctggg ctcgatcaag 2760
tcgaacatcg gacacaccca gtgggccgcg ggcgtcatcg gggtcatcaa aatggtgctc 2820
gcgctccagc gcggtgtgct gccgcgcacg ctgcacgtgg acaagccgtc ggattacgtg 2880
gactggtcgg cgggggccgt acggctgttg acggagccgg tgccctggcc ggagaggggc 2940
cacccgcgcc gggcgggggt gtcgtccttc ggcgtgagcg gcaccaacgc ccatgtcatc 3000
ctcgagcagg caacgccatc gtccacggtg gctcccgagg ggcctaccgc cgaggccggg 3060
cctcccctgc cgtgggtgat ctcggcgaag accccccagg cactgcgcga ccaggcccgc 3120
cgcctgcacg aacacctcac cgcccagcca cagctccaac cggccgacgt cggccacacc 3180
ctcgccaccg gccgcgccac cttcgaccac cgggccgtcc tcatcggctc cgaccgcgaa 3240
caactcctcc acggcctgga cgcgctcgcc accggccggc ccgacccagc ggtccaccag 3300
acagcggacc gtcccgccac cgccgacggc cgtatcgtct tcgtcttccc cggacaaggc 3360
ggtcaatggg cgggcatggg tctacggctg ctgaacgcct cacccgtctt caccgagcgg 3420
atggccgcct gcgaacaggc cctctccccc tacgtcgact ggtcactcac ggacatcctc 3480
caccggccgg ccgacgacgc cgcatggcaa cgcgccgaca tcgtccagcc cgccctgttc 3540
tcgatcatgg tgtccctggc cgcgctctgg cgctcttgcg gcatcgaacc ggacgcggtc 3600
ctcggccact cccaaggcga gatcgccgcg gcccacgtct gcggcgcact gacgctccac 3660
gacgcggcca aggtcatcgc cctgcgcagc caggccctcc aagccgtacg cggcgccggg 3720
ggcatggcct ccgtacccct gtccgcggac caggtcaccg aggatctgca cacccactgg 3780
cccgaccggc tctgggtggc cgccaccaac tcccccacgg caaccgtcat ctcgggaaac 3840
accgacgcac tcgacgaagc gctcgaccac taccacgccc acgacgtacg ggccaaacgc 3900
atcccggtcg actacgcctc ccactgcccc catatcgacg cggtggccga gcgactgccc 3960
gatctgctgg gcggcatcgt cccgcgcgcc gccgacatcc ccttctactc cacggttgac 4020
ggccgatggg ccgagccgac cgagctcgac gccgactact ggtaccgcaa cctccgcagc 4080
cccgtacggt tcgcccacgc cgtccacgcc ctcaccgagg ccgaacaccg caccttcgtc 4140
gaagtcagcc cacaccccac gctcaccccc gccatcacgg ccaccgccga aaccaccgac 4200
cgcaccacca ccgtcatcgc ctcgctccac cgcgaccacg aagacgctca ccacatcctc 4260
accaacctcg cccaggccca catccacggc cacaccgtcg cctggcgaca ccactaccgg 4320
actctgcgcc ccaccccgcc ccacatcgac ctccccacct accccttcca acaccagcac 4380
tactggctcc acgactccac cgaggacaag gcggtgggta cggacctcgc tgcggcccgc 4440
ttctgggagg cagtcgacgg cgaggacacc aacgccgtcg ccgcgctcct cgacgtcgag 4500
ccgggcacct cgctggacgc gctgctgccg gccctgtccg cctggcacgg tcggcgtcgc 4560
gaccaggcca tcaccgacac ctggtgttac cgggacatct ggaagccggt cgacctcacc 4620
gccgcgcgcc cccgaccgtc cagccgatgg cttgtcgcga tctccgcagg gcgggccgat 4680
cacctccacg tcagtgccgt cctggacgct ctggaacgcc agggtctgcc catcgccacc 4740
ctcgtcctcg acgacaccca catcgaactc cccctgctgg agcggcatct cgcacaggtg 4800
atcgcgagcg atgggccggc catcggcggc gtgctctcgc tgctcgccct cgacgagggg 4860
ccacatccgc gccacccgga ggtgcccgtc ggcaccgccc tcaccctcag cctgatccag 4920
gcgctcatcg cacgtgagga catcgcgccc cggctctggc tggccaccca cgaggccgtc 4980
gccacctcgt ccgcggatac gctcgatcac cccctccagg cgatggtctg ggggctggga 5040
cgcaccgccg ccctcgaaca ccccgatctg tggggcggac tcatcgacct tccggacact 5100
ctcaccgaac gggtcctccg cggcctcgtc acggcgctga ccacctgtca cgacgaggac 5160
gagctcgcgc tgcgcgccac cggcccacgc acccggcggc tggtccggac gccgtccacc 5220
gccgcggcgg aggacacccc gccgtggacg ccccgtggca ccgtcctcat caccggcggc 5280
accggggccc tcggctcccg cgtcgcccac cgcatcgccg aacgtcaccc cggctgccac 5340
ttgctgctgg tgagccggcg aggggccaac gcccccggcg ccaccgcgct ccgcgaccag 5400
ctcatcgaac tcggcgccac ggtgaccctc gccgtatgtg acaccgccga ccccggcgcg 5460
ctcgcggatc tcctcgccga tgtcccctcg ggccgccctc tcaccgcggt cgtccacacc 5520
gcgggcgtcc tggacgacag caccctcgcc gtacagaccc cggaccacct cgccgccgtt 5580
ctggggccca agtcccatgc cgcacaccat ctgcacgccc tcgcccagca ccaccccctc 5640
gacgcgttcg tcctcttctc gtccgtcgcg gcgcccttcg gtgccgcggg ccaggccaac 5700
tacgcggccg ccaacgccta cctcgacgcc ctcgcccggc accgccgggc ccaggggctg 5760
gccgccacct ccatcgcctg gggcaactgg gacggcgacg ggctcgcgag cacccagtcc 5820
gcgcagacgt acctgcgcaa ccgcggcttt cctcccatgc cgccacacct ggcgctggcc 5880
gccatggagc gagcggtcgt ctcgccccac gcccagctcg tcgtcgccga cgtcgactgg 5940
aagaagctca agccgacgcc gcacacccgc gacatcccgg aaagccgccg cccggccccg 6000
gccgccaccg acggcgcaga caggaccgcc gacgccaccg cgagcctccg tacccgcctc 6060
gcgggtcaga gcccggccga acggcaccag acgctcctcg acctcatcag ctctcataca 6120
gccgccgtcc tcgggcacgc cacgccccag acgatcccca cggaccgggc cttccgcgac 6180
ctgggtttca cctcgctgac ggccatcgag ctccgcaacc gcctcgcggc ggccaccggg 6240
ctccgcctgc cgaccaccgt cgccttcgac cgcccgacgc cggacaagct cgcggcggac 6300
ctgctggcgc ggtgcgcgcc gacgggcccg gacggcatcg gagtgacagc cgacgcgacg 6360
gccgcgagcg gcagttcgcc cggtccggcg catggcgcgc tggaccccgc cgagcccatc 6420
gccatcgtcg gctgggcctg ccgctacccc ggcgggatcg gctcccccga ggacctgtgg 6480
gagttcgtca ccgcacaccg ggacgccgtc ggagacttcc cgaccgaccg gggctgggac 6540
ctggcgaggc tcttcgaccc cgatccggac cggccgggca cctcgtacag ccgacagggc 6600
gccttcctcc acgacgcggg cgacttcgac ccggagttct tcgggatcag cccacgggag 6660
gcgacggcga cggaccccca gcagcggctg ctcctggaga cgtcctggga agccctcgaa 6720
cgagccggga tcaacccgca cgatctccac ggcagtccga cgggcgtctt caccggcagc 6780
aacgcgcagg acttcagcgc acggctgcgg cagacgccgt cggagctggc ggagctgtgc 6840
gagggctatg cgctgacggg cagcaacaac agcgtcgcct cggggcgcgt ctcgtacgcg 6900
ctcggcctgg aaggcccggc ggtcagcatc gacaccgcct gctcgtcctc gctcgtggcg 6960
ctccatctgg cctgccagtc gctccgggcc ggcgaatgct cgcttgccct ggcgggcggc 7020
gtcacggtca tgatgacccc gttcaacttc gtggagttct cccggcagcg gggcctggcg 7080
gcggacggcc ggtgcaaggc gttctccgcc accgccgatg gcaccggctg gggcgagggc 7140
gtgggcatgg tggtggtgga gcggctgtcg gacgcgcggc gcaacggcca tcgtgtgctg 7200
gccctggtcc gcggcagcgc cgtcaaccag gacggtgcca gcaatgggct gactgccccg 7260
aacggcccct cgcagcagcg ggtcatccgc gccgccctgg ccgccgccgg ggtcaccgcg 7320
gcagaggtgg acgcggtcga ggcgcacggc acggggacga cgctcggcga tccgatcgag 7380
gcccaggccc tgctcgccac ctatgggcag gggcggccgg cggaccgggc gctgtggctc 7440
ggtacggtca agtccaacat cggacacgcc cagtcggccg ccggtatcgc cggggtcatc 7500
aagatggtgc tggccctgcg gcacgggatg ctgccgcgta cgctgcatgt gtccgagccg 7560
tcgccgcatg tggactggtc ggcgggtgcg gtacggctgc tgaccgagga ccagccgtgg 7620
ccggacaccg ggcgcccccg gcgggcgggg gtgtcgtcct tcggcgtgag cggcaccaac 7680
gcccatgtga tcctggagca ggcggagccg gggccggacc cggacccggc gccgacggcc 7740
tccgcgcact ccgtgctccc ctggcccctc tccgccaggt cggcggaggc cctgcgggcc 7800
caggcccgta ggttgcgggc gtacgtggcc gagcacccgg atgtcgaccc cgccgacgtg 7860
gggtactccc tcgcgcgcgg acgggccacc ttcgagcacc gggccgtgct cctcggcacc 7920
ggccacgacg acttccggcg cggcttggac gccctggtgt cgggcgcgcc cgacggcgcg 7980
gtcgtccagg gcgcggcggt ggggcggcag ggcaaggtcg tctttgtgtg ctcggggcag 8040
ggcacccagc gccccggcat gggccgcggg ctctaccgct cgtccacggc gttcgccggg 8100
gcgctggagg aggtgtgcgc ccatctggac ccgtatctgg aacaccctct gatggaggtg 8160
atgttcgccg acgagaagag cgatacgtcg gcgctgctgc atctgaccgc ctacgcccaa 8220
ccggccctct tcgccctcca gaccgcgctg catcgcatgg tcaccgagga gttcgggctc 8280
acccccgact atctggccgg ccactccctg ggcgagctga ccgccgccca tctggcgggc 8340
atcctcagcc tgcccgacgc cgcggcgctg gttgcggccc gcgcccgcgc catgcgggac 8400
cttccggcgg ccggagccat ggtcgccgtc gaggccaccg aggccgaact gcggcctcgg 8460
ctcgccgagt tggcggagcg ggtcgacatc gccgccgtca acgcccccgc gtccctggtc 8520
atcaccggcg accacggcgc cgtgcaccag atcgccgacg acttccgcgc gcagggcagg 8580
aaggtcacct ccctccaggt cagcggcgcc ttccactccc cccatatgga gcccctgctc 8640
gacgagatcg ggcgcaccgc cgaaaccctc acctaccacc ggccccacac tctcctcgtc 8700
accgcatcgg cggacggcgg cgacgacacg atcgagccgc gggccgacga cgacccgggc 8760
acggccgcgt tctggcctct ccaggcccgg cgcaccgtgc actacgcacg ggccgtggag 8820
cggctgcacg cccgcggcgt caccacgttc ctggaactcg gccccgacgc caccctcacc 8880
gccctcgtcc accacaacct cgccgcgcac gatcccgtgg ctgtctccct gctccatccg 8940
gagcggtgcg agacgcacag cgtcctcggc gcgctcgccg cggtccacgc ccacagccgc 9000
cccgtcgact ggacgcgcca ctacaccgca cggccgcggc cgacgccaca ccagatcgac 9060
gtgcccacct atgccttccg gcaccggcgc tactggctgc ccgccccggc ggcggtcggc 9120
gatgtgacgg ccgcggggct cgacgcggcg gagcacccgc tgatcggcgc cgccgtgggg 9180
ctcgcggagg gcgacggctg tctgctgacc ggcaggatct cgccgcgtac gcacccgtgg 9240
ctggccgacc atgtcatcgt cggcaccgtg ctgcttccgg gcaccgcgtt cgtggagctg 9300
gcgctgcggg ccggggcgta tgtgggctgc ggccgtgtgg aggagctgac cctgcacgcg 9360
ccgctccccg ccgacggtga ggtggtgctc caggtgacgg tgggggccgc cgacgagtcc 9420
ggccgccgtg agctgagcat tcacgcccgg ccggcggacg acggtacatg gacacggcac 9480
gccatcggca cgctggcacc ggcccacgac gtcgacgcgg gtcaagatgg ccacgccccg 9540
gcggatgacg ggcagttcgg gtcgtgggcc acggcctggc cgccgcccgg tgcggagccc 9600
ttggacgtca ccggggtcta cgcccggttt gccgacgccg agttcacgta cggggaggcc 9660
ttccaggggc tggtcgcggc ttggcggcac ggcgacgaga cgctggcgga ggtccgcctc 9720
cccgaccagc cggccggtga cgcccaccgc ttcgggctgc accccgcgct gctcgacgcg 9780
gcactgcaga ccatgtggct cgtggagccc gacggcacac ggccgacggg tggcctgggc 9840
ggccccgatc ggggcctgcc gttcgcctgg cagggggtct cgctgcgtac ggcgggcccg 9900
tcggccctgc gggtacggct gcgacggccg gcgccggaca ccgtggccgt cgccgtggct 9960
gacccggccg gccgaccggt cgcgtcggtg gagtcgctga cgctgcggcc ggtgccgcgg 10020
ggcgccttgc gcggcgccga ggcggcggtg cgcacctcgt tgcacggcct ggactggacg 10080
gatgtgccgc tgccgacgcc gcccccggcc cggccccggt gtgcgctgat cggagcggac 10140
acgctcggcc tgggccccgc gctcgaggcc gcggcgcccg accgcatcac cgacggcgtg 10200
gagcgctacg ccgacctgga ggagctggtg cgctccgtgg cggcgggcgc ccccgccccg 10260
gacctcgtca tcgccacctg ccacacagcc cctgaagccg acggcgcgag cgaacagcca 10320
cagcccgaga cggtgcgcac aaggacgggt caggtgctgg agctgcttca gcggtggctc 10380
ggcgcggacg ggctcgccga cgcacacctg gtgctgttca cctcaggcgc ggtcgccacc 10440
cggccgggcg agctggtgcg tgacctggcg ggggcggccg tctggggtct ggtgcgctcc 10500
ggccagtcgg agcatccgga gtgcttcacc gtggtggaca tggacggcgc ccaggagtcc 10560
cgcgcggcgc tgctcggcgc gctcggcctc ggcgagcctc aactggcggt gcgcggcggc 10620
cgggcgctgg cgccgcgcct ggtgcgcccg ggtgccgcag ccgacgacag cggcctggcc 10680
ctgccgcggg ggccggaagg ctggcggttg gagtgtcccg gcacgggcag cctggacggg 10740
ttgaccacga ccgagtcccc ggccgcggcg gtgccgctcg gcccgggcga ggtacgggtc 10800
gcggtgcggg ccgcggggct gaacttccgc gatgtgctga tcgcgctggg cgtggtgccc 10860
gggcggacgg cgctgggcag tgagggggcg gggatcgtcc tcgaggtcgg ggcggaggtc 10920
cgcgatctca cgcccgggga ccgggtggtg ggtatcttcc ccgaggcgtt cggcccggtg 10980
gccgtggccg agcgggcgac cttggcgcgg atccccgacg gctggtcgtt cgcccaggcc 11040
gcgtcggtcc ccatcgtgtt cgccaccgcg taccacggcc tggtcgatct ggcgcgcctg 11100
cggccggggg aatcggtgct gatccatgcc gcggccggcg gggtgggcat ggccgccgtg 11160
caactggcgc gccatctggg ggccgaggtg tacgccacag ccggccccgg caagtggcac 11220
atcctgcgct cccaaggcat cgacgacgac catctggcgt cgtcgcgcac gctggagttc 11280
gagcagcgct tcgccgcgac ccacggcggg cggggcatcg atgtcgtcct ggactgtctg 11340
gcccatgagt tcgtcgacgc ctcgctgcgc ctggtggcgc gtgacggcgg ccggttcctg 11400
gagatgggca agagcgacat ccgtgacccg cggcaggtgg cgctggacca tccgggcgtg 11460
ctctaccggg cgttcgacct gttggaggcc gggccggagc gggtcgggca gatcctgcgc 11520
accgtactgg acctgttcga gcgcggtgtc ctggcgcacc tgccgacgac ctgctgggac 11580
atccggcagg cggagcaggc cttccgccat ctgcagcagg gccgccacat cggaaagaac 11640
gtgctcaccg tcccggccgg ctggaacgcc gagggcaccg tactgatcac cggcggtacg 11700
ggcaccctgg gtgccgccct cgctcgccat ctggcgggta ccgggcgcgc ccgccatctg 11760
ctgctggtcg gccgacgcgg ccccgacgcc ccgggcgccg aggagctgcg agaggagctg 11820
accgagctgg gcgcgcgggt caccatcgcc gcatgcgatc tcggcgaccg ggcggcggtc 11880
gcccggctcc tgggggcgat cccggccgag cggccgctga ccgccgtcat ccacgcggcg 11940
ggtgtcgtcg acgatgccac cctcgggtcc ctcacccccc gccacctgga cgccgccctg 12000
gccgccaagg ccgacgccgc ctggcatctg cacaccctca cccgccacgc cgacgtggcc 12060
gcgttcgtcc tcttctcctc ggtcgccggt ctgctcggct cgcccgggca gggcaactac 12120
gccgcggcca acgccttctt ggacgcgctc gcccaccacc ggcgctgctc tggccttccg 12180
gcggtgtcgc tggcgtgggg gctgtgggag cagaccagcg gcatgaccgg agacctggac 12240
caggccgacc gcgcccggct ggcccggctc ggcatcagcc cgctcacgac cgggcaggcg 12300
ctcgaacttt tcgacaccgc cctcggccac caccgccccg tgctcgtccc cgcccgcctc 12360
gacgtgcccg acccgcaccc cggctcgtcg accgtgccgc ccctgtaccg gggcctggtc 12420
ggatccagga cccggcggac accccccgcg tccgccgcca ccgggccgtt ccccctgcat 12480
acccgcctcg acggtcacgc cccggccgag cagcacgaga tgctgctctc gctggtccgc 12540
tcgcacgccg ctctcgtgct gggccgcgac gatccggaca cggtccatcc cggcgcgcac 12600
ttccgcggtc tgggcttcga ctccctgacc gcggtcgagc tccgcaatcg gctcaacgcc 12660
gccaccggcc tccggctctc caccaccctc gtcttcgacc accccacgcc cgacgaactc 12720
gcccgtcacg tccgggagca ggtgctgggc gacggcgaag cggcgcgggt ggccccggtg 12780
ctggccgagc tcgacaggct ggaggccgcg ctgtcccggg tgaacgggga cgatgcgctc 12840
cgggcgaggg tgacggcccg gctgcaggcc cttctcctga agtggaacga gtccgatggt 12900
ccggcgacgg gcgcagacgg tgcgggcagg ctggcgtccg ccacggccgc cgaggtgctg 12960
gatttcatca ggaacgacct cggcctctcc tga 12993
<210> 70
<211> 13032
<212> DNA
<213> Artificial Sequence
<220>
<223> milA1 gene of Streptomyces milbemycinicus
<400> 70
ttgcccaaag cccagaacga gttcgcagtg gccggtcatc cgtggatcct ctccgggcac 60
accggaaccg cgctgcgggc ccaggcacgc cggctccacg accatgtcgc cgaccaccct 120
cggctccgtc cggaagacat cgcccacacg ctggcgagca gcggcccggc gctcacccat 180
cgcgcggcgg tgatcgcggc ggaccgggaa ggacatctcc gggggctcga cgcggtggcc 240
cggggtgagg acacccccgg tgtcgtacgg ggcacggcgg ccgcgggcgg cgacggggtc 300
gcgttcgtct tccccggcca gggcacccag tggcccggta tggccgccga tctgctgacg 360
gtctcccccg ccttcagccg ggcggtcgac gcctgcgccg aggcgttcga accgtatgtc 420
tcctggtcac cggaggccgt gctgcggggc gctccgggcg cgccgcccct ggaggggacc 480
gatgtggtgc agccgacgct gttcgccgtc atggtggggc tggccgagct gtggcggact 540
cttggggtga gcccgacgtc gatcgtgggc cactgcatcg gggagatcgc ggcagcccat 600
ctctgcggcg ccctgtcgct gtccgacgcg gcgcgcgtgg tgatcgagag cagccgggcc 660
caggcgacgc tctccgggtc gggtgcgctg atcgcggtcg cgcggtccga ggcgcagctg 720
cttccgttgc tgcggcggtg gccgggcagg ctgacgatcg ccgcggtcaa cggcccgatg 780
gccacggtcg tctccggcga tcggccggcc gccgacgagc tgttggcgga gttcgcccgt 840
gccggtgtcc gggcccgcga ggtggcgatc gacatccccg cgcactcgcc gttcatggcc 900
cccctcaggg acggtctgct cgactcgctg tcatcggtca ccgcgggtgc gtcgcggctg 960
ccgttccact cctcggtcat cggggggccg ctggagaccc aagggctcga cgcggcttac 1020
tggtaccgga acctcgccga cacggtccgc ttcgaaagcg tcgtcacggg gctgctgcgg 1080
cagggcacac gctgcttcgt ggagctgagc ccgcacccga tgctgaccat gtgtgtgcag 1140
gccaccgccg aggaggtggt cggcggtgag cgcgtcgtga tcctgccgac gctgcatcgc 1200
gggcaggccg ccgtcgagtc cgttcgcacc acgctggccg agctgtacgt acggggcgca 1260
ctggatgacc atcgggcggc gttctcggtg ccgggcggcc gcctgatcac cctgcctctc 1320
gagccgcccg cggacacgtc cgtagagctc gccgacgccc cggacccggc ggaggcctgc 1380
cggcccccct tggtggagcg gcttgcccgg ctctccaccg cggagcggaa gcggcggctg 1440
cgcgagctgg tgggcgtcga ggcggccaag gtcctcgagg acgtcgccgg ggcggacgcg 1500
ccgggccacg gcatcgcgga gcaggagcac ttcgtcactt cgggcttcga ctccgcggcc 1560
gcggtcgcgc tgcgcaaccg cctgaacgac gccaccggtt tgctgctgcc cttcaccctg 1620
gccttcgacc atccgacacc cgccgccgtc gccgaccatc tgcactcccg gctcttcgat 1680
caccagggcg gcgggcagcc gggcgccgac ggccggcccg accccgcggc ggcggccggt 1740
ccggccaggg ccgacgacga gccgatcgcc gtcatcggca tggcgggccg cttccccggg 1800
ggcgcccgta ccccggagga gctgtgggaa ctggtcgccg aaggcaccga cgccctctcg 1860
cccttcccgg agggccgggg ctgggatccg ctgcggctct acgatccgga ccccgcccgg 1920
cccggcacgt actaccagcg cgaagcggga ttcctccacg acgccgacaa gttcgacgcc 1980
gagttcttcg gcatcgcgcc acgcgaggcc accgcaatgg atccccagca gcggctgctc 2040
ctggagacct cctgggaggc gctcgaacgg gcgcggatcg acccgaccgc gctgcgcggc 2100
agccgcaccg gggtgttcgt cggcgtggcc ccgctggact acagcccccg aatgcaccag 2160
gcgtcgccgg agctggaggg ccatctgctg accggcaaca tcggcgccgc ggcctcgggg 2220
cggatctcct acgtactcgg gcttgagggg cccgcggtgt ccgtggacac ggcgtgctcg 2280
tcgtccctgg tcgccctgca tctggcggcc caggcgctgc gggccgggga gtgctcgctg 2340
gccctggtcg gcggggcgac ggtcctctcg acccccggca tgttcatcga gttctcgcgg 2400
cagcgcggtc tggctccgga cggccgctgc aaggcgtacg cggccgccgc ggacggcacc 2460
ggctggtccg agggtgtggg catgctgctc gtcgagcggc tgtccgacgc gcgacggctc 2520
ggacaccagg tgcttgcggt ggtacggggc tccgccgtca accaggacgg ggcgagcaac 2580
ggcttcacgg cgcccagcgg tccatcacag caacaggtca tccgggcggc cctggccaat 2640
gccggggtgt cggctccgga ggtcgacgcg gtggaggggc acggcaccgg cacccggttg 2700
ggcgatccga tcgaggcgca ggcgctgctg gccgcctacg ggcaggggcg ggcggccgac 2760
cggccgctgt ggctggggtc gatcaagtcg aacatcggac acacccagtg ggccgcgggt 2820
gtcatcgggg tcatcaaaat ggtgctcgcg ctccagcacg gtgtgctgcc gcgcacgctg 2880
cacgtggaca agccgtcgga ttacgtggac tggtcggccg gggccgtacg gctgttgacg 2940
gagccggtgc cctggccgga gcggggccac ccgcgccggg cgggggtgtc gtccttcggg 3000
gtgagcggca ccaacgccca tgtcatcctc gagcaggcaa cgccatcgtc cacggtggct 3060
cccggggggc ataccgccga ggccgggcct cccctgccgt gggtggtctc ggcgaagacg 3120
ccccaggcac tgcgcgacca ggcccgccgc ctgcacgaac acctcaccgc ccagccacag 3180
ctccaaccgg ccgacgtcgg ccacaccctc gccaccggcc gcgccacctt cgaccaccgg 3240
gccgtcctca tcggctccga ccgcgaacaa ctcctccacg gcctggacgc gctcgccacc 3300
ggccggcccg acccagcggt ccaccagacg tcggaccgtc ccgccaccgc cgacggccgt 3360
atcgtcttcg tcttccccgg acaaggcggt caatgggcgg gcatgggcct acggctgctg 3420
aacgcctcac ccgtcttcac cgagcggatg gccgcctgcg aacaggccct ctccccctac 3480
gtcgactggt cactcacgga catcctccac cggccggccg acgacgccgt atggcaacgc 3540
gccgacatcg tccagcccgc cctgttctcg atcatggtgt ccctggccgc gctctggcgc 3600
tcttgcggca tcgaaccgga cgccgtcctc ggccactccc aaggcgagat cgccgcggcc 3660
cacgtctgcg gcgccctgac gctccacgac gcggccaagg tcatcgccct gcgcagccag 3720
gccctccaag ccgtacgcgg cgccgggggc atggcctccg tacccctgcc cgcggaccag 3780
gtcaccgagg atctgcgcac ccactggccc gaccggctat gggtggccgc caccaactcc 3840
cccacggcaa ccgtcatctc gggaaacacc gacgcgcttg acgaagcgct cgaccactac 3900
cacgcccacg acgtacgggc caagcgcatc ccggtcgact acgcctccca ctgcccccat 3960
atcgacgcgg tggccgagcg actgcccgac ctgctgggcg gcatcgtccc gcgcgccgcc 4020
gacatcccct tctactccac ggttgacggc cgatgggccg agccgaccga gctcgacgcc 4080
gactactggt accgcaacct ccgcagcccg gtacggttcg cccacgccgt ccacgccctc 4140
accgagaccg accaccgcac ctttgtcgaa gtcagcccac accccacgct cacccccgcc 4200
atcacggcca ccaccgaaac caccgaccgc accaccaccg tcatcgcctc gctccaccgc 4260
gaccacgacg acacccacca catcctcacc aacctcgccc aggcccacat ccacggccac 4320
accatcgact ggcgacacca ctaccagact ctgcgcccca ccccacccca tatcgacctc 4380
cccacctacc ccttccaaca ccaccactac tggctccacg actccaccga ggacaaggcg 4440
gtgggtacgg acctcgccgc ggcccgcttc tgggaggcgg tccacggcga ggacaccaac 4500
gccgtcgccg cgctcctcga cgtcgagccg ggcacctcac tggacgcgct gctgccggcc 4560
ctgtccgcct ggcacggtcg gcgtcgcgac caggccatca ccgacacctg gtgttaccgc 4620
gacatctgga agccggccga cctcaccgcc gcgcgccccc ggccgtccgg ccgatggctt 4680
gtcgcgatct ccgcagggcg ggccgatcac ctccacgtca gtgccgtcct ggacgctctg 4740
gaacgccagg gtctgcccat cgccaccctc gtcctcgacg acacccacac cgaactcccc 4800
ctgctggagc ggcatctcgc acaggcgatc gcgagcgatg ggccggccat cggcggcgtg 4860
ctctcgctgc tcgccctcga cgaggggcca catccgcgcc acccggaggt gcccgtcggc 4920
accgccctca ccctcagcct gatccaggcg ctcatcgcac gcgaggacat ggcgccccgg 4980
ctgtggctgg ccacccacga ggccgtcgcc acctcgtccg cggatacgct cgatcacccc 5040
ctccaggcga tggtctgggg gctgggacgc accgccgcac tcgaacaccc cgatctgtgg 5100
ggcggcctca tcgaccttcc ggacactctc accgaacggg tcctccacgg cctcgtcacg 5160
gcgctgacca cctgtcacga cgaggacgaa ctcgcgctgc gcgccaccgg cccacgcacc 5220
cggcgcctga tccggacgcc gtccaccgcc gcagcggagg acaccccgcc gtggacgccc 5280
cgtggcaccg tcctcatcac cggcggcacc ggggccctgg gctcccgcgt cgcccaccgc 5340
atcgccgaac gccaccccga ctgccacttg ctgctggtga gccggcgagg gcccaaggcc 5400
cccggcgcca ccgcgctccg cgaccagctc atcgaactcg gcgccacggt gaccctcgcc 5460
gcatgtgaca ccgccgaccc cggcgcgctc gcggatctcc tcgccgatgt cccctcggac 5520
cgccccctca ccgcggtcgt ccacaccgcg ggcgtcctgg acgacagcac cctcgccgta 5580
cagaccccgg accacctcgc cgccgttctg gggcccaagt cccatgccgc acaccatctg 5640
cacgccctcg cccagcacca ccccctcgac gcgttcgtcc tcttctcgtc cgtcgcggcg 5700
cccttcggcg ccgcgggcca ggccaactac gcggccgcca acgcctacct cgacgccctc 5760
gcccagcacc gccgggccca ggggctggcc gccacctcca tcgcctgggg caactgggac 5820
ggcgacgggc tcgcgagcac acagtccgcg cagacgtacc tgcgcaaccg cggctttcct 5880
ccgatgccgc cacacctggc gctggccgcc ctggagcgag ccatcgtctc gccccacgcc 5940
cagctcgtcg tcgccgacgt cgactggaag aagctcaagc cggcgccgca cacccgcgac 6000
atcccgggaa gccgccgccc ggccccggcc gccaccgacg gcgcggacag gacggccgac 6060
gccaccgcga gcctccgtac ccgcctcgcg ggtcagagcc cggccgaacg gcaccagacg 6120
ctcctcgacc tcatcagctc tcatacagcc gccgtcctcg ggcacgccac gccccagacg 6180
atccccacgg accgggcctt ccgcgacctg ggtttcacct cgctgacggc catcgagctc 6240
cgcaaccgcc tcgcggcggc caccgggctc cgcctgccga ccaccgtcgc cttcgaccgt 6300
ccgacgccgg acaagctcgc ggccgacctg ctggcgcggt gcgcgccgac aggcccggac 6360
ggcatcgggg tgacgcccga cgcgacggcc acgagtggca gttcgcccgg tgcggcgcat 6420
ggcgcgccgg accccgccga gcccatcgcc atcgtcggct tggcctgccg ctaccccggc 6480
gggatcggct cccccgagga cctgtgggag ttcatcaccg cacaccggga cgccgtcgga 6540
gacttcccga ccgaccgggg ctgggacctg gcgaggctct tcgaccccga tccggaccgg 6600
ccgggcacct cgtacagccg acagggcgcc ttcctccgcg acgcgggcga cttcgacccg 6660
gagttcttcg ggatcagccc acgggaggcg acggcgacgg acccccagca gcgactgctc 6720
ctggaggcgt cctgggaagc cctcgaacga gccgggatca acccccacga tctccacggc 6780
agtccgacgg gcgtcttcac cggcagcaac gcgcaggact tcagcgcgcg gctgcggcag 6840
acgccgtcgg agctggcgga gctgtgcgag ggctatgcgc tgactggcag caacaacagc 6900
gtcgcctcgg ggcgcgtctc gtacgcgctc ggcctggaag gcccggcggt cagcatcgac 6960
accgcctgct cgtcctcgct cgtggcgctc catctggcct gccagtcgct ccgggccggc 7020
gaatgctcgc tggccctggc gggcggcgtc acggtcatga tgaccccgtt caacttcgtg 7080
gagttctccc ggcagcgggg cctggcggcg gacggccggt gcaaggcgtt ctccgccaca 7140
gccgatggca ccggctgggg cgagggcgtg ggcatggtgg tggtggagcg gctgtcggac 7200
gcgcggcgca acggccatcg tgtgctggcc ctcgtccgcg gcagcgccgt caaccaggac 7260
ggtgccagca atgggctgac tgccccgaac ggcccctcgc agcagcgggt catccgcgcc 7320
gccctggccg ccgccggggt cgccgcggca gaagtggacg cggtcgaggc gcacggcacg 7380
gggacgacgc tcggcgatcc gatcgaggcc caggccctgc tcgccaccta cgggcagggg 7440
cggccggcgg accgggcgct gtggctcggt acggtcaagt ccaacatcgg acacgcccag 7500
tcggccgccg gtatcgccgg ggtcatcaag atggtgctgg ccctgcggca cgggatgctg 7560
ccgcgtacgc tgcatgtgtc cgagccgtcg ccgcatgtgg actggtcggc gggtgcggta 7620
cggctgctga ccgaggacca gccgtggccg gacaccgggc gcccccggcg ggcgggggtg 7680
tcgtccttcg gcgtgagcgg caccaatgcc catgtgatcc tggagcaggc ggagccgggg 7740
ccggacccgg caccgacggc ctccgcgccc tccctgcccc cctggcccct ctccgccagg 7800
tcggcggagg ccctgcgggc ccaggcccgt aggttgctgg cgtacgtggc cgagcacccg 7860
gatgtcgacc ccgccgacgt ggggtactcc ctcgcgcgcg gacgggccgt gttcgagcac 7920
cgggccgtgc tcctcggcac cggccacgac gacttccggc gcgccctgga cgccctggcg 7980
tcgggcgcgc ccgacggcgc ggtcgtccag ggcgcggcgg tggggcggca gggcaaggtc 8040
gtctttgtgt gctcggggca gggcacccag cgccccggca tgggccgcgg gctctaccgc 8100
tcgtccacgg cgttcgccgg ggcgctggag gaggtgtgcg cccatctgga cccgtatctg 8160
gaacaccctc tgatggaggt gatgttcgcc gatgagaaga gcgatacgtc ggcgctgctg 8220
catctgaccg cctacgccca accggccctc ttcgccctcc agaccgcgct gcaccgcatg 8280
gtcaccgagg agttcgggct cacccccgac tatctggccg gccactccct gggcgagctg 8340
accgccgccc atctggcggg catcctcagc ctgcccgacg ccgcggcgct ggtggcggcc 8400
cgcgcccgcg ccatgcggga ccttccagcg accggagcca tggtcgccgt cgaggccacc 8460
gaggcggagc tgcggccccg gctcgccgag ttggcggacc gggtcggcat cgccgccgtc 8520
aacgcccccg cgtccctggt catcaccggc gaccacgacg ccgtgcacca gatcgccgac 8580
gacttccgcg ggcagggcag gaaggtcact cccctccagg tcagcggcgc cttccactct 8640
ccccatatgg agcccctgct cgacgagatc gggcgcaccg ccgaaaccct cacctaccac 8700
cggccccaca ctcccctcgt caccgcgtcg gccgacggcg gcgacgacac gaccgagccg 8760
cgggccgacg acgacccggg cacggccgcg ttctggcctc tccaggcccg gcgcaccgtc 8820
cactacgcgc gggccgtgga gcggctgcgc gcccgcggcg tcaccacgtt cctggaactc 8880
ggccccgact ccaccctcac taccctcgtc caccacaatc tcgccgcgca cgatcccgtg 8940
gccgtctccc tgctccatcc ggagcggtgc gagacgcaca gcgtcctcgg cgcactcgcc 9000
gcggtccacg cccacagccg ccccgtcgac tggacacgcc actacaccgc acggccgcgg 9060
ccgacgccac accagatcga cgtgcccacc tatgccttcc ggcaccggcg ctactggctg 9120
cccgccccgg cggcggtcgg cgatgtgacg gccgcggggc tcgacgcggc ggagcacccg 9180
ctgatcggcg ccgccgtgtg gctcgcggag ggcgacggct gtctgctgac cggcaggatc 9240
tcgccgcgta cgcacccgtg gctggccgac catgtcatcg ccggcactgt gctccttccg 9300
ggcaccgcgt tcgtggagct ggcgctgcgg gccggggcgt acgtgggctg cgaccgtgtg 9360
gaggagctga ccctgcacgc gccactcccg ctgcccgccg acggtgaggt ggtgctgcag 9420
gtggcggtgg gggccgccga cgagtccggc cgccgtgagc tgagcatcca cgcccggccg 9480
gcggacgacg gtacatggac acggcacgcc atcggcacgc tggcatcggc ccgcggcgtc 9540
ggcctcgacg atggcacggg gcacaatggc cacgccccgg cgggcgacga gccgttcggg 9600
tcgtgggcca cggcctggcc gccgcccggt gccgagccct tggacgtcac cggggtctac 9660
gaccggtttg ccgacgccga gttcacgtac ggggaggcat tccaggggct ggtcgcggct 9720
tggcggcacg gcgacgagac gctggcggag gtccgcctcc ccgaccagcc ggccggtgac 9780
gccctccgct tcgggctgca ccccgcgctg ctcgacgcgg cactgcagac catgtggctc 9840
gtggagcccg acggcacacg gccgagcggt ggcctgggcg gccccgatcg gggcctgccg 9900
ttcgcctggc agggggtctc gctgcgtacg gcgggcccgt cggccctgcg ggtacggctg 9960
cgacggccgg cgccggacac cgtggccgtc gccgtggccg acgcggccgg ccggccggtc 10020
gcgtcggtgg agtcgctgac gctgcggccg gtgccgcggg gcgccttgcg cggcaccgag 10080
acggcggtgc gcacctcgtt gtacggcctg gactggacgg atgtgccgct gccgacgccg 10140
cagacggccc tgccccggtg tgcgctgatc ggagcggaca cgctcgacct ggtccccgcg 10200
ctcgaggccg cggcgcccga ccgcatcacc gacggcgtgg agcgctacgc cgacctggag 10260
gagctggtgc gctccgtggc ggcgggcgcc cccgccccgg acctcgtcat cgccggctgc 10320
cacgcagccc ctgaagccga cggcgcgagc gaacagccac agcccgagac ggtgcgcaca 10380
aggacgggtc aggtgctgga gctgcttcag cggtggctcg gcgcggacgg gctcgccgac 10440
gcacacctgg tgctgttcac ctcaggcgcg gtcgccaccc ggccgggcga gccggtgcgg 10500
gacctggcgg gggcggcggt ctggggtctg gtgcgctccg gccagtcgga gcatccggag 10560
tgcttcaccg tggtggacat ggacggcgcc caggagtccc gcgcggcgct gctcggcgcg 10620
ctcggcctcg gcgagccgca actggcggtg cgcggcggcc gggcgctggc gccgcgcctg 10680
gtgcgcccgg gtgacgccga cgacgacagc ggcctggccc tgccgcaggg gccggaaggc 10740
tggcggttgg agtgtcccgg cacgggcagc ctggacgggt tgaccacgac cgagtccccg 10800
gccgcggcgg tgccgctcgg cccgggcgag gtacgggtcg cggtgcgggc cgcggggctg 10860
aacttccgcg atgtgctgat cgcgctgggc gtggtgcccg ggcggacggc gctgggcagt 10920
gagggggcgg ggatcgtcct cgaggtcggg gcggaggtcc gcgatctcgc gcccggggac 10980
cgggtggtgg gtatcttccc cgaggcgttc ggcccggtgg ccgtggccga gcgggcgacc 11040
ctggcgcggg tccccgacgg ctggtcgttc gcccaggccg cgtcggtccc catcgtgttc 11100
gccaccgcgt accacggcct ggtcgatctg gcgcgcctgc ggccggggga atcggtgctg 11160
atccatgccg cggccggcgg ggtgggcatg gccgccgtgc aactggcgcg ccatctgggg 11220
gccgaggtgt acgccacggc cggccccggc aagtggcaca tcctgcgttc ccaaggcatc 11280
gacgacgacc atctggcctc gtcgcgcacg ctggagttcg agcagcgctt cgccgcgacc 11340
cgcggcgggc gggggatcga tgtcgtcctg gactgtctgg cccatgagtt cgtcgacgcc 11400
tcgctgcgcc tggtggcgcg tgacggcggc cggttcctgg agatgggcaa gagcgacatc 11460
cgtgacccgc ggcaggtggc gctggaccat ccgggcgtgc tctaccgggc gttcgacctg 11520
ctggaggccg ggccggagcg ggtcgggcag atcctgcgca ccgtactgga cctgttcgag 11580
cgcggtgtcc tggcgcacct gccgacgacc tgctgggaca tccggcaggc ggagcacgcc 11640
ttccgccatc tgcagcaggg ccgtcacatc ggaaagaacg tgctcaccgt cccggccggc 11700
tggaacgccg agggcaccgt actgatcacc ggcggtatgg gcaccctggg cgccgccctc 11760
gcccgtcatc tggcgggtac cgggcgcgcc cgccatctgc tgctggccgg ccgacgcggc 11820
cccgacgccc cgggcgccga ggagctgcga gaggagctga ccgagctggg cgcgcgggtc 11880
accatcgccg catgcgatct cggcgaccgg gcggcggtcg cccggctcct gggggcgatc 11940
ccggccgagc ggccgctgac cgctgtcatc cacgcggcgg gtgtcgtcga cgatgccacc 12000
ctcgggtccc tcaccccccg ccacctggac gccgccctgg ccgccaaggc cgacgccgcc 12060
tggcatctgc acaccctcac ccgccacgcc gacgtggccg cgttcgtcct cttctcctcg 12120
gtcgcgggtc tgctcggctc gcccgggcag ggcaactacg ccgcggccaa cgccttcttg 12180
gacgcgctcg cccaccaccg gcgcggctct ggccttccgg cggtgtcgct ggcgtggggg 12240
ctgtgggagc agaccagcgg catgaccggg cacctggacc aggccgaccg cgcccggctg 12300
gcccggctcg gcatcagccc gctcacgacc gggcaggcgc tcggcctttt cgacgccgcc 12360
ctcggccacc accgccccgt gctcgtcccc gcccgcctcg acgtgcccga tccgcacccc 12420
ggctcgtcga ccgtgccgcc cctgtaccgg ggcctggtcg gatccaggac ccggcggaca 12480
ccccccgcgg ccgccgccac cgggccgttc cccctgcata cccgcctcgg cggtcacgcc 12540
ccggccgagc agcacgagat gctgctctcg ctggtccgct cccacgccgc cctcgtgctg 12600
ggccgcgacg atccggacac ggtccatccc ggcgcgcact tccgcggcct gggcttcgac 12660
tccctgaccg cggtcgagct ccgcaaccgg ctcaacgccg ccaccggcct ccggctctcc 12720
accaccctcg tcttcgacca ccccacgccc gacgaactcg cccgtcacgt ccgggagcag 12780
gtgctgggcg acggcgaagc ggcgcgggtg gccccggtgc tggccgagct cgacaggctg 12840
gaagcggcgc tgtcccgggt ggacggggac gatgcggtcc gggcgagggt gacggcccgg 12900
ttgcaggccc ttctcctgaa gtggaacgag tccgatggtc cggcgacggg cggtgacggt 12960
gcgggcaggc tggcgtccgc cacggccgcc gaggtgctgg atttcatcag gaacgacctc 13020
ggcctctcct ga 13032
<210> 71
<211> 16599
<212> DNA
<213> Artificial Sequence
<220>
<223> aveA3 gene of Streptomyces avermitilis MA-4680
<400> 71
atggacacgt ccagcgaaaa gctcgtcgac gcgcttaggg cgtctctgaa ggcgaaccag 60
accctgcggg cacgtaatga gcaactggca gccgccatgg aggcgtccag cgagccgatt 120
gcgattgtgg ggatggcgtg tcgttttccg ggtggggtgt gttcgccgga ggagttgtgg 180
gagctggttg cgtcgggtgg ggatgcgatt ggtgaatttc cggccggtcg ggggtgggat 240
ctggaggggt tgtttgattc ggaccctgac cggtcgggga cgtcgtacgc gcggtatggc 300
gggtttttgt atgaggcggg ggagttcgat gcggacttct tcgggatcag tccgcgtgag 360
gcgttggcga tggatccgca gcagcggttg ttgctggaga cgtcgtggga ggcgttcgag 420
cgggcgggta tcgatccgct gtcgatgcgt ggctcccgta cgggtgtctt cgccggggtg 480
atgtaccacg actacggatc ccgcctgggt accatccccg agggattcga gggctacatc 540
ggcaacggta gcggcggcgc cgtcgcgtcg ggccgcgtcg cctacacgct cggtctcgag 600
ggccctgccg tctcggtgga cacggcatgt tcgtcgtcgt tggtggcgct gcatctggcg 660
tgccagtcgc tgcggtcggg tgagtgcacg ctcgcgctgg ccggcggtgt gacggtgatg 720
tcgaccccgc acctcttcgt cgagttctca cgccagcgcg gactgtcggt ggacggccgc 780
tgcaagtcct tcgcgggtgg agccgacggc accggcatgg gcgagggcgt cgggatgctg 840
ttggtggagc ggttgtcgga tgcggtgcgg ctggggcatc gggtgctggc ggtgctgcgc 900
ggcagtgcgg tcaatcagga cggtgcgtcg aatgggttga cggcgccgaa tggtccggct 960
caggagcggg tgatccggca ggcgttggcg aacgcggggt tgtccgtggc ggatgtggat 1020
gtggtggagg ggcatgggac gggcacgacg ctgggtgatc cgatcgaggc gcaggcgttg 1080
ctcgccacgt acgggcagcg ggccggtaac aggccgctgt ggctgggatc ggtgaagtcg 1140
aacatcggcc atgcgcaggc tgccgcgggt gtgggtgggg tcatcaagat ggtgatggcg 1200
ttgcgggagg gggtgttgcc gcggacgttg catgtggatg agccgtcgcc gcaggtggac 1260
tggtccgcgg gggcggtgcg gctgctgacg gaggcggtgc cgtggccggg ggacgcggca 1320
gggcggttgc ggcgggcggg agtgtcgtcg ttcggggtca gtggcacgaa tgcgcatgtg 1380
attttggagg aggcgccggc ggcggggggc tgtgttgccg ggggtggggt gttggagggt 1440
gctccgggtc ttgccatttc ggtggctgag tcggtggccg ctccagtggc tgtgtctgcg 1500
ccggtggctg agtcggtgcc ggtgccggtg ccggtgccgg ttcctgtgcc ggtgtcggct 1560
aggtctgagg ctgggttgcg ggcgcaggcg gaggcgttgc gtcagtacgt ggcagtccgg 1620
ccggacgttt cgcttgccga tgtgggtgcg ggtctggcct gtgggcgggc tgtgctggag 1680
catcgtgcgg tcgtcctggc cgcggaccgt gaggagctgg tgcaagggtt gggggcgctg 1740
gcggcgggtg agccggatcg gcgggtgacc acgggtcatg cgccgggtgg tgaccggggc 1800
ggtgtcgtct tcgtgtttcc cggacagggt gggcagtggg ccgggatggg tgtgcgtctg 1860
ctcgcctcct ctccggtgtt cgcccggcgg atgcaggcgt gcgaggaggc tctggcgccg 1920
tgggtggact ggtctgtggt ggacatcctg cgccgggacg cgggggatgc ggtgtgggag 1980
cgggccgatg tggtccagcc tgtgctgttc agcgtcatgg tgtctttggc tgctctgtgg 2040
cgttcctacg gtatcgaacc cgacgcggtc cttggccatt cccagggcga gatcgcggcc 2100
gcgcatgtgt gtggggcgct gagcctgaag gacgcggcga agactgttgc gctgcgcagc 2160
cgggcgctgg ccgctgtgcg gggccggggc ggcatggcct cagtgccgct gcctgcccag 2220
gaggtggagc agctcattgg tgagcggtgg gcggggcggt tgtgggtggc ggcggtcaac 2280
ggcccccgct ccaccgccgt ctcgggggat gccgaggcgg tggacgaggt gctggcgtac 2340
tgtgccggca ccggggtgcg ggcccggcgg atcccggtcg actatgcctc gcactgcccc 2400
catgtgcagc ccctgcggga ggagttgctg gagctgctgg gggacatcag cccgcagccg 2460
tccggcgtgc cgttcttctc cacggtggag ggcacctggc tggacaccac aaccctggac 2520
gccgcctact ggtaccgcaa cctgcaccag cctgtccgtt tcagcgatgc cgtccaggcc 2580
ctggcggatg acggacaccg cgtcttcgtc gaagtcagcc cccaccccac cctcgtcccc 2640
gccatcgaag acaccaccga agacaccgcc gaagacgtca ccgcgatcgg cagcctccgc 2700
cgcggcgaca acgacacccg ccgcttcctc accgccctcg cccacaccca caccaccggc 2760
atcggcacac ccaccacctg gcaccaccac tacacccacc accacaccca cccccacaac 2820
caccacctcg acctccccac ttatcccttc caacgccagc actactggct cgacgctccc 2880
acgggagcag gtgacgtcgc cgctgctggc ttggagccgg ccgaacaccc tctgctcgcg 2940
gcaacagtcc aactcgcaga cacggacggc tgcctactga cgggtcgcct gtccttgcgc 3000
tcgcatccgt ggctgggcga ttacgaggtg gggggtgcgg tcctgctgtc ggggtcggcg 3060
ttcgtggagc tggcggtcca ggttggcgaa cgcgtgggct gcacccgaat cgagcaactc 3120
actgtgcatg cgccgctggt ggttcctgtg ggtgggggtg tgagtgtgca ggttggggtt 3180
gcggctgcgg atggggaggg gcggcgtttg gtgagtgtgt atgcgcgggg tgggagtgct 3240
tgtggtgggg gtggtgcgtc gggtggggtg tggacgtgtc atgcctcggg ggtgctggtt 3300
gaggctgctg ctggtggtgg tgtggtggtg gatggtctgg cgggggtgtg gccgccgcgg 3360
ggtgcggtgg cggtggatgt cgatggtgtc cgtgaccgtt tggctggggc tggttgtgtt 3420
ttggggccgg tgttttcggg gctgcgtgcg gtgtggcgtg atggggggga tttgctggct 3480
gaggtgtgtc tgccggagga ggcgtggggt gatgcggctg gttttgggct gcatccggcg 3540
ttgctggatg gtgtggtcca gccgttgtcg gtgttgcttc cgggtgggac ggggtttggg 3600
gagggggcgg ggttcgggga gggtgttcgg gtgccggctg tgtggggtgg tgtgtcgctt 3660
caccgggcgg gtgtgaccgg tgtgcgggtg cgtgtgtggg ctgtagggcg gggcggcggg 3720
cgtgaggcgg tgtcggtcgt ggtcggggat gaggcgggtg tgccggtggc gtcggtcgat 3780
cgtcttgagt tgcggcctgt ggatatgggt cagttgcgtg ctgtctcggt ttcggcgggg 3840
cggcggggtt cgctgtatgc ggtgcagtgg gctgaggtgg gtcctgtgcc ggtgtgtggg 3900
caggcgtggg cgtggcacga ggacgtgggt gagagcggtg gtgggcctgt gccgggggtg 3960
gtggtgttgc ggtgcccgga tgccggtgcc ggtggcggcg gtggcggtgg tgtgggtgag 4020
gttgttggtg gggtgttggg tgtggtgcag gggtggctgg ggctggagcg gtttgcgggt 4080
tcgcggctgg tggtggtgac ccggggtgcg gtggtggccg gccaagaaga cggcccggtg 4140
gatgtggtgg gtgcggcggt gtgggggctg gtgcggtcgg cgcaggctga gcatccggac 4200
cggtttgtcc tcctcgacct cgacaccgac accgacaccg gcaccgacct cgacaccggt 4260
gctggtgctg gtgctggtgc tggttggggc gtggatggtg ggcatgtggc ggcggtggtg 4320
gcgtgtggtg agccgcagtt ggcggtgcgt ggtgagcggg tgctggccgc acgcctgacg 4380
cgacttgagt cgtccgttga tgtacctgct cagcggtccg gtgatgttgc tggtcgggag 4440
gtgttgccgt ggttgtcggg tgggtcggtg ttggtgacgg gtgggacggg tgtgctgggt 4500
gcggcggtgg cgcggcatct ggctggtgtg tgtggggtgc gggatctgct gttggtgagc 4560
cggcgtggtc cggatgctcc gggtgcggag ggtttgcggg cggagctggc cgcgttgggg 4620
gcggaggtgc ggattgttgc gtgtgatgtg ggggagcggc gggaggtggt ccggctgctg 4680
gagggtgttc ctgccgggtg tccgctgacg ggtgtcgtgc atgcggctgg tgtgctggac 4740
gatgcgacga tcgcctctct cacgcccgag cggctgggca cggtgttcgc ggccaaggtg 4800
gatgccgctc ttttgctgga tgagctgacg cggggtatgg agctgtcggc gttcgtgctg 4860
ttctcctcgg ccgcggggat cctggggtcg gccgggcagg gcaactacgc cgcggccaat 4920
gccgctctgg acgcgctggc gtaccggcgg cgggcggcgg gtctgccggg ggtgtcgctg 4980
gcgtgggggc tgtgggaaga ggccagcggg atgaccgggc acctggccgg caccgaccac 5040
cggcgcatca tccgttccgg tctgcatccc atgtcgaccc cggacgcact ggctctcttc 5100
gatgcggccc tggctctgga ccggccggtc ctgctgcccg ccgacctgcg tcccgccccg 5160
cccctgccgc ccctgctgca ggacctcctg cccgccaccc gccgccgcac cacccgcacc 5220
accactaccg gtggtgcgga caacggcgcc cagctgcatg cccggctggc cggccagaca 5280
cacgaacaac agcacaccac cctcctcgcc ctggtccgct cccacatcgc caccgtcctc 5340
ggccacacca cccccgacac catccccccc gaccgcgcgt tccgcgacct cggcttcgac 5400
tccctcaccg ccgtcgaact acgcaaccgg ctctcccgca ccaccggact ccgcctcccc 5460
accaccctcg ccttcgacca ccccaacccc accaccctca cccaccacct ccacacacaa 5520
cttctgggct cggacagcac tgcctccatc ccagctcccc gtgctgcggc tgtgcctgca 5580
gaccaggacg agcccgtcgc gatcattggc atggcgtgcc gctatcccgg aggcgtcacc 5640
tcagccgagg agctgtggga actgctcgca tcggggaggg acacggtcgg cgagtttccg 5700
acggaccgtg ggtgggacct ggaagcactg ttcgatccgg aaccgggtcg gccgggcacc 5760
tcgtacaccc gctgtgggag tttcctctac gacgcggggg agttcgacgc cggcttcttc 5820
gggatcagtc cgcgtgaggc actggcgatg gacccgcagc agcgattgct gctggaggcc 5880
tcatgggagg ccatggagca ggcaggtatt gaccctacga ccgtacgcgg gagccagaca 5940
ggcgtgttcg cgggcctcat tccgcaggcc tatggaccca ggctgcacga aaacgccgca 6000
gccgacaccg agggctatgt cctgaccggc acatccggga gtgtggcctc cggtcgtatc 6060
tcgtacacgt ttggttttga gggtcctgcg gtgtcggtgg acacggcttg ttcctcgtcg 6120
ttggtggctt tacatctggc ctgtcaggcg ttgcgtgcgg gtgagtgctc gatggcgctt 6180
gccgggggtg tgacggtgat gtcgtctccg ggtgccttcg tggagttttc gcggcagcgg 6240
ggtctggccg cggacgggca ttgcaaggcg ttctcggcgg cggcggacgg gaccggctgg 6300
ggtgagggtg tggggatgct gctggtggag cggctctccg acgcccgtcg caacggtcac 6360
cgtgtcctgg ccgtggtgcg tggcagtgcg gtcaaccagg acggtgcgag caacgggctg 6420
accgcgccca acgggccctc ccagcagcgt gtcatccgcc aggccctcgc caacgccggc 6480
ttgtcggccg gtgatgtcga tgcggtggag gcccacggca ccggcaccac tttgggcgac 6540
ccgatcgagg cccaggccct ccttgcgacc tacgggcagg accgtgccgg cgaggggccg 6600
ctgtggctgg gctcggtcaa gtccaatgtc ggtcacacac aggctgccgc gggcgtcgcc 6660
ggggtgatca agatggtgat ggcgctgcgg aatggtctgc tgccgcggac gttgcatgtg 6720
gatgagccgt cgccgcatgt ggactggtcc gcgggtgcgg tgcagctgct gacggagacg 6780
gtgccctggc ccggcgggga ggggcggcta cggcgggcag gagtgtcatc attcggcgtc 6840
agcggcacca acgcccacgt catcctcgaa gaagcacccg cccacaacat cccgtcagac 6900
acacccgccg acgacgttcc ggggggacca cccgccggcg aggatgccgg tagtggcgag 6960
gaggctgctg ccggcagtcc aggggtgtgg ccgtggctgg tgtcggccaa gtcgcagccg 7020
gccctgcgcg cccaggccca ggccctgcac gcccacctca ccgaccaccc cggcctcgac 7080
ctcgccgacg tcggatacac cctcgcccac gcccgcgccg tgttcgacca ccgcgccacc 7140
ctcatcgccg ccgaccgcga caccttcctg caagcactcc aggcactcgc cgcaggcgaa 7200
ccccaccccg ccgtcatcca cagcagcgcc ccaggcggga ccgggaccgg ggaggccgca 7260
ggaaagaccg cattcatctg ctccggacag ggcacccaac gccccggcat ggcccacggc 7320
ctctaccaca cccaccccgt cttcgccgcc gcactcaacg acatctgcac ccacctcgac 7380
ccccacctcg accaccccct cctccccctc ctcacccagg accccaacac ccaggacacc 7440
accaccctcg aagaagcggc cgcactgctc cagcagaccc cgtacgccca gcccgccctc 7500
ttcgccttcc aggtcgccct ccaccgcctc ctcaccgacg gctaccacat caccccccac 7560
tactacgccg gacactccct cggcgaaatc accgccgccc acctcgccgg catcctcacc 7620
ctcaccgacg ccaccaccct catcacccaa cgcgccaccc tcatgcaaac catgcccccc 7680
ggcaccatga ccaccctcca caccaccccc caccacatca cccaccacat caccgcccac 7740
gaaaacgacc tcgccatcgc cgccatcaac acccccacct ccctcgtcat cagcggcacc 7800
ccccacaccg tccaacacat caccaccctc tgccaacaac aaggcatcaa aaccaaaacc 7860
ctccccacca accacgcctt ccactccccc cacaccaacc ccatcctcaa ccaactccac 7920
cagcacaccc aaaccctcac ctaccaccca ccccacaccc ccctcatcac cgccaacacc 7980
ccacccgacc aactcctcac cccccactac tggacccaac aagcccgcaa caccgtcgac 8040
atagccacca ccacccaaac cctccaccaa cacggcgtca ccacctacat cgaactcgga 8100
cccgacaaca ccctcaccac cctcacccac cacaacctcc ccaacacccc caccaccacc 8160
ctcaccctca cccaccccca ccaccacccc caaacccacc tcctcaccaa cctcgccaaa 8220
accaccacca cctggcaccc ccaccactac acccaccacc acaaccaacc ccacacccac 8280
acccacctcg acctccccac ctaccccttc caacaccacc actactggct cgaaagcaca 8340
cagcccggtg ccggcaacgt gtcagcagcc ggactcgacc ccaccgaaca ccccctactc 8400
ggcgccacat tggaactggc cgaaggggac ggctgcctac tgacggggcg cctctcgttg 8460
cgcacgcatc cctggctcgc cggccatgcg gtaggcggtg tcgtgctgct gccgggtacg 8520
gccttcgcgg aactggccct tcatgccgga gaaagtgtgg gttgcgacca cgtggacgag 8580
ctgacgctcc acacaccgtt ggtcattcct gaggtcggag acgtgaccct tcaggttgcc 8640
attgcggcgc cggacgagtc gggtcgccgc atgatgacca tccactcacg cggtgagggc 8700
ggcagtggtg gagccgatgc gtcggccagt gcgtggacgc gtcatgccgc gggtgtgctg 8760
agccctgcca aggacgatga cactgcctcg tacgagctgc ttgcgggacc ctggcctccc 8820
gttggagcta cgcctgtcga cctgaacacg gcttacgatc aaatggccga cgccggcttt 8880
gcttatggcc tggcattcca agggttgcgc gcggcctggc gctacggcga cgacatcctc 8940
gtcgaggcac gtcttcccga agaagtgtcg ggagacgcgg cggcgtacgg tctgcacccg 9000
gccctgctcg acgctgccct tcagggcacc ggcctgcttt ctgtggcggg tccggggacg 9060
cccgtcgtgc cccatgtgtg gaacggtctg cggttccgta cgcatggtgc agtctccgtg 9120
cgcgcgtgcc tgtcgacgct tggagcgaca ggggcggccg tgtgcgtgcg catcaccgac 9180
gacaccgggg tgccggtggc gtcggtcgat cgtcttgagt tgcggcctgt ggatatgggt 9240
cagttgcgtg ctgtctcggt ttcggcgggg cggcggggtt cgctgtatgc ggtgcagtgg 9300
gctgaggtgg gtcctgtgcc ggtgtgtggg caggcgtggg cgtggcacga ggacgtgggt 9360
gagagcggtg gtgggcctgt gccgggggtg gtggtgttgc ggtgcccgga tgccggtgcc 9420
gatggcggcg gtggcggtgg tgtgggtgag gttgttggtg gggtgttggg tgtggtgcag 9480
gggtggctgg ggctggagcg gtttgcgggt tcgcggctgg tggtggtgac ccggggtgcg 9540
gtggtggccg gcccggagga cggcccggtg gatgtggtgg gtgcggcggt gtgggggctg 9600
gtgcggtcgg cgcaggctga gcatccggac cggtttgtcc tcctcgacct ggacaccgac 9660
ctcgacagcg gcgctgacgc cgatgccggc aacgaggccg gtatggggtc tggtctggat 9720
ggtgggcgtg tggctgcggt ggtggcgtgt ggtgagccgc agttggcggt gcgtggtgag 9780
cgggtgctgg ccgcacgcct gacacgactt gagtcgccgg ttgatgtatc gggtcgggag 9840
gtgttgccgt ggttgtcggg tgggtcggtg ttggtgacgg gtgggacggg tgtgctgggt 9900
gcggcggtgg cgcggcatct ggctggtgtg tgtggggtgc gggatctgtt gttggtgagc 9960
cggcgtggtc cggatgctcc gggtgcggag ggtttgcggg cggagctggc cgcgttgggg 10020
gcggaggtgc ggattgttgc gtgtgatgtg ggggagcggc gggaggtggt ccggctgctg 10080
gagggtgttc ctgccgggtg tccgctgacg ggtgtcgtgc atgcggctgg tgtgctggac 10140
gatgcgacga tcgcctctct cacgcccgag cggctgggca cggtgttcgc ggccaaggtg 10200
gatgccgctc ttttgctgga tgagctgacg cggggtatgg agctgtcggc gttcgtgctg 10260
ttctcctcgg ccgcggggat cctggggtcg gccgggcagg gcaactacgc cgcggccaat 10320
gccgctctgg acgcgctggc gtaccggcgg cgggcggcgg gtctgccggg ggtgtcgctg 10380
gcgtgggggc tgtgggaaga ggccagcggg atgaccgggc acctggccgg caccgaccac 10440
cggcgcatca tccgttccgg tctgcatccc atgtcgaccc cggacgcact ggctctcttc 10500
gatgcggccc tggctctgga ccggccggtc ctgctgcccg ccgacctgcg tcccgccccg 10560
cccctgccgc ccctgctgca ggacctcctg cccgccaccc gccgccgcac cacccgcacc 10620
accactaccg gtggtgcgga caacggcgcc cagctgcatg cccggctggc cggccagaca 10680
cacgaacaac agcacaccac cctcctcgcc ctggtccgct cccacatcgc caccgtcctc 10740
ggccacaacg cgccggagat gatccccgtt gactcggcgt tccgcgacct aggcttcgac 10800
tccttgacag cggtggaact ccgtaaccgc ctgggtgagg caacgggact gcgactgccg 10860
accagtctgg tcttcgacca gccgaatgca gcgaccctgg cgcgtcacct acgtcgtgag 10920
ctgatgggcg acgacgcgga aggcgagacg ccatcgcagg tcgcacttca tcaggttgcc 10980
gcggatgagc cgattgcgat tgtggggatg gcgtgtcgtt ttccgggtgg ggtgtgttcg 11040
ccggaggagt tgtgggagct ggttgcgtcg ggtggggatg cgattggtga atttccggcc 11100
ggtcgggggt gggatctgga ggggttgttt gattcggacc ctgaccggtc ggggacgtcg 11160
tacgcgcggt atggcgggtt tttgtatgag gcgggggagt tcgatgcgga cttcttcggg 11220
atcagtccgc gtgaggcgtt ggcgatggat ccgcagcagc ggttgttgct ggagacgtcg 11280
tgggaggcgt tcgagcgggc gggtatcgat ccgctgtcga tgcgtggctc ccgtacgggt 11340
gtcttcgccg gggtgatgta ccacgactac gccgcgcgtc tccaccatgt ccccgagggt 11400
ttcgaaggcc tcatcgccaa cggcagcgca ggcagcgtcg cgaccggccg ggtggcctac 11460
agctttggcc ttgagggtcc ggccgtgacc gtcgatacgg cgtgttcgtc gtcgttggtg 11520
gcgttgcatt gggcggcgca ggcgttgcgt gcgggtgagt gttcgatggc gcttgccggg 11580
ggtgtgacgg tgatgtcgtc tccgggtacg tttgtggagt tctcacgtca gcggggtctg 11640
gccgcggacg ggcggtgcaa ggcctattcg gcggctgctg acggtaccgg ctgggccgag 11700
ggtgtgggga tgctgctggt ggagcggctc tccgacgccc gtcgcaacgg tcaccgtgtc 11760
ctggccgtgg tgcgtggcag tgcggtcaac caggacggtg cgagcaacgg tctgaccgcg 11820
cccaacgggc cctcccagca gcgtgtcatc cgtcaggccc tggccaatgc gggactgacc 11880
ccggccgatg tcgacgcagt ggagggccac ggcaccggga ccactctggg ggacccgatc 11940
gaggcccagg cactcctggc cgcctacgga caacaccgcc cccaccaccg ccccttgtgg 12000
ctgggatccc tcaaatccaa catcgggcac gcacaggccg ccgcgggcgt gggcggagtc 12060
atcaagatgg tgatggccct gcgcaacggg ctgctgccac agaccctcca cgtggacgag 12120
cccacccccc aggtcgactg gtccacaggc gcagtacaac tcctgacaca accggtgccc 12180
tggcccgccg acccggccgg ccggccacgc cacgccggcg tgtcatcatt cggcgtcagc 12240
ggcaccaacg cccatgtgat tttggaggag gcgcctgcgg cggcgggcgg tgctgccggt 12300
ggtggggtgt cggtgggtgc tccgaatcca gcccttccgg tggctgagtc tgagccggtg 12360
ccggtgccgg tgccggtgtc ggcgaggtct gaggccgggt tgcgggcgca ggcacaggcg 12420
ttgcgccagt acgtggcagc ccgcccggac atgtcacctg ccgacatcgg tgcgggtctg 12480
gcccgcggcc gggccgtact ggaacaccgc gccgtcatcc tggccgcgga ccgcgaggaa 12540
ctggcgcagg cactgacagc cctggcagcc ggcgaacccc acccccacat caccacaggc 12600
cacacccggg gcagtgaccg cggcggcgtc gtcttcgtct tccccggaca gggcggccag 12660
tgggccggga tgggcctgac cctgctcacc tcctcacccg tgttcgccga acacatcgac 12720
gcatgcgaga aagccctcac cccctgggtg ccctggtccc tgaccgacat cctgcaccgc 12780
gaccccgacg accccgcatg gcaacaagcc gacgtggtcc agcccgtgct cttcagcatc 12840
atggtctccc tcgccgccct gtggcgctcc tacggcatcg aacccgacgc ggtcctcggc 12900
cactcccagg gagaaatcgc cgccgcccac atctgcggcg cactcagcct gaaagacgcc 12960
gccaaaaccg ttgcactgcg cagccaggca ctggccgccg tacgaggccg gggcgccatg 13020
gtctcactgc ccctgcccgc ccaggacgtg cagcagctca tttccgaacg gtgggaaggg 13080
cagttgtggg tggcagccct caacggcccc cactccacca ccgtctccgg cgacaccacc 13140
gcagtagaag aactcctcac ccactgtgcc gacaccggcc tacgggccaa acgcatcccc 13200
gtcgactacg cctcccactg cccccacgtc caacccctcc acgacgaact cctgcacctg 13260
ctgggagaca tcacccccca gccgtccacc atgccgttct tctccaccgt cgtagggcac 13320
ctggtctggt acaccacaac cctggacgcc gcctactggt accgcaacct ccaccagccc 13380
gtccgcttca gccacgccat ccagaccctg accgacgacg gacaccgccc cttcatcgaa 13440
atcagtcccc accccaccct cgtccccgcc atcgaagaca ccaccgaaaa caccaccgaa 13500
aacatcaccg cgaccggcag cctccgccgc ggcgacaacg acacccaccg cttcctcacc 13560
gccctcgccc acacccacac caccggcatt cggacaccca ccacctggca ccaccactac 13620
acccaaaccc acccccaccc ccacaaccac cacctcgacc tgcccaccta ccccttccaa 13680
caccagcact actggctcca accacccacc acgacaaccg acctcaccac caccggcctc 13740
acccccaccc accaccccct cctcaccgca acactcaccc tcgccaacaa caacacacaa 13800
ctactcaccg gccgcctctc cctacgcacc cacccctggc tcaccgacca caccgtcgtc 13860
ggtaccactc ttgtgccagg aaccgccctc ctcgaactcg ccctccaagc aaccacgacc 13920
gaccacctcg aagaactcgc cctccacacg cctctcgtca tcccccgtga gggtgccgtc 13980
gacgttcagg tgcacatcaa tccaccggac gacaccgaca ctcgttcact gacgatctac 14040
tcgcgaagcg agaacgcccc cgcagcggct ccctggcgtc atcacgccac ggccgttctg 14100
ggaaccaaga cctcgcgcat tgagacaggc cgtagccacg atgatctgtc gatgtggccg 14160
ccagcgggcg cagttcgctg tgctgatgag gaattggcag ccttgtatgg cgactacgag 14220
gcaaatggct ttgtctatgg ccccgcattc cgggggctga ctgctgcctg gcgtctggga 14280
gacgaggtgt ttgccgaggt tcgccttcca gaacaggtgc acggcgaggc atccgcgtac 14340
aacctgcacc cggcactgct ggatgctgcc ttgcacgcag cggcctttgc gccgtcgggc 14400
agtctgccgc agggatccgt accgttctcc ttcaccggtg tgacgctgca cgccgccaat 14460
gcgtcgtcgt tgcgcgtgcg actctcgccg gccgatccga acagcggcca cgccgcagtt 14520
tccgtgctgg tcacggatga caccggtacg cccgtggcgt ccgtcgaggc gttggcggtg 14580
cgcccgttgg cggcggacga attgcgagct gccgagcgcg ccgtacagcg cgctgagctc 14640
ttcgacatga agtgggttga ggtgccctca gatgtactgg tgtcgggcgg ggcatcggtg 14700
gtggtgctgg atggtgccga cgacctcgtt ggtctggcgg ctgaggagga tggtgtgccg 14760
ggggtggtgg tgttgcggtg cccggatgcc ggtgccgatg gcggcggtgg tggcggtggt 14820
gtgggtgagg ttgttggtgg ggtgttgggt gtggtgcagg ggtggctggg gctggagcgg 14880
tttgcgggtt cgcggctggt ggtggtgacc cggggtgcgg tggtggccgg cccggaggac 14940
ggcccggtgg atggcccggt ggatgtggtg ggtgcggcgg tgtgggggct ggtgcggtcg 15000
gcgcaggctg agcatccgga ccggtttgtc ctcctcgacc tggacaccga cctcgacagc 15060
ggcgctgacc gcgatgccgg caacgaggcc ggtatggggt ctggtctgga tggtgggcgt 15120
gtggctgcgg tggtggcgtg tggtgagccg cagttggcgg tgcgtggtga gcgggtgctg 15180
gccgcacgcc tgacacgact tgagtcgccg gttgatgtat cgggtcggga ggtgttgccg 15240
tggttgtcgg gtgggtcggt gttggtgacg ggtgggacgg gtgtgctggg tgcggcggtg 15300
gcgcggcatc tggctggtgt gtgtggggtg cgggatctgt tgttggtgag ccggcgtggt 15360
ccggatgctc cgggtgcgga gggtttgcgg gcggagctgg ccgcgttggg ggcggaggtg 15420
cggattgttg cgtgtgatgt gggggagcgg cgggaggtgg tccggctgct ggagggtgtt 15480
cctgccgggt gtccgctgac gggtgtcgtg catgcggctg gtgtgctgga cgatgcgacg 15540
atcgcctctc tcacgcccga gcggctgggc acggtgttcg cggccaaggt ggatgccgct 15600
cttttgctgg atgagctgac gcggggtatg gagctgtcgg cgttcgtgct gttctcctcg 15660
gccgcgggga tcctggggtc ggccgggcag ggcaactacg ccgcggccaa tgccgctctg 15720
gacgcgctgg cgtaccggcg gcgggcggcg ggtctgccgg gggtgtcgct ggcgtggggg 15780
ctgtgggaag aggccagcgg gatgaccggg catctggccg gcaccgacca ccggcgcatc 15840
atccgttccg gtctgcatcc catgtcgacc ccggacgcac tggccctctt cgatgcggcc 15900
ctggctctgg accggccggt cctgctgccc gccgacctgc gtcccgcccc gcccctgccg 15960
cccctgctgc aggacctcct gcccgccacc cgccgccgca ccacccgcac caccactacc 16020
ggtggtgcgg acaacggcgc ccagctgcac ggccggctgg ccggccagac acacgaacaa 16080
cagcacacca ccctcctcgc cctggtccgc tcccacatcg ccaccgtcct gggccacacc 16140
acccccgaca ccatcccccc cgaccgcgcg ttccgcgacc tcggcttcga ctccctcacc 16200
gccgtcgaac tacgcaaccg gctctcccac accaccggac tccgcctccc caccaccctc 16260
gccttcgacc accccaaccc caccaccctc acccaccacc tccacacaca actcgtcagc 16320
aagggactca ccgccgcggc cgagccggac gccgcaacga cacccccggg gctgccctcg 16380
ctgctctcgg agctcgagcg gctggaggcg gtagtgctct cctccaccac atcctccgct 16440
gccccgctgg acgacggcgc gcgcacgcgg ctggcctccc gactgcattc cctcgcccag 16500
aagttgaacg gcgacgacac cgcccccgac ctcgcagaga catcggacga ggagatgttc 16560
gctctcatcg acagggaagt cggattcgaa tctcaatga 16599
<210> 72
<211> 5532
<212> PRT
<213> Artificial Sequence
<220>
<223> type I polyketide synthase AVES 3 (BAA84478.1)
<400> 72
Met Asp Thr Ser Ser Glu Lys Leu Val Asp Ala Leu Arg Ala Ser Leu
1 5 10 15
Lys Ala Asn Gln Thr Leu Arg Ala Arg Asn Glu Gln Leu Ala Ala Ala
20 25 30
Met Glu Ala Ser Ser Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg
35 40 45
Phe Pro Gly Gly Val Cys Ser Pro Glu Glu Leu Trp Glu Leu Val Ala
50 55 60
Ser Gly Gly Asp Ala Ile Gly Glu Phe Pro Ala Gly Arg Gly Trp Asp
65 70 75 80
Leu Glu Gly Leu Phe Asp Ser Asp Pro Asp Arg Ser Gly Thr Ser Tyr
85 90 95
Ala Arg Tyr Gly Gly Phe Leu Tyr Glu Ala Gly Glu Phe Asp Ala Asp
100 105 110
Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln
115 120 125
Arg Leu Leu Leu Glu Thr Ser Trp Glu Ala Phe Glu Arg Ala Gly Ile
130 135 140
Asp Pro Leu Ser Met Arg Gly Ser Arg Thr Gly Val Phe Ala Gly Val
145 150 155 160
Met Tyr His Asp Tyr Gly Ser Arg Leu Gly Thr Ile Pro Glu Gly Phe
165 170 175
Glu Gly Tyr Ile Gly Asn Gly Ser Gly Gly Ala Val Ala Ser Gly Arg
180 185 190
Val Ala Tyr Thr Leu Gly Leu Glu Gly Pro Ala Val Ser Val Asp Thr
195 200 205
Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ser Leu
210 215 220
Arg Ser Gly Glu Cys Thr Leu Ala Leu Ala Gly Gly Val Thr Val Met
225 230 235 240
Ser Thr Pro His Leu Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ser
245 250 255
Val Asp Gly Arg Cys Lys Ser Phe Ala Gly Gly Ala Asp Gly Thr Gly
260 265 270
Met Gly Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala
275 280 285
Val Arg Leu Gly His Arg Val Leu Ala Val Leu Arg Gly Ser Ala Val
290 295 300
Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala
305 310 315 320
Gln Glu Arg Val Ile Arg Gln Ala Leu Ala Asn Ala Gly Leu Ser Val
325 330 335
Ala Asp Val Asp Val Val Glu Gly His Gly Thr Gly Thr Thr Leu Gly
340 345 350
Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Arg Ala
355 360 365
Gly Asn Arg Pro Leu Trp Leu Gly Ser Val Lys Ser Asn Ile Gly His
370 375 380
Ala Gln Ala Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Met Ala
385 390 395 400
Leu Arg Glu Gly Val Leu Pro Arg Thr Leu His Val Asp Glu Pro Ser
405 410 415
Pro Gln Val Asp Trp Ser Ala Gly Ala Val Arg Leu Leu Thr Glu Ala
420 425 430
Val Pro Trp Pro Gly Asp Ala Ala Gly Arg Leu Arg Arg Ala Gly Val
435 440 445
Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Glu
450 455 460
Ala Pro Ala Ala Gly Gly Cys Val Ala Gly Gly Gly Val Leu Glu Gly
465 470 475 480
Ala Pro Gly Leu Ala Ile Ser Val Ala Glu Ser Val Ala Ala Pro Val
485 490 495
Ala Val Ser Ala Pro Val Ala Glu Ser Val Pro Val Pro Val Pro Val
500 505 510
Pro Val Pro Val Pro Val Ser Ala Arg Ser Glu Ala Gly Leu Arg Ala
515 520 525
Gln Ala Glu Ala Leu Arg Gln Tyr Val Ala Val Arg Pro Asp Val Ser
530 535 540
Leu Ala Asp Val Gly Ala Gly Leu Ala Cys Gly Arg Ala Val Leu Glu
545 550 555 560
His Arg Ala Val Val Leu Ala Ala Asp Arg Glu Glu Leu Val Gln Gly
565 570 575
Leu Gly Ala Leu Ala Ala Gly Glu Pro Asp Arg Arg Val Thr Thr Gly
580 585 590
His Ala Pro Gly Gly Asp Arg Gly Gly Val Val Phe Val Phe Pro Gly
595 600 605
Gln Gly Gly Gln Trp Ala Gly Met Gly Val Arg Leu Leu Ala Ser Ser
610 615 620
Pro Val Phe Ala Arg Arg Met Gln Ala Cys Glu Glu Ala Leu Ala Pro
625 630 635 640
Trp Val Asp Trp Ser Val Val Asp Ile Leu Arg Arg Asp Ala Gly Asp
645 650 655
Ala Val Trp Glu Arg Ala Asp Val Val Gln Pro Val Leu Phe Ser Val
660 665 670
Met Val Ser Leu Ala Ala Leu Trp Arg Ser Tyr Gly Ile Glu Pro Asp
675 680 685
Ala Val Leu Gly His Ser Gln Gly Glu Ile Ala Ala Ala His Val Cys
690 695 700
Gly Ala Leu Ser Leu Lys Asp Ala Ala Lys Thr Val Ala Leu Arg Ser
705 710 715 720
Arg Ala Leu Ala Ala Val Arg Gly Arg Gly Gly Met Ala Ser Val Pro
725 730 735
Leu Pro Ala Gln Glu Val Glu Gln Leu Ile Gly Glu Arg Trp Ala Gly
740 745 750
Arg Leu Trp Val Ala Ala Val Asn Gly Pro Arg Ser Thr Ala Val Ser
755 760 765
Gly Asp Ala Glu Ala Val Asp Glu Val Leu Ala Tyr Cys Ala Gly Thr
770 775 780
Gly Val Arg Ala Arg Arg Ile Pro Val Asp Tyr Ala Ser His Cys Pro
785 790 795 800
His Val Gln Pro Leu Arg Glu Glu Leu Leu Glu Leu Leu Gly Asp Ile
805 810 815
Ser Pro Gln Pro Ser Gly Val Pro Phe Phe Ser Thr Val Glu Gly Thr
820 825 830
Trp Leu Asp Thr Thr Thr Leu Asp Ala Ala Tyr Trp Tyr Arg Asn Leu
835 840 845
His Gln Pro Val Arg Phe Ser Asp Ala Val Gln Ala Leu Ala Asp Asp
850 855 860
Gly His Arg Val Phe Val Glu Val Ser Pro His Pro Thr Leu Val Pro
865 870 875 880
Ala Ile Glu Asp Thr Thr Glu Asp Thr Ala Glu Asp Val Thr Ala Ile
885 890 895
Gly Ser Leu Arg Arg Gly Asp Asn Asp Thr Arg Arg Phe Leu Thr Ala
900 905 910
Leu Ala His Thr His Thr Thr Gly Ile Gly Thr Pro Thr Thr Trp His
915 920 925
His His Tyr Thr His His His Thr His Pro His Asn His His Leu Asp
930 935 940
Leu Pro Thr Tyr Pro Phe Gln Arg Gln His Tyr Trp Leu Asp Ala Pro
945 950 955 960
Thr Gly Ala Gly Asp Val Ala Ala Ala Gly Leu Glu Pro Ala Glu His
965 970 975
Pro Leu Leu Ala Ala Thr Val Gln Leu Ala Asp Thr Asp Gly Cys Leu
980 985 990
Leu Thr Gly Arg Leu Ser Leu Arg Ser His Pro Trp Leu Gly Asp Tyr
995 1000 1005
Glu Val Gly Gly Ala Val Leu Leu Ser Gly Ser Ala Phe Val Glu Leu
1010 1015 1020
Ala Val Gln Val Gly Glu Arg Val Gly Cys Thr Arg Ile Glu Gln Leu
1025 1030 1035 1040
Thr Val His Ala Pro Leu Val Val Pro Val Gly Gly Gly Val Ser Val
1045 1050 1055
Gln Val Gly Val Ala Ala Ala Asp Gly Glu Gly Arg Arg Leu Val Ser
1060 1065 1070
Val Tyr Ala Arg Gly Gly Ser Ala Cys Gly Gly Gly Gly Ala Ser Gly
1075 1080 1085
Gly Val Trp Thr Cys His Ala Ser Gly Val Leu Val Glu Ala Ala Ala
1090 1095 1100
Gly Gly Gly Val Val Val Asp Gly Leu Ala Gly Val Trp Pro Pro Arg
1105 1110 1115 1120
Gly Ala Val Ala Val Asp Val Asp Gly Val Arg Asp Arg Leu Ala Gly
1125 1130 1135
Ala Gly Cys Val Leu Gly Pro Val Phe Ser Gly Leu Arg Ala Val Trp
1140 1145 1150
Arg Asp Gly Gly Asp Leu Leu Ala Glu Val Cys Leu Pro Glu Glu Ala
1155 1160 1165
Trp Gly Asp Ala Ala Gly Phe Gly Leu His Pro Ala Leu Leu Asp Gly
1170 1175 1180
Val Val Gln Pro Leu Ser Val Leu Leu Pro Gly Gly Thr Gly Phe Gly
1185 1190 1195 1200
Glu Gly Ala Gly Phe Gly Glu Gly Val Arg Val Pro Ala Val Trp Gly
1205 1210 1215
Gly Val Ser Leu His Arg Ala Gly Val Thr Gly Val Arg Val Arg Val
1220 1225 1230
Trp Ala Val Gly Arg Gly Gly Gly Arg Glu Ala Val Ser Val Val Val
1235 1240 1245
Gly Asp Glu Ala Gly Val Pro Val Ala Ser Val Asp Arg Leu Glu Leu
1250 1255 1260
Arg Pro Val Asp Met Gly Gln Leu Arg Ala Val Ser Val Ser Ala Gly
1265 1270 1275 1280
Arg Arg Gly Ser Leu Tyr Ala Val Gln Trp Ala Glu Val Gly Pro Val
1285 1290 1295
Pro Val Cys Gly Gln Ala Trp Ala Trp His Glu Asp Val Gly Glu Ser
1300 1305 1310
Gly Gly Gly Pro Val Pro Gly Val Val Val Leu Arg Cys Pro Asp Ala
1315 1320 1325
Gly Ala Gly Gly Gly Gly Gly Gly Gly Val Gly Glu Val Val Gly Gly
1330 1335 1340
Val Leu Gly Val Val Gln Gly Trp Leu Gly Leu Glu Arg Phe Ala Gly
1345 1350 1355 1360
Ser Arg Leu Val Val Val Thr Arg Gly Ala Val Val Ala Gly Gln Glu
1365 1370 1375
Asp Gly Pro Val Asp Val Val Gly Ala Ala Val Trp Gly Leu Val Arg
1380 1385 1390
Ser Ala Gln Ala Glu His Pro Asp Arg Phe Val Leu Leu Asp Leu Asp
1395 1400 1405
Thr Asp Thr Asp Thr Gly Thr Asp Leu Asp Thr Gly Ala Gly Ala Gly
1410 1415 1420
Ala Gly Ala Gly Trp Gly Val Asp Gly Gly His Val Ala Ala Val Val
1425 1430 1435 1440
Ala Cys Gly Glu Pro Gln Leu Ala Val Arg Gly Glu Arg Val Leu Ala
1445 1450 1455
Ala Arg Leu Thr Arg Leu Glu Ser Ser Val Asp Val Pro Ala Gln Arg
1460 1465 1470
Ser Gly Asp Val Ala Gly Arg Glu Val Leu Pro Trp Leu Ser Gly Gly
1475 1480 1485
Ser Val Leu Val Thr Gly Gly Thr Gly Val Leu Gly Ala Ala Val Ala
1490 1495 1500
Arg His Leu Ala Gly Val Cys Gly Val Arg Asp Leu Leu Leu Val Ser
1505 1510 1515 1520
Arg Arg Gly Pro Asp Ala Pro Gly Ala Glu Gly Leu Arg Ala Glu Leu
1525 1530 1535
Ala Ala Leu Gly Ala Glu Val Arg Ile Val Ala Cys Asp Val Gly Glu
1540 1545 1550
Arg Arg Glu Val Val Arg Leu Leu Glu Gly Val Pro Ala Gly Cys Pro
1555 1560 1565
Leu Thr Gly Val Val His Ala Ala Gly Val Leu Asp Asp Ala Thr Ile
1570 1575 1580
Ala Ser Leu Thr Pro Glu Arg Leu Gly Thr Val Phe Ala Ala Lys Val
1585 1590 1595 1600
Asp Ala Ala Leu Leu Leu Asp Glu Leu Thr Arg Gly Met Glu Leu Ser
1605 1610 1615
Ala Phe Val Leu Phe Ser Ser Ala Ala Gly Ile Leu Gly Ser Ala Gly
1620 1625 1630
Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu Ala Tyr
1635 1640 1645
Arg Arg Arg Ala Ala Gly Leu Pro Gly Val Ser Leu Ala Trp Gly Leu
1650 1655 1660
Trp Glu Glu Ala Ser Gly Met Thr Gly His Leu Ala Gly Thr Asp His
1665 1670 1675 1680
Arg Arg Ile Ile Arg Ser Gly Leu His Pro Met Ser Thr Pro Asp Ala
1685 1690 1695
Leu Ala Leu Phe Asp Ala Ala Leu Ala Leu Asp Arg Pro Val Leu Leu
1700 1705 1710
Pro Ala Asp Leu Arg Pro Ala Pro Pro Leu Pro Pro Leu Leu Gln Asp
1715 1720 1725
Leu Leu Pro Ala Thr Arg Arg Arg Thr Thr Arg Thr Thr Thr Thr Gly
1730 1735 1740
Gly Ala Asp Asn Gly Ala Gln Leu His Ala Arg Leu Ala Gly Gln Thr
1745 1750 1755 1760
His Glu Gln Gln His Thr Thr Leu Leu Ala Leu Val Arg Ser His Ile
1765 1770 1775
Ala Thr Val Leu Gly His Thr Thr Pro Asp Thr Ile Pro Pro Asp Arg
1780 1785 1790
Ala Phe Arg Asp Leu Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Arg
1795 1800 1805
Asn Arg Leu Ser Arg Thr Thr Gly Leu Arg Leu Pro Thr Thr Leu Ala
1810 1815 1820
Phe Asp His Pro Asn Pro Thr Thr Leu Thr His His Leu His Thr Gln
1825 1830 1835 1840
Leu Leu Gly Ser Asp Ser Thr Ala Ser Ile Pro Ala Pro Arg Ala Ala
1845 1850 1855
Ala Val Pro Ala Asp Gln Asp Glu Pro Val Ala Ile Ile Gly Met Ala
1860 1865 1870
Cys Arg Tyr Pro Gly Gly Val Thr Ser Ala Glu Glu Leu Trp Glu Leu
1875 1880 1885
Leu Ala Ser Gly Arg Asp Thr Val Gly Glu Phe Pro Thr Asp Arg Gly
1890 1895 1900
Trp Asp Leu Glu Ala Leu Phe Asp Pro Glu Pro Gly Arg Pro Gly Thr
1905 1910 1915 1920
Ser Tyr Thr Arg Cys Gly Ser Phe Leu Tyr Asp Ala Gly Glu Phe Asp
1925 1930 1935
Ala Gly Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro
1940 1945 1950
Gln Gln Arg Leu Leu Leu Glu Ala Ser Trp Glu Ala Met Glu Gln Ala
1955 1960 1965
Gly Ile Asp Pro Thr Thr Val Arg Gly Ser Gln Thr Gly Val Phe Ala
1970 1975 1980
Gly Leu Ile Pro Gln Ala Tyr Gly Pro Arg Leu His Glu Asn Ala Ala
1985 1990 1995 2000
Ala Asp Thr Glu Gly Tyr Val Leu Thr Gly Thr Ser Gly Ser Val Ala
2005 2010 2015
Ser Gly Arg Ile Ser Tyr Thr Phe Gly Phe Glu Gly Pro Ala Val Ser
2020 2025 2030
Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys
2035 2040 2045
Gln Ala Leu Arg Ala Gly Glu Cys Ser Met Ala Leu Ala Gly Gly Val
2050 2055 2060
Thr Val Met Ser Ser Pro Gly Ala Phe Val Glu Phe Ser Arg Gln Arg
2065 2070 2075 2080
Gly Leu Ala Ala Asp Gly His Cys Lys Ala Phe Ser Ala Ala Ala Asp
2085 2090 2095
Gly Thr Gly Trp Gly Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu
2100 2105 2110
Ser Asp Ala Arg Arg Asn Gly His Arg Val Leu Ala Val Val Arg Gly
2115 2120 2125
Ser Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn
2130 2135 2140
Gly Pro Ser Gln Gln Arg Val Ile Arg Gln Ala Leu Ala Asn Ala Gly
2145 2150 2155 2160
Leu Ser Ala Gly Asp Val Asp Ala Val Glu Ala His Gly Thr Gly Thr
2165 2170 2175
Thr Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly
2180 2185 2190
Gln Asp Arg Ala Gly Glu Gly Pro Leu Trp Leu Gly Ser Val Lys Ser
2195 2200 2205
Asn Val Gly His Thr Gln Ala Ala Ala Gly Val Ala Gly Val Ile Lys
2210 2215 2220
Met Val Met Ala Leu Arg Asn Gly Leu Leu Pro Arg Thr Leu His Val
2225 2230 2235 2240
Asp Glu Pro Ser Pro His Val Asp Trp Ser Ala Gly Ala Val Gln Leu
2245 2250 2255
Leu Thr Glu Thr Val Pro Trp Pro Gly Gly Glu Gly Arg Leu Arg Arg
2260 2265 2270
Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val Ile
2275 2280 2285
Leu Glu Glu Ala Pro Ala His Asn Ile Pro Ser Asp Thr Pro Ala Asp
2290 2295 2300
Asp Val Pro Gly Gly Pro Pro Ala Gly Glu Asp Ala Gly Ser Gly Glu
2305 2310 2315 2320
Glu Ala Ala Ala Gly Ser Pro Gly Val Trp Pro Trp Leu Val Ser Ala
2325 2330 2335
Lys Ser Gln Pro Ala Leu Arg Ala Gln Ala Gln Ala Leu His Ala His
2340 2345 2350
Leu Thr Asp His Pro Gly Leu Asp Leu Ala Asp Val Gly Tyr Thr Leu
2355 2360 2365
Ala His Ala Arg Ala Val Phe Asp His Arg Ala Thr Leu Ile Ala Ala
2370 2375 2380
Asp Arg Asp Thr Phe Leu Gln Ala Leu Gln Ala Leu Ala Ala Gly Glu
2385 2390 2395 2400
Pro His Pro Ala Val Ile His Ser Ser Ala Pro Gly Gly Thr Gly Thr
2405 2410 2415
Gly Glu Ala Ala Gly Lys Thr Ala Phe Ile Cys Ser Gly Gln Gly Thr
2420 2425 2430
Gln Arg Pro Gly Met Ala His Gly Leu Tyr His Thr His Pro Val Phe
2435 2440 2445
Ala Ala Ala Leu Asn Asp Ile Cys Thr His Leu Asp Pro His Leu Asp
2450 2455 2460
His Pro Leu Leu Pro Leu Leu Thr Gln Asp Pro Asn Thr Gln Asp Thr
2465 2470 2475 2480
Thr Thr Leu Glu Glu Ala Ala Ala Leu Leu Gln Gln Thr Pro Tyr Ala
2485 2490 2495
Gln Pro Ala Leu Phe Ala Phe Gln Val Ala Leu His Arg Leu Leu Thr
2500 2505 2510
Asp Gly Tyr His Ile Thr Pro His Tyr Tyr Ala Gly His Ser Leu Gly
2515 2520 2525
Glu Ile Thr Ala Ala His Leu Ala Gly Ile Leu Thr Leu Thr Asp Ala
2530 2535 2540
Thr Thr Leu Ile Thr Gln Arg Ala Thr Leu Met Gln Thr Met Pro Pro
2545 2550 2555 2560
Gly Thr Met Thr Thr Leu His Thr Thr Pro His His Ile Thr His His
2565 2570 2575
Ile Thr Ala His Glu Asn Asp Leu Ala Ile Ala Ala Ile Asn Thr Pro
2580 2585 2590
Thr Ser Leu Val Ile Ser Gly Thr Pro His Thr Val Gln His Ile Thr
2595 2600 2605
Thr Leu Cys Gln Gln Gln Gly Ile Lys Thr Lys Thr Leu Pro Thr Asn
2610 2615 2620
His Ala Phe His Ser Pro His Thr Asn Pro Ile Leu Asn Gln Leu His
2625 2630 2635 2640
Gln His Thr Gln Thr Leu Thr Tyr His Pro Pro His Thr Pro Leu Ile
2645 2650 2655
Thr Ala Asn Thr Pro Pro Asp Gln Leu Leu Thr Pro His Tyr Trp Thr
2660 2665 2670
Gln Gln Ala Arg Asn Thr Val Asp Ile Ala Thr Thr Thr Gln Thr Leu
2675 2680 2685
His Gln His Gly Val Thr Thr Tyr Ile Glu Leu Gly Pro Asp Asn Thr
2690 2695 2700
Leu Thr Thr Leu Thr His His Asn Leu Pro Asn Thr Pro Thr Thr Thr
2705 2710 2715 2720
Leu Thr Leu Thr His Pro His His His Pro Gln Thr His Leu Leu Thr
2725 2730 2735
Asn Leu Ala Lys Thr Thr Thr Thr Trp His Pro His His Tyr Thr His
2740 2745 2750
His His Asn Gln Pro His Thr His Thr His Leu Asp Leu Pro Thr Tyr
2755 2760 2765
Pro Phe Gln His His His Tyr Trp Leu Glu Ser Thr Gln Pro Gly Ala
2770 2775 2780
Gly Asn Val Ser Ala Ala Gly Leu Asp Pro Thr Glu His Pro Leu Leu
2785 2790 2795 2800
Gly Ala Thr Leu Glu Leu Ala Glu Gly Asp Gly Cys Leu Leu Thr Gly
2805 2810 2815
Arg Leu Ser Leu Arg Thr His Pro Trp Leu Ala Gly His Ala Val Gly
2820 2825 2830
Gly Val Val Leu Leu Pro Gly Thr Ala Phe Ala Glu Leu Ala Leu His
2835 2840 2845
Ala Gly Glu Ser Val Gly Cys Asp His Val Asp Glu Leu Thr Leu His
2850 2855 2860
Thr Pro Leu Val Ile Pro Glu Val Gly Asp Val Thr Leu Gln Val Ala
2865 2870 2875 2880
Ile Ala Ala Pro Asp Glu Ser Gly Arg Arg Met Met Thr Ile His Ser
2885 2890 2895
Arg Gly Glu Gly Gly Ser Gly Gly Ala Asp Ala Ser Ala Ser Ala Trp
2900 2905 2910
Thr Arg His Ala Ala Gly Val Leu Ser Pro Ala Lys Asp Asp Asp Thr
2915 2920 2925
Ala Ser Tyr Glu Leu Leu Ala Gly Pro Trp Pro Pro Val Gly Ala Thr
2930 2935 2940
Pro Val Asp Leu Asn Thr Ala Tyr Asp Gln Met Ala Asp Ala Gly Phe
2945 2950 2955 2960
Ala Tyr Gly Leu Ala Phe Gln Gly Leu Arg Ala Ala Trp Arg Tyr Gly
2965 2970 2975
Asp Asp Ile Leu Val Glu Ala Arg Leu Pro Glu Glu Val Ser Gly Asp
2980 2985 2990
Ala Ala Ala Tyr Gly Leu His Pro Ala Leu Leu Asp Ala Ala Leu Gln
2995 3000 3005
Gly Thr Gly Leu Leu Ser Val Ala Gly Pro Gly Thr Pro Val Val Pro
3010 3015 3020
His Val Trp Asn Gly Leu Arg Phe Arg Thr His Gly Ala Val Ser Val
3025 3030 3035 3040
Arg Ala Cys Leu Ser Thr Leu Gly Ala Thr Gly Ala Ala Val Cys Val
3045 3050 3055
Arg Ile Thr Asp Asp Thr Gly Val Pro Val Ala Ser Val Asp Arg Leu
3060 3065 3070
Glu Leu Arg Pro Val Asp Met Gly Gln Leu Arg Ala Val Ser Val Ser
3075 3080 3085
Ala Gly Arg Arg Gly Ser Leu Tyr Ala Val Gln Trp Ala Glu Val Gly
3090 3095 3100
Pro Val Pro Val Cys Gly Gln Ala Trp Ala Trp His Glu Asp Val Gly
3105 3110 3115 3120
Glu Ser Gly Gly Gly Pro Val Pro Gly Val Val Val Leu Arg Cys Pro
3125 3130 3135
Asp Ala Gly Ala Asp Gly Gly Gly Gly Gly Gly Val Gly Glu Val Val
3140 3145 3150
Gly Gly Val Leu Gly Val Val Gln Gly Trp Leu Gly Leu Glu Arg Phe
3155 3160 3165
Ala Gly Ser Arg Leu Val Val Val Thr Arg Gly Ala Val Val Ala Gly
3170 3175 3180
Pro Glu Asp Gly Pro Val Asp Val Val Gly Ala Ala Val Trp Gly Leu
3185 3190 3195 3200
Val Arg Ser Ala Gln Ala Glu His Pro Asp Arg Phe Val Leu Leu Asp
3205 3210 3215
Leu Asp Thr Asp Leu Asp Ser Gly Ala Asp Ala Asp Ala Gly Asn Glu
3220 3225 3230
Ala Gly Met Gly Ser Gly Leu Asp Gly Gly Arg Val Ala Ala Val Val
3235 3240 3245
Ala Cys Gly Glu Pro Gln Leu Ala Val Arg Gly Glu Arg Val Leu Ala
3250 3255 3260
Ala Arg Leu Thr Arg Leu Glu Ser Pro Val Asp Val Ser Gly Arg Glu
3265 3270 3275 3280
Val Leu Pro Trp Leu Ser Gly Gly Ser Val Leu Val Thr Gly Gly Thr
3285 3290 3295
Gly Val Leu Gly Ala Ala Val Ala Arg His Leu Ala Gly Val Cys Gly
3300 3305 3310
Val Arg Asp Leu Leu Leu Val Ser Arg Arg Gly Pro Asp Ala Pro Gly
3315 3320 3325
Ala Glu Gly Leu Arg Ala Glu Leu Ala Ala Leu Gly Ala Glu Val Arg
3330 3335 3340
Ile Val Ala Cys Asp Val Gly Glu Arg Arg Glu Val Val Arg Leu Leu
3345 3350 3355 3360
Glu Gly Val Pro Ala Gly Cys Pro Leu Thr Gly Val Val His Ala Ala
3365 3370 3375
Gly Val Leu Asp Asp Ala Thr Ile Ala Ser Leu Thr Pro Glu Arg Leu
3380 3385 3390
Gly Thr Val Phe Ala Ala Lys Val Asp Ala Ala Leu Leu Leu Asp Glu
3395 3400 3405
Leu Thr Arg Gly Met Glu Leu Ser Ala Phe Val Leu Phe Ser Ser Ala
3410 3415 3420
Ala Gly Ile Leu Gly Ser Ala Gly Gln Gly Asn Tyr Ala Ala Ala Asn
3425 3430 3435 3440
Ala Ala Leu Asp Ala Leu Ala Tyr Arg Arg Arg Ala Ala Gly Leu Pro
3445 3450 3455
Gly Val Ser Leu Ala Trp Gly Leu Trp Glu Glu Ala Ser Gly Met Thr
3460 3465 3470
Gly His Leu Ala Gly Thr Asp His Arg Arg Ile Ile Arg Ser Gly Leu
3475 3480 3485
His Pro Met Ser Thr Pro Asp Ala Leu Ala Leu Phe Asp Ala Ala Leu
3490 3495 3500
Ala Leu Asp Arg Pro Val Leu Leu Pro Ala Asp Leu Arg Pro Ala Pro
3505 3510 3515 3520
Pro Leu Pro Pro Leu Leu Gln Asp Leu Leu Pro Ala Thr Arg Arg Arg
3525 3530 3535
Thr Thr Arg Thr Thr Thr Thr Gly Gly Ala Asp Asn Gly Ala Gln Leu
3540 3545 3550
His Ala Arg Leu Ala Gly Gln Thr His Glu Gln Gln His Thr Thr Leu
3555 3560 3565
Leu Ala Leu Val Arg Ser His Ile Ala Thr Val Leu Gly His Asn Ala
3570 3575 3580
Pro Glu Met Ile Pro Val Asp Ser Ala Phe Arg Asp Leu Gly Phe Asp
3585 3590 3595 3600
Ser Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Gly Glu Ala Thr Gly
3605 3610 3615
Leu Arg Leu Pro Thr Ser Leu Val Phe Asp Gln Pro Asn Ala Ala Thr
3620 3625 3630
Leu Ala Arg His Leu Arg Arg Glu Leu Met Gly Asp Asp Ala Glu Gly
3635 3640 3645
Glu Thr Pro Ser Gln Val Ala Leu His Gln Val Ala Ala Asp Glu Pro
3650 3655 3660
Ile Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly Val Cys Ser
3665 3670 3675 3680
Pro Glu Glu Leu Trp Glu Leu Val Ala Ser Gly Gly Asp Ala Ile Gly
3685 3690 3695
Glu Phe Pro Ala Gly Arg Gly Trp Asp Leu Glu Gly Leu Phe Asp Ser
3700 3705 3710
Asp Pro Asp Arg Ser Gly Thr Ser Tyr Ala Arg Tyr Gly Gly Phe Leu
3715 3720 3725
Tyr Glu Ala Gly Glu Phe Asp Ala Asp Phe Phe Gly Ile Ser Pro Arg
3730 3735 3740
Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Thr Ser
3745 3750 3755 3760
Trp Glu Ala Phe Glu Arg Ala Gly Ile Asp Pro Leu Ser Met Arg Gly
3765 3770 3775
Ser Arg Thr Gly Val Phe Ala Gly Val Met Tyr His Asp Tyr Ala Ala
3780 3785 3790
Arg Leu His His Val Pro Glu Gly Phe Glu Gly Leu Ile Ala Asn Gly
3795 3800 3805
Ser Ala Gly Ser Val Ala Thr Gly Arg Val Ala Tyr Ser Phe Gly Leu
3810 3815 3820
Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val
3825 3830 3835 3840
Ala Leu His Trp Ala Ala Gln Ala Leu Arg Ala Gly Glu Cys Ser Met
3845 3850 3855
Ala Leu Ala Gly Gly Val Thr Val Met Ser Ser Pro Gly Thr Phe Val
3860 3865 3870
Glu Phe Ser Arg Gln Arg Gly Leu Ala Ala Asp Gly Arg Cys Lys Ala
3875 3880 3885
Tyr Ser Ala Ala Ala Asp Gly Thr Gly Trp Ala Glu Gly Val Gly Met
3890 3895 3900
Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Asn Gly His Arg Val
3905 3910 3915 3920
Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn
3925 3930 3935
Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln Arg Val Ile Arg Gln
3940 3945 3950
Ala Leu Ala Asn Ala Gly Leu Thr Pro Ala Asp Val Asp Ala Val Glu
3955 3960 3965
Gly His Gly Thr Gly Thr Thr Leu Gly Asp Pro Ile Glu Ala Gln Ala
3970 3975 3980
Leu Leu Ala Ala Tyr Gly Gln His Arg Pro His His Arg Pro Leu Trp
3985 3990 3995 4000
Leu Gly Ser Leu Lys Ser Asn Ile Gly His Ala Gln Ala Ala Ala Gly
4005 4010 4015
Val Gly Gly Val Ile Lys Met Val Met Ala Leu Arg Asn Gly Leu Leu
4020 4025 4030
Pro Gln Thr Leu His Val Asp Glu Pro Thr Pro Gln Val Asp Trp Ser
4035 4040 4045
Thr Gly Ala Val Gln Leu Leu Thr Gln Pro Val Pro Trp Pro Ala Asp
4050 4055 4060
Pro Ala Gly Arg Pro Arg His Ala Gly Val Ser Ser Phe Gly Val Ser
4065 4070 4075 4080
Gly Thr Asn Ala His Val Ile Leu Glu Glu Ala Pro Ala Ala Ala Gly
4085 4090 4095
Gly Ala Ala Gly Gly Gly Val Ser Val Gly Ala Pro Asn Pro Ala Leu
4100 4105 4110
Pro Val Ala Glu Ser Glu Pro Val Pro Val Pro Val Pro Val Ser Ala
4115 4120 4125
Arg Ser Glu Ala Gly Leu Arg Ala Gln Ala Gln Ala Leu Arg Gln Tyr
4130 4135 4140
Val Ala Ala Arg Pro Asp Met Ser Pro Ala Asp Ile Gly Ala Gly Leu
4145 4150 4155 4160
Ala Arg Gly Arg Ala Val Leu Glu His Arg Ala Val Ile Leu Ala Ala
4165 4170 4175
Asp Arg Glu Glu Leu Ala Gln Ala Leu Thr Ala Leu Ala Ala Gly Glu
4180 4185 4190
Pro His Pro His Ile Thr Thr Gly His Thr Arg Gly Ser Asp Arg Gly
4195 4200 4205
Gly Val Val Phe Val Phe Pro Gly Gln Gly Gly Gln Trp Ala Gly Met
4210 4215 4220
Gly Leu Thr Leu Leu Thr Ser Ser Pro Val Phe Ala Glu His Ile Asp
4225 4230 4235 4240
Ala Cys Glu Lys Ala Leu Thr Pro Trp Val Pro Trp Ser Leu Thr Asp
4245 4250 4255
Ile Leu His Arg Asp Pro Asp Asp Pro Ala Trp Gln Gln Ala Asp Val
4260 4265 4270
Val Gln Pro Val Leu Phe Ser Ile Met Val Ser Leu Ala Ala Leu Trp
4275 4280 4285
Arg Ser Tyr Gly Ile Glu Pro Asp Ala Val Leu Gly His Ser Gln Gly
4290 4295 4300
Glu Ile Ala Ala Ala His Ile Cys Gly Ala Leu Ser Leu Lys Asp Ala
4305 4310 4315 4320
Ala Lys Thr Val Ala Leu Arg Ser Gln Ala Leu Ala Ala Val Arg Gly
4325 4330 4335
Arg Gly Ala Met Val Ser Leu Pro Leu Pro Ala Gln Asp Val Gln Gln
4340 4345 4350
Leu Ile Ser Glu Arg Trp Glu Gly Gln Leu Trp Val Ala Ala Leu Asn
4355 4360 4365
Gly Pro His Ser Thr Thr Val Ser Gly Asp Thr Thr Ala Val Glu Glu
4370 4375 4380
Leu Leu Thr His Cys Ala Asp Thr Gly Leu Arg Ala Lys Arg Ile Pro
4385 4390 4395 4400
Val Asp Tyr Ala Ser His Cys Pro His Val Gln Pro Leu His Asp Glu
4405 4410 4415
Leu Leu His Leu Leu Gly Asp Ile Thr Pro Gln Pro Ser Thr Met Pro
4420 4425 4430
Phe Phe Ser Thr Val Val Gly His Leu Val Trp Tyr Thr Thr Thr Leu
4435 4440 4445
Asp Ala Ala Tyr Trp Tyr Arg Asn Leu His Gln Pro Val Arg Phe Ser
4450 4455 4460
His Ala Ile Gln Thr Leu Thr Asp Asp Gly His Arg Pro Phe Ile Glu
4465 4470 4475 4480
Ile Ser Pro His Pro Thr Leu Val Pro Ala Ile Glu Asp Thr Thr Glu
4485 4490 4495
Asn Thr Thr Glu Asn Ile Thr Ala Thr Gly Ser Leu Arg Arg Gly Asp
4500 4505 4510
Asn Asp Thr His Arg Phe Leu Thr Ala Leu Ala His Thr His Thr Thr
4515 4520 4525
Gly Ile Arg Thr Pro Thr Thr Trp His His His Tyr Thr Gln Thr His
4530 4535 4540
Pro His Pro His Asn His His Leu Asp Leu Pro Thr Tyr Pro Phe Gln
4545 4550 4555 4560
His Gln His Tyr Trp Leu Gln Pro Pro Thr Thr Thr Thr Asp Leu Thr
4565 4570 4575
Thr Thr Gly Leu Thr Pro Thr His His Pro Leu Leu Thr Ala Thr Leu
4580 4585 4590
Thr Leu Ala Asn Asn Asn Thr Gln Leu Leu Thr Gly Arg Leu Ser Leu
4595 4600 4605
Arg Thr His Pro Trp Leu Thr Asp His Thr Val Val Gly Thr Thr Leu
4610 4615 4620
Val Pro Gly Thr Ala Leu Leu Glu Leu Ala Leu Gln Ala Thr Thr Thr
4625 4630 4635 4640
Asp His Leu Glu Glu Leu Ala Leu His Thr Pro Leu Val Ile Pro Arg
4645 4650 4655
Glu Gly Ala Val Asp Val Gln Val His Ile Asn Pro Pro Asp Asp Thr
4660 4665 4670
Asp Thr Arg Ser Leu Thr Ile Tyr Ser Arg Ser Glu Asn Ala Pro Ala
4675 4680 4685
Ala Ala Pro Trp Arg His His Ala Thr Ala Val Leu Gly Thr Lys Thr
4690 4695 4700
Ser Arg Ile Glu Thr Gly Arg Ser His Asp Asp Leu Ser Met Trp Pro
4705 4710 4715 4720
Pro Ala Gly Ala Val Arg Cys Ala Asp Glu Glu Leu Ala Ala Leu Tyr
4725 4730 4735
Gly Asp Tyr Glu Ala Asn Gly Phe Val Tyr Gly Pro Ala Phe Arg Gly
4740 4745 4750
Leu Thr Ala Ala Trp Arg Leu Gly Asp Glu Val Phe Ala Glu Val Arg
4755 4760 4765
Leu Pro Glu Gln Val His Gly Glu Ala Ser Ala Tyr Asn Leu His Pro
4770 4775 4780
Ala Leu Leu Asp Ala Ala Leu His Ala Ala Ala Phe Ala Pro Ser Gly
4785 4790 4795 4800
Ser Leu Pro Gln Gly Ser Val Pro Phe Ser Phe Thr Gly Val Thr Leu
4805 4810 4815
His Ala Ala Asn Ala Ser Ser Leu Arg Val Arg Leu Ser Pro Ala Asp
4820 4825 4830
Pro Asn Ser Gly His Ala Ala Val Ser Val Leu Val Thr Asp Asp Thr
4835 4840 4845
Gly Thr Pro Val Ala Ser Val Glu Ala Leu Ala Val Arg Pro Leu Ala
4850 4855 4860
Ala Asp Glu Leu Arg Ala Ala Glu Arg Ala Val Gln Arg Ala Glu Leu
4865 4870 4875 4880
Phe Asp Met Lys Trp Val Glu Val Pro Ser Asp Val Leu Val Ser Gly
4885 4890 4895
Gly Ala Ser Val Val Val Leu Asp Gly Ala Asp Asp Leu Val Gly Leu
4900 4905 4910
Ala Ala Glu Glu Asp Gly Val Pro Gly Val Val Val Leu Arg Cys Pro
4915 4920 4925
Asp Ala Gly Ala Asp Gly Gly Gly Gly Gly Gly Gly Val Gly Glu Val
4930 4935 4940
Val Gly Gly Val Leu Gly Val Val Gln Gly Trp Leu Gly Leu Glu Arg
4945 4950 4955 4960
Phe Ala Gly Ser Arg Leu Val Val Val Thr Arg Gly Ala Val Val Ala
4965 4970 4975
Gly Pro Glu Asp Gly Pro Val Asp Gly Pro Val Asp Val Val Gly Ala
4980 4985 4990
Ala Val Trp Gly Leu Val Arg Ser Ala Gln Ala Glu His Pro Asp Arg
4995 5000 5005
Phe Val Leu Leu Asp Leu Asp Thr Asp Leu Asp Ser Gly Ala Asp Arg
5010 5015 5020
Asp Ala Gly Asn Glu Ala Gly Met Gly Ser Gly Leu Asp Gly Gly Arg
5025 5030 5035 5040
Val Ala Ala Val Val Ala Cys Gly Glu Pro Gln Leu Ala Val Arg Gly
5045 5050 5055
Glu Arg Val Leu Ala Ala Arg Leu Thr Arg Leu Glu Ser Pro Val Asp
5060 5065 5070
Val Ser Gly Arg Glu Val Leu Pro Trp Leu Ser Gly Gly Ser Val Leu
5075 5080 5085
Val Thr Gly Gly Thr Gly Val Leu Gly Ala Ala Val Ala Arg His Leu
5090 5095 5100
Ala Gly Val Cys Gly Val Arg Asp Leu Leu Leu Val Ser Arg Arg Gly
5105 5110 5115 5120
Pro Asp Ala Pro Gly Ala Glu Gly Leu Arg Ala Glu Leu Ala Ala Leu
5125 5130 5135
Gly Ala Glu Val Arg Ile Val Ala Cys Asp Val Gly Glu Arg Arg Glu
5140 5145 5150
Val Val Arg Leu Leu Glu Gly Val Pro Ala Gly Cys Pro Leu Thr Gly
5155 5160 5165
Val Val His Ala Ala Gly Val Leu Asp Asp Ala Thr Ile Ala Ser Leu
5170 5175 5180
Thr Pro Glu Arg Leu Gly Thr Val Phe Ala Ala Lys Val Asp Ala Ala
5185 5190 5195 5200
Leu Leu Leu Asp Glu Leu Thr Arg Gly Met Glu Leu Ser Ala Phe Val
5205 5210 5215
Leu Phe Ser Ser Ala Ala Gly Ile Leu Gly Ser Ala Gly Gln Gly Asn
5220 5225 5230
Tyr Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu Ala Tyr Arg Arg Arg
5235 5240 5245
Ala Ala Gly Leu Pro Gly Val Ser Leu Ala Trp Gly Leu Trp Glu Glu
5250 5255 5260
Ala Ser Gly Met Thr Gly His Leu Ala Gly Thr Asp His Arg Arg Ile
5265 5270 5275 5280
Ile Arg Ser Gly Leu His Pro Met Ser Thr Pro Asp Ala Leu Ala Leu
5285 5290 5295
Phe Asp Ala Ala Leu Ala Leu Asp Arg Pro Val Leu Leu Pro Ala Asp
5300 5305 5310
Leu Arg Pro Ala Pro Pro Leu Pro Pro Leu Leu Gln Asp Leu Leu Pro
5315 5320 5325
Ala Thr Arg Arg Arg Thr Thr Arg Thr Thr Thr Thr Gly Gly Ala Asp
5330 5335 5340
Asn Gly Ala Gln Leu His Gly Arg Leu Ala Gly Gln Thr His Glu Gln
5345 5350 5355 5360
Gln His Thr Thr Leu Leu Ala Leu Val Arg Ser His Ile Ala Thr Val
5365 5370 5375
Leu Gly His Thr Thr Pro Asp Thr Ile Pro Pro Asp Arg Ala Phe Arg
5380 5385 5390
Asp Leu Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Arg Asn Arg Leu
5395 5400 5405
Ser His Thr Thr Gly Leu Arg Leu Pro Thr Thr Leu Ala Phe Asp His
5410 5415 5420
Pro Asn Pro Thr Thr Leu Thr His His Leu His Thr Gln Leu Val Ser
5425 5430 5435 5440
Lys Gly Leu Thr Ala Ala Ala Glu Pro Asp Ala Ala Thr Thr Pro Pro
5445 5450 5455
Gly Leu Pro Ser Leu Leu Ser Glu Leu Glu Arg Leu Glu Ala Val Val
5460 5465 5470
Leu Ser Ser Thr Thr Ser Ser Ala Ala Pro Leu Asp Asp Gly Ala Arg
5475 5480 5485
Thr Arg Leu Ala Ser Arg Leu His Ser Leu Ala Gln Lys Leu Asn Gly
5490 5495 5500
Asp Asp Thr Ala Pro Asp Leu Ala Glu Thr Ser Asp Glu Glu Met Phe
5505 5510 5515 5520
Ala Leu Ile Asp Arg Glu Val Gly Phe Glu Ser Gln
5525 5530
<210> 73
<211> 17460
<212> DNA
<213> Artificial Sequence
<220>
<223> milA3 gene of Streptomyces milbemycinicus
<400> 73
atggccgctg gccacgacaa ggtgatcgag gcgctgcggg cgtccctcaa gaccaacgag 60
cggcagaggg aacagatcca ccggctcact acggcggcgc gggaacccat cgccatcatc 120
ggcatggcct gccgctatcc gggcggagtg ggatcgccgg aggacctgtg ggagctggtg 180
gccgccggtc gtgacgccat cggcaccttc cccgaggacc ggggctggga cgtggagcgg 240
ctgtacgacc ccgatccgga gcgggccggc acctcgtgta cccagcatgg cggattcctg 300
taccaggcag gggagttcga ccccggtttc ttcgggatca gcccgcgcga ggcgctggcg 360
atggacccgc agcagcggct gctgctggag atctcctggg aggtgttcga gcgggccggg 420
atcgacccgg cctcggtgcg cggcagccgc accggggtgt tcgcgggcgt catgtaccac 480
gactacggct cccggctgca caccgtcccc gaaggcttcg agggctatgt cggcaacggc 540
agcggcggcg gcgtggcgtc cggccgggtc gcctacaccc tcggcctcga aggcccggcc 600
gtgaccgtgg acaccgcctg ctcctcctcg ttggtcgccc tgcacctggc ctgccaggcg 660
ctgcgggccg gcgagtgctc actcgccctg gcgggcgggg tgacggtgat gtccaccccc 720
agcctgttcg tcgagtactc ccggcagcgc gcgctcgcgg cagacggccg gtgcaaggcg 780
tacggggcgg gggcggacgg caccggctgg gcagaaggcg ccgggatgct gctggtggaa 840
cggctcacgg acgcacagcg cctcggccac cgggtgctgg cggtggtccg gggcagcgcg 900
gtcaaccagg acggcgcgag caacggcctc accgccccca acggccccgc gcaacaacgg 960
gccatccggc aggcactggc gagcgccggg gtgtcggcgt ccgaggtcga cgccgtggag 1020
gggcatggga cggggacgcg gctgggcgat ccgatcgagg cgcaggcgtt gctggcgacc 1080
tacggtcagc agcggcccgc ggaccggccg ctgtggctcg ggtcgatgaa gtccaacgtc 1140
ggccatgcgc aggcggccgc cggcgtgggc gggatcatca agatggtgat ggccatgcgg 1200
agcgggacgc tgccgcgcac cctgcacgcg gacgagccgt cgccacacat cgactgggac 1260
tcgggcgcgg tacggctgct gaccgagccg gtcgcctggc cggagcgcga ccggccccgc 1320
cgcgccgcgg tgtcctcctt cggggtcagc ggcaccaacg cccatgtgat cctcgaggcc 1380
gcatcgcaga cggcgccgca gacggattcc gcgtcgcagg cggaaaccga cgacgctccc 1440
gcaccgcacg gcgcgccggg ccatgccgtg gcggggccgc tgctctggcc cttgtcgggc 1500
gcgacggccg aggcgctgcg ggcccaggcc ggggagctgc gtcgcttcgt ggcggccgat 1560
gagctgctgc gccccgccga cgtcgggcac accctggtct tcggccgctc ggacctcgca 1620
caccgcgcag tcgtcctcgg ctccgaccgg gaaaccctgc tgcgcgctct ggacactctg 1680
gcaggggagg ggccggacga cggctcggtc gtacggggca tggcggccgc cggggccggt 1740
gcgggcgtgg tgttcgtctt cccgggacag ggcggccagt gggccggcat ggggctgcgg 1800
ctgctggaga cctcgtcgtt cttcgccgag cggatggcgg agtgcgaggc ggcgttggca 1860
ccgtatgccg actggtcgct gctcgacgtt ctgcgccggg accccgggga cccggtctgg 1920
gagcgggccg atgtcgtcca gccgatgctg ttctcggtga tggtgtcgct ggcgcagctg 1980
tggcgctcgt acggcgtcga accggacgcc gtactcggcc actcccaggg cgagatcgcc 2040
gccgcccaca tctgcggcgc gctgaccctg gacgacgccg cgaaggttgt cgcgctgcgc 2100
agccgggccc tgcagaccct gcgcggttcg ggcggcatgg cctccgtacc actgccggcg 2160
gacgaggtca ccgggctgct gcggaccgca tggccggacc ggctgtgggt ggccgccgtc 2220
aacgccccca cggccacggt gatctccggc gacgcggact ctctggcgga ggcgctggaa 2280
cactaccggg accagggcgt cgaagcgaag cgggtcccgg tcgactacgc ctcccactgc 2340
ccgcatatcg aagccgtgga gcaggagctg ctgggcctgt tgcgggggat cgctccaagg 2400
gccgccgaca tccccttcta ctccaccgtg gacaaccagt gggccgacac catgggactc 2460
gacgcccggt actggtaccg caatctgcgc cggcccgtac gcttcgccga agcgctccgc 2520
gccctcggcg ccgccgagta ccggacgtat gtcgaggtcg gcccgcaccc caccctcacc 2580
cccgccatcg aggacaccac tgaggccgcc ggcgtcgcgg ccacggttgt cggatccctg 2640
cgccgcggcg aggacgacgc ccaccgcatc ctgacctcgc tggcccgggc tcatattcat 2700
ggcctgcccg tggcgtggga ccgccactac cgggcgctcg cccccgaggc gaaccatgtc 2760
gacctgccca cctacgcctt ccagcgccgc cgctactggc tggacgcccc ggcgaccacc 2820
ggggacgtga cggccgcggg gctggccccg gtcggacacc cactgctcgg cgcggcggtc 2880
ggactcgccg agggcgacgg atatctgctc accggccggc tcgccccgca cacccacccc 2940
tggctcaccg accacgcggt cgccggcacc gtcctgctgc cgggcaccgc atacgtggaa 3000
ctggccgtgc acgtcggcgg acacctcggc tgcccccggc tggaggagct caccctgcac 3060
gccccgctcg tcctccccga caccggcggc gtggcgctcc aggtggccgt cggggcaccg 3120
gacgagaccg gccgccgcgc actgagcgtc tacgcacagc gcgacgacga ccccgcgtgg 3180
gagggggcgg cccggggcgc gtggacacgg catgcgaccg gcacactggc ggccgaggcc 3240
ccgactgatg gcatcagcgg tgccgacggt gccgggaccc tggcgggggc gtggcctccg 3300
ccgggcgcgg agcccctgga catcagcggc ctctacgaca cgctggccgc cgcagacttc 3360
ggctacggcc cggccttcca ggggctgcgc gccgtctggc ggcaaggcga ggagacctac 3420
gccgaggtgc ggctccccga ccaggtggcc gccgacgccc cacgcttctg cctccacccc 3480
gcgctgctcg acgccgcgct ccacccgctg gcactcgaca gcggccgaag cgaggagaat 3540
ccagcgggac atggcctgct gccgttcgcc tggcgcggcg tcagcctgcg ctccccgggc 3600
acaccgacgc tgcgcgtacg gctgcggccg cagggcccgg actcgattgc cgtcgacgtg 3660
gccgacgaga cgggcgcgcc ggtggcctcg gccgaatcgc tcacgctgcg gccggtggcc 3720
ctggaggacc tgcgggccct cggcggccag gcgggcgaca ccctctacgc cctggagtgg 3780
accgccgcgc ccgagccccc ggcgacggcc ctcgggcggt gcgctgtgat tggccaagcc 3840
attcctggat gggctgccgc gctggagacg gcggcagcgg ggcccgtacg gcggtacccg 3900
gaccttgccg gactggtgac ggccctggac gcgggcgatc cgcctccgga cctggtgttc 3960
gtgggctgcc ctccggctgc cgccgggccc gacgacacga cggtcgccga cgtccacacc 4020
gcccgtaccc gtgtccgtac ccgacaagcg ctggacctgc ttcagggctg gctcggcgaa 4080
gcgcggctgg ccggcgcgag gctggtgctg gtcacctgcg gcgcggtggc caccgggccg 4140
gcggagggag tgatggacct ggcgggcgcg gcgatctgcg gactggtgcg atccgcgcag 4200
gccgaggagc ccgaccgtat cctcctggtg gacctggacg cggccgagga gtcgtgggcg 4260
gcgctaccac gggcggtcgc gctgggcgaa ccgcagatgg ccatccgggc cggccagccg 4320
cacatggccc ggctggttcg agccgacacc gaggggggcg ccctgctcac gccgccacag 4380
gggagcggcg gctggcggct cgactgcgcc gacgcgggca cggtccaggg gctggcgcct 4440
gtggcgtcct cggccgaccg cgacccgctg ggcccgcacc aggtacggat cgaggtgcgt 4500
gcggccgggc tgaacttccg cgatgtcctg gtggccctgg ggatggtccc tgggcagcgg 4560
gggctgggca gcgagggcgc cggggtggtg ctcgaagccg ggcctgaagt ggccgacctg 4620
gcgcccgggg accgggtgat gggcgtgttc gcggatgcgt tcggcccgtt cgcgatcgcc 4680
gaccgggcca ccgtgatccg cgtccccgac cactggacct tcggccaggc cgccgccgtc 4740
cccgtcgtgt tcgccaccgc ctattacggg ctggtggacc tggcaggact gcgcccgggt 4800
gagtcggtgc tggtgcacgc tgcggccggc ggagtgggac tggccgctgt ccaactggcc 4860
cgccacctgg gcgctgaggt ctacgccacg gcgagccccg gcaaatggga caccctacgc 4920
gcccacggca tccccccgga gcgcatcgcc tcgtcccgca ccctcgactt cgagagccgg 4980
ttcaccggcc ggaacatcga cgtcgtcctc aactccctgg cccatgagta cgtcgacgcc 5040
tcgctgcgcc tggtgtccgg cgacagcggc cggttcctgg agatgggcaa gaccgacctc 5100
cgcgacccgg aggaggtggc gcaggcgtac cccggtgtcg cctaccgggc gtacgacctg 5160
atggaggccg gacccgagcg catcggggag atcctgcgca ccgtgttgcg gctgttcgac 5220
gagggcgtgc tcaccccgct gccgctcacc tgctgggaca tccggcaggc cagggatgcc 5280
ttccgccaac tccagcaggg ccgcaccgtc ggaaagaatg tgctcacgct ggaccgcacc 5340
cccgaccccg acggcaccgt cctcatcacc ggtggcaccg gtaccctcgg cgccgcgctc 5400
gcccgccatc tcgccgccac cggccgagca cggcatctgc tactgatcag ccgccgtggc 5460
ctcgatgcgc caggcgctcc cgaactcatc gctgagattg acgagttggg cgccacggcg 5520
accgtcgcca cctgcgacgt cggcgaccgt gccgcgctcg ccgaactgct cgggcggatc 5580
cccgccgagc acccgctgac cgccgtcgtc cacgccgcgg gcaccctcga cgacgccacg 5640
ctcggctccc tcaccgcgcg ccacctcgac accgttctgc ccgcgaaggc cgatgccgcc 5700
tggcatctgc acgacctgac ctgccggctg gatctggccg cgttcgtgct gttctcgtcc 5760
gccgcgggtg tcctgggctc gccggggcag ggcaactacg ccgccgccaa cgcctttctc 5820
gacgcgctcg ccttccagcg acgggcgatg ggactccccg ccgtgtccct ggcatgggga 5880
ctgtgggagg aggccagcgg aatgaccggc cacctcgacc agaccgaccg cacccgcatg 5940
gcccgcgtcg gcctccggcc actggccacg gacgaggccc tggcgctgtt cgacaacgct 6000
ctcgtcgacg gcccaccgct gctgctcccg gcccgtatcg acaccaaggc gctacggggc 6060
accaccgcac cgcccctgtt ccagagcctc gtacgcccca ccaccggcca ccggccacgc 6120
cccgcgacac ccgacggccg ctcctccctc cgagcccggc tcgccgggct cgaccccgcc 6180
gcacagcacg aggtcctgct caccctcgtc cgcggccacg ccgccacggt cctcggccac 6240
ccgagccccg acgccatcgc ccgcgaggcg gccttccgtg acctcggctt cgactccctc 6300
accgccgtgg agctccgcaa ccgcctcaag gaggcaaccg gcctgcggct ccccgccacc 6360
atcgtcttcg accatcccac tcctgccgct ctcgcccagc acctgcggga cggcctcatc 6420
ggcggcgccg atacggtcac cctggctgcg gctcctgctc cgagcaaggt ggcgatggtg 6480
gcggatgagg ccatcgcgat catcggcatg gcctgccggt atccgggggg cgtgcggtcg 6540
gccgaggggc tgtgggatct ggtcgcctcc ggcaccgacg ccatgagcgg attccccagc 6600
gaccgcggct gggacctcga ccgcctctac gccccccagg accaggacgt gccgggcacc 6660
acatacaccc gccacggggg cttcctccac gacgcgggca agttcgacgc gggattcttc 6720
ggcatcggcc cacgtgaggc gctggcgatg gatccgcagc agcggctgct gctggagacc 6780
tcctgggagg ttttcgaaca cgcgggaatc gacccctcgt cggtacggcg gagccggacc 6840
ggagtcttcg ccggtgtgat gccgacggac tacggccccc ggctgcaaga caccgtggcc 6900
gaggtcgagg gctatgtcct caccggaaac tccggcagcg tcgcctcggg ccgtatcgcc 6960
tacaccttcg gcctggaagg ccccgcggtg tcggtggaca cggcgtgttc gtcgtctctg 7020
gtggcgttgc atctggcgtg tcaggcgctg cgtgcggggg agtgctccat ggcgctggcc 7080
ggcggggtga cggtgatggc gacgcctggt gccttcgtgg agtttgcgcg gcagcggggg 7140
ttgtcggtgg atgggcggtg caaggcgttt ggggtgggtg cggatggtac ggggtgggcg 7200
gagggggtgg ggatgctgtt ggtggagcgg ttgtctgatg cgcggcggtt ggggcatcgg 7260
gtgttggcgg tggtgcgggg ttctgcggtg aatcaggatg gtgcgtcgaa tggtttgacg 7320
gcgccgaatg gtccgtcgca gcagcgggtg atccggcagg cgttggccag tgcgcgggtt 7380
ggcggggcgg atgtggatgt ggtggagggg cacggtacgg ggacgcggct gggtgatccg 7440
atcgaggcgc aggcgttgct ggcgacctac ggtcaggagc gggtggggga cggctcgttg 7500
tggttggggt cggtgaagtc gaatatcggg catgcgcagg ccgcggcggg ggttgcgggt 7560
gtcatcaaga tggtgatggc gatgcggtat ggggtgttgc cgcggacgtt gcatgtgcag 7620
gagccgtcgc cgcatgtgga ctggtcctcg ggcggggtgc ggctgctgac ggaggcggtg 7680
ccgtggccgg agacggggcg tgcgcggcgt gcgggggtgt cgtcgttcgg ggtcagtggc 7740
accaacgcgc acatcatcct cgaacaggcg ccgcctgagg agcacgacga tccggcggac 7800
gtctcgtccg ggtcgtttcc gtggatggtg tcggccaagt ccgaacaggc actacaggcg 7860
caggcagcac agttgcgcgc gtatctggcg gcacatcctg agctggggct ggctgatgtc 7920
gggtatgcgc tggcctccgg ccgcacggcc ttcggccacc gtgccgtgct cctgggcccg 7980
gaccgcgaag ccttcgtcga agagctggga gctctggagg ccggtgagga acacgccggg 8040
ctggtacggg gcgtggcgac gggtgcgggg aagctggcgt ttgtgtgttc cgggcaggga 8100
acgcaacgtc cccgtatggg acacgggctg tactacgcct tcccgctgtt cgccgcagcc 8160
atggacgaag cctgcgcaca cctggaccca cacctcgacc atcccctgcg ggatgtcatg 8220
ttcgccgagc cgggcaccga caccgcccag ctgctccacc agacccgcta cgcccagccc 8280
gccctgttcg ccctccagat cgccctgcac cgcctggtca ccgaacacca cggccttacc 8340
ccccactact acgccggcca ttccctcgga gagatcaccg cggcccacct cgccgggatc 8400
ctcaccctcc ccgacgcggc ccgcctggtc accacccgcg cccgcctcat gcaatctctc 8460
cccgccaccg gcgccatgac caccctccaa gcagaccccg acgaactcca cgaacacctc 8520
acacgatgcg aaggacgggt ctcactcgcg gccgtgaacg cgcccgggtc cgtggtcatc 8580
agcggtgatc gccacgacgt agacgctacg gccgaaaacc tccgcgccat gggacgcaag 8640
accactgcgc tgaaggtcag cggcgctttc cactcacacc acatcgaccc actcctcaac 8700
gaactccgca acacggcaga aaccctcacc taccacccac cccacacccc cctcatcacc 8760
accaacccca ccgaccacga ccccaccaca ccccactact gggtccggca agcgcgcgag 8820
acggtccact acgcccacac cacccaacaa ctccacaccc acggcgtcac cgcctacctc 8880
gaactcggcc ccgaccacac cctcaccgcc ctcacccacc acaacctccc cgaccacacc 8940
ccgctagccg tcccgcttct ccaccccgac caatccgaga cccacaccac ccacaccgcc 9000
ctcgcccacc tccacaccca cggccacccc accacctggc accaccatca cacccccacc 9060
cactaccacc caaacctccc cacctacccc ttccaacacc accactactg gctcaacacc 9120
accactgcca ccggtgatat gtcggctgca ggccttgagc cggcgcggca tcccctgttg 9180
ggcgcggcgg tcgggttggc cgatggtgag gggttgctgt tcactgggcg gatttctctc 9240
cgtacgcatc cctggctggc cgaccacgcc gtcggcggcg ccgtgttgct ccccggtacg 9300
gcctttctcg aactcgccct ccaagccgcc gcccatgccg actgccgtcg ggtcgaggag 9360
cttacgctcc acaccccgct cgtcgtaccg gatagcgccg gcgtagtgct gcaggtcact 9420
gtggccgcgc cgaacgaagc aggaaaccgg gcggtggata tctactcgcg aatcgatgtc 9480
ggcggcctca ccgccgattc ggctggcgag ccgtggacgc gccatgccgc cgggtacctt 9540
gccgacaagc ctgacccaga ctgcggtgac tcggcggatg gtgtcatgcc cgcgggcgca 9600
tggccgccgc cgggtgcggt cgccgtggat ctggagggac tgtacgagca actggccgag 9660
gggggtttcc actacggtgc ggccttccgt tgcctggacg ccgcctggca acgcggggac 9720
gaggtcttcg cgaccgcgta tatgtcagag gatcagctgg gcgacacggc tgcggctcgg 9780
ttcgcgctgc accccgcgct gctggattcc gcactgcaca ccattccact tttgccctcc 9840
ctacggggac aacaggacag cgggctgccg ttcacgtgga caggagtcac cctgcgtgca 9900
tccggggcga cggctctgcg cgtccggctg aggccggacg gccatggccc gggggcggtg 9960
tcggtcgacg tgtccgacga ggcgggtgag cccgtagcat cggtccggtc gttggccctg 10020
cggccggtga ccagggccga gttgcatacg gccgagttgc gcacagccgc cccggttgcc 10080
ccccatggct cgctcttcga ggtgcgatgg gaacccgtcc cccagccttc agcggccgaa 10140
gaagccgccc catgggtgat gatcgggacc gggccgacgc tgcgcccggt cgaggacttc 10200
gtcactccgc cggagcggac gtacgccgac ctggccgcgc tgtgcgtggc aatcgccgat 10260
gacgcgcccg ttccccggac ggtcgtggcc tggtccccag ccgggagcga agacgagtcg 10320
agtgaggcgc tgcgccaggc cacacaccac atgctgggcc tactgcagca gtggttggcg 10380
gacagccggt tcgccgacag tcgcctggtg atcctcaccc gagccgcggt ggccactgcg 10440
ccggacgagg aggtagaaga cctggcggga gcggcggcgc ggggtctgat ccgctccgcc 10500
cagtcggagc accctgaccg attcgtcctg ctcgacctgg acgaccgtcc cgctgacgcg 10560
aaagaccacg accgaatgct gtcgatggcc ctggcctgcg gggaaccgga agtggccgta 10620
cgcgatggag ccctgcgcac accccggctg agcccgctgg ccggcaccgc caccgaggcc 10680
atggacgagc atccctggga tcaggacggc accgtactca tcaccggcgg caccggcagc 10740
ctcggcgcca tgcttgcccg ccacttggtg gccacccatg gcgtacggca tctgatgctg 10800
atcagccgac gtggcctcga cgccccgggg gccaggcgac tgggggtcga acttgcggag 10860
ctcggggcgc aggtgacgat caccgcgtgc gatgccgcag accaaaggca acttgcgaac 10920
gtattgtcgg agatctccgt cgaccatccg ctgaccgctg tggtgcatgc ggcaggcgta 10980
ctggacgacg gggtgatcac atccctcaca ccggagggcc tgacccatgt cctgcgggcc 11040
aaggtcgatt cggcgctcaa tctccaccag ctcacacgcg acctgccgct gtccgcgttt 11100
gtgctcttct cctcgctggc cggggtgatg ggttcggcag ggcagggcaa ctacgccgcc 11160
gccaacgcag ccctggacgc gctggcgagt caccggaggg ccgctcggct gccggcggtg 11220
tccttggcct ggggagtttg ggagcagacc gagggcatga ccgggcagtt ggaggccacg 11280
gaccacgcgc ggctccgccg ctcgggcctg aggccgctgg ccatcagcga gggcctggag 11340
ctcttcgaca aggccctgag ctgtggacac gccctggtgg tgcccgccgc actcagcacg 11400
agggagcttc agacatccgg atccgtcccg ccattcctgc gccacctgac gggtgtcgct 11460
ccggcccggc cgtcccggac ccgcgacgcc tcggccggtg agccgacctc cctgcggcgg 11520
cggttgaccg gcctcgggcc ggaagaacgg ctacgcgagg tgctgcggct ggtgcgctcc 11580
cgggcggctg cggtgctggg gcacggcacg gccgaatcgg tcccggcgga ctcggcgttc 11640
cgcgacctgg ggttcgactc cctcgccgcg gtggacctgc ggaaccggtt gcagcaggcc 11700
accgggctgc gcctgccggc cggcttgatc ttcgaccggc cgcgtccgga cgtgctcgcc 11760
cgtttcctgt gtgacgagtt ggccggcgcc ggcggtacgt cggcggccac ggccgcccca 11820
cccgttgcgg ccgtcggcgg ggcagccggc gagccggtgg ccatcgtcgg catggcatgc 11880
cggtttccgg gaggtgtgcg gtcggccgag ggcctgtggg atctggtcgc ctccggtatg 11940
gacgcggtgg gtgacttccc cgcagaccga ggctgggagg tggaacggct ctacgacccc 12000
gacccggacc gaaccggcac ctcctacacc cggcaaggcg ggttccttta cgacgcgggt 12060
gagttcgacg cggcattctt cgggatcggc ccgcgtgagg cggtagccat ggatccacag 12120
cagcggctgc tgctggagat ctcctgggag gcgctggaac gtgcggggat cgacccggcg 12180
tcgctgcggg ggagttcgac cggggtgttc gctggggtga tgtaccacga ctacggcacc 12240
cgcctgcgcg agatcccaga gggctacgag ggctatatcg gcaatggaaa cgcgggcagc 12300
gtcgcgtcgg gacgtgtcgc ctacaccttc ggcctggagg ggccggcggt caccgtggac 12360
acggcgtgtt cgtcgtccct ggtcgccctg catctggcct gccaggcgct gcggtcaggg 12420
gagtgctcca tggcgctggc cggcggggtc accgtcatgt ccacccccac cacttttgtc 12480
gagttctcgc gccagcgggg actggccccg gacgggcggt gcaagtcctt cggggccggc 12540
gcggacggaa caggctgggc ggagggggcg gggatgctcc tggtggaacg gctttcggac 12600
gcccggcgca acggccaccg ggtcctggcg gtggtacggg ggagtgcggt caaccaggac 12660
ggggcgagca atgggctgac ggcgccgaac ggcccgtcgc aagagcgggt gatccgccag 12720
gcgtgggcaa acgcgggtgt ggccgcgatg gacatcgacg cggtggaggg acacggcacg 12780
gggacgacgc tcggtgaccc catcgaggcc caggcgctgc tggggacgta cggacaggga 12840
cggtcggccg atcggccgtt gtggttggga tcgatcaagt ccaacgtcgg acacacccag 12900
gccgccgcgg gggtgggcgg cgtcatcaag atggtgatgg ccatgcgcca cgggctgctc 12960
ccgcagaccc tgcacgccga ggagccctca cctcatgtgg actggtcggg cgggacggtg 13020
cggttgctga ccgagtcggt ggcctggccc gagcaggggc ggatgcgccg tgcgggcgtc 13080
tcctctttcg gtgtcagcgg taccaacgcc cacgtcatcc tggaacaagc accgcctgcc 13140
gcggagaccc acgaaccggc agagcccaac accgcgccag gcccactgcc ctgggcgatc 13200
tccgcgaaga gcccgcaagc gctacgtgcc caggcgcgcc aactgcacac gtacctgacc 13260
aacgcccccg aggcgaaccc cgccgacgtc ggccacaccc tcgcgacggg ccgcgcctct 13320
ttcgagcatc gtgctgtggt catcggctcc gaccgagcgg agttcctggg tggcctggat 13380
gctctggcgg ccgacgaggc ccacaccgcc gtcgtcacgg ggatcgcgag gaaggccggt 13440
gaccagggca aggtggtgtt cgtgttcccc gggcagggcg gtcagtgggc cgggatggga 13500
ctgcggctgc ttaagacctc acccgtcttc gcccaatcga tccaggcctg cgaacaagcc 13560
ctcgcccccc acaccgactg gaccctgacc gacatcctgc accggcccca caccgacccc 13620
ctgtggcagc gcgccgacgt catccagccc gtcctcttcg ccctcatgac ctccctcgcc 13680
gccctctggc aatcccacgg ccttaacccc gacgccgtca tcggccactc ccaaggcgaa 13740
atcaccgccg cccacatcag cggagcgctg agcctggagg acgccgcgaa aaccgtcgcg 13800
ctgcgcagcc gggccctgca gaccctgcgc ggttcgggcg gcatggcctc cgtaccactg 13860
ccggcggacc aggtcaccgg gctgctgcag accatgtggc cggaccggct gtgggtggcc 13920
gccgtcaacg cccctaccgc cacggtgatc tccggcaacg cggaagctct cacacaggcg 13980
ctggaacact accgggacca aggcgtcgac gcgaaacgga tcccggtcga ctacgcctcc 14040
cactgccccc acatccaggc cgtggaacag gaactgtcac ggctgttgcg gggcatcacc 14100
ccacgggccg ccaccacccc cttctactcc accaccgaca accaatggac cgacaccacc 14160
accctcaacg cccactactg gtaccgaaac ctccgccaac ccgtccacct cgccgacgcc 14220
atcaccaacc tcacccacca aggccaccac accttcatcg aaatcagccc ccaccccacc 14280
ctcacccccg ccatccaaga aaccaccgac accacccaca cccccaccac cgtcatcagc 14340
acactccgcc gcaaccacaa cgacacccac caaatcctcc acgccctcgc ccacgcccac 14400
accaccggcc accccatcaa ctggcacacc acccaccaac accacacccc aaccccccaa 14460
cacatcgacc tacccaccta ccccttccaa caccaccact actggctcaa cacccccacc 14520
cagacagggg atgcggcggc cgtcggcctg gacccggcac atcacccgtt gctgggcgcg 14580
gcggtcgcgg tggccgaggg ggagggctat ctgctcaccg gtcggctcgc cctgtccacc 14640
cacccctggc tcgccgatca caccatcgcc ggcgcggttg tcctccctgg aactgccctt 14700
ctcgagatcg cccttcaggc gggccatcgt gtggactgct ggcgcatcga agaactcacc 14760
ctccaatcac cgctgttcat cccggaagag ggagcagtac aggtgcaggc atgggtggcg 14820
gcaccggatg agaacgggtg ccgaagcctg acggtgtcct cccgacgcga gggtacgtac 14880
gaggacgcca cgtgggtgcg ccatgccacg ggccgggtcg gccccgcacc ggccgaccag 14940
gatgaagcca tcgcacggct caccgaccca caaggcgacg gagcggcggc ggcggtctgg 15000
ccaccgcagg gcgctgtcgc gttcaccgca gacgatctgg agggcctgta cgacgggtac 15060
gcggcgcggg gattcgagta cggcccggtg ttccgaggcc tgcgggcggc ctggcgacgt 15120
ggcgaggaca tcttcgccga ggtgcgcctt cccgacacgg cggacggcga cgcctcccag 15180
ttctccgtac accccgccct gctggacgcc gcactgcacg ccgcggcctt ccgcccggcc 15240
gacaaactcc cgcacggcgc cctgccgttc tccttcagcg gggtgaggct gcacgggccc 15300
ggagcgtcga ccctgcgggt gcgcctcacc ccggacggcc aggcgcggga cacgcacgca 15360
tggtcggtcg cggtggtcga cggcgagggg cggccggtgg cctcgatcgc atcgctcgcg 15420
gtccgcccgg tgtcgacgca ggagttgctg gcggcctccg gtacggcgcg gcgggactcg 15480
ctcttcgcgg tcgagtgggt gaccgccctg gcgccgacct cgtcgtccgt tccgcaacgc 15540
ctggccacgg tggggcccag cgaccgcctc ccctcggcag acgcgtacgc gaacctcgcc 15600
gacctggccg ccgcagtgct ggaggcgggg gccccggcgc ccgatgcggt cgtggtcgac 15660
tgcggccgcc gcgatgcgcg cgccaccgcc gtgccggagg acgtaaggac cctcacccgg 15720
cgcatcctgg gtctgctgca ggagtggctg gcggacgaga ggccggcctc gagccggatg 15780
gtcgtactga cccgtggtgc ggtggccacc actccggggg aggacgtggc ggacctggcg 15840
ggcgcggcgg tgtgcggcat ggtgcgctcc gcgcagtcgg aacatcccgg ccggttcgtc 15900
ctgctggacc tcgaccccga cccggacctc gacggcgggg aagtgccacc gaccgtcgtt 15960
ccggcggctc tcgcctgtgg tgagccgcag atcgcggtgc gtgcgaaccg gcacctggtg 16020
ccccggctga cccgcgttcc ggcgtccgtc cccgtccccg ggcgtgttcc cgttcccgcc 16080
gccgaggcag ccgacccgga caccacgccc acggcgttcg accccgacgg caccgtagtg 16140
atcaccggcg gcaccggcac ccttggcgcg atgctcgcgc gccatctggt cagccgtcac 16200
ggtgtacgac acctcctgct ggcatcgcga cgcggacccg acgcacccgg cgccaccgag 16260
ctgcgggcgg aactggccga gctcggcgcc gaggtgacgg tgcgcgcttg tgacaccggt 16320
gaccgaggcg cgctggcgga tctcatcgcg gggattccca ccggccaccc tttgaccggt 16380
gtggtccacg ctgcgggcgt cctggacgac gccaccgtcg cctcgctcac cccccgacac 16440
ctggacaccg cgctgacacc caaggccgac gccgccttcc atctgcacga gctcacccgc 16500
cacgcccggc cgcgcgcctt cgtcctgttc tcctcggccg ccggtgtcct cggcgcagcc 16560
gggcagggca actatgcggc cgccaacgct ttcctcgacg ccctcgccga acaccgcagg 16620
gcgcagggcc tgccggcctt gtcgctcgcg tggggcctgt gggagcaggg cagcggcatg 16680
accgggcatc tcgaccgcac cgaccgggcc cgcatcaacc gctccggact cgcccccctc 16740
gccacggagg acgctctcgc gctcttcgac gccgccctcg ccggcgatcg gccgttcctg 16800
gtgcccgccc ggctggacct gcggggttca agcgccgccg agaccccggc gccgctgttc 16860
tccaggatcg ccccggctcg tacgacccgg ggccggtccc ccggcgccga gggcgccgct 16920
gaccttcgta cccgtctcgc ggcccaggac gccgccgagc agcgcgacac gcttctcacg 16980
atcgtccgca cccacaccgc cgccgtcctg gggcatgaca cggctgccgc cgtgcggccg 17040
gacggggcct tccgtgaact gggtttcgac tccctcgccg ccgtggaact ccgtaaccgc 17100
cttcaaacga ccaccgccct caccctgccc gcgaccaccg tcttcgacca ccccaccccc 17160
gctgccctcg ccgatcatct gcgtactcag ctctgccagg acgctcagtc ctcggcggcg 17220
gccacggcca tggcggcgat ggcggagctg gccaggctgg agtccgccgt ctccgattcg 17280
gtggcgctcg acgacgacac gcgcagcggc ctcgcggagc gcctgcggtc cctcgcccgc 17340
aagatgagca gtggccgtgt cgtcgaccac gacggcggcg gcgctgcgga cctggatctt 17400
cagtcggtca cggacgatga gatgttcgag ctgatcgaca aggaggtcag ccgagactga 17460
17460
<210> 74
<211> 5819
<212> PRT
<213> Artificial Sequence
<220>
<223> milA3 protein of Streptomyces milbemycinicus
<400> 74
Met Ala Ala Gly His Asp Lys Val Ile Glu Ala Leu Arg Ala Ser Leu
1 5 10 15
Lys Thr Asn Glu Arg Gln Arg Glu Gln Ile His Arg Leu Thr Thr Ala
20 25 30
Ala Arg Glu Pro Ile Ala Ile Ile Gly Met Ala Cys Arg Tyr Pro Gly
35 40 45
Gly Val Gly Ser Pro Glu Asp Leu Trp Glu Leu Val Ala Ala Gly Arg
50 55 60
Asp Ala Ile Gly Thr Phe Pro Glu Asp Arg Gly Trp Asp Val Glu Arg
65 70 75 80
Leu Tyr Asp Pro Asp Pro Glu Arg Ala Gly Thr Ser Cys Thr Gln His
85 90 95
Gly Gly Phe Leu Tyr Gln Ala Gly Glu Phe Asp Pro Gly Phe Phe Gly
100 105 110
Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu
115 120 125
Leu Glu Ile Ser Trp Glu Val Phe Glu Arg Ala Gly Ile Asp Pro Ala
130 135 140
Ser Val Arg Gly Ser Arg Thr Gly Val Phe Ala Gly Val Met Tyr His
145 150 155 160
Asp Tyr Gly Ser Arg Leu His Thr Val Pro Glu Gly Phe Glu Gly Tyr
165 170 175
Val Gly Asn Gly Ser Gly Gly Gly Val Ala Ser Gly Arg Val Ala Tyr
180 185 190
Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser
195 200 205
Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ala Gly
210 215 220
Glu Cys Ser Leu Ala Leu Ala Gly Gly Val Thr Val Met Ser Thr Pro
225 230 235 240
Ser Leu Phe Val Glu Tyr Ser Arg Gln Arg Ala Leu Ala Ala Asp Gly
245 250 255
Arg Cys Lys Ala Tyr Gly Ala Gly Ala Asp Gly Thr Gly Trp Ala Glu
260 265 270
Gly Ala Gly Met Leu Leu Val Glu Arg Leu Thr Asp Ala Gln Arg Leu
275 280 285
Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp
290 295 300
Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala Gln Gln Arg
305 310 315 320
Ala Ile Arg Gln Ala Leu Ala Ser Ala Gly Val Ser Ala Ser Glu Val
325 330 335
Asp Ala Val Glu Gly His Gly Thr Gly Thr Arg Leu Gly Asp Pro Ile
340 345 350
Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Gln Arg Pro Ala Asp
355 360 365
Arg Pro Leu Trp Leu Gly Ser Met Lys Ser Asn Val Gly His Ala Gln
370 375 380
Ala Ala Ala Gly Val Gly Gly Ile Ile Lys Met Val Met Ala Met Arg
385 390 395 400
Ser Gly Thr Leu Pro Arg Thr Leu His Ala Asp Glu Pro Ser Pro His
405 410 415
Ile Asp Trp Asp Ser Gly Ala Val Arg Leu Leu Thr Glu Pro Val Ala
420 425 430
Trp Pro Glu Arg Asp Arg Pro Arg Arg Ala Ala Val Ser Ser Phe Gly
435 440 445
Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Ala Ala Ser Gln Thr
450 455 460
Ala Pro Gln Thr Asp Ser Ala Ser Gln Ala Glu Thr Asp Asp Ala Pro
465 470 475 480
Ala Pro His Gly Ala Pro Gly His Ala Val Ala Gly Pro Leu Leu Trp
485 490 495
Pro Leu Ser Gly Ala Thr Ala Glu Ala Leu Arg Ala Gln Ala Gly Glu
500 505 510
Leu Arg Arg Phe Val Ala Ala Asp Glu Leu Leu Arg Pro Ala Asp Val
515 520 525
Gly His Thr Leu Val Phe Gly Arg Ser Asp Leu Ala His Arg Ala Val
530 535 540
Val Leu Gly Ser Asp Arg Glu Thr Leu Leu Arg Ala Leu Asp Thr Leu
545 550 555 560
Ala Gly Glu Gly Pro Asp Asp Gly Ser Val Val Arg Gly Met Ala Ala
565 570 575
Ala Gly Ala Gly Ala Gly Val Val Phe Val Phe Pro Gly Gln Gly Gly
580 585 590
Gln Trp Ala Gly Met Gly Leu Arg Leu Leu Glu Thr Ser Ser Phe Phe
595 600 605
Ala Glu Arg Met Ala Glu Cys Glu Ala Ala Leu Ala Pro Tyr Ala Asp
610 615 620
Trp Ser Leu Leu Asp Val Leu Arg Arg Asp Pro Gly Asp Pro Val Trp
625 630 635 640
Glu Arg Ala Asp Val Val Gln Pro Met Leu Phe Ser Val Met Val Ser
645 650 655
Leu Ala Gln Leu Trp Arg Ser Tyr Gly Val Glu Pro Asp Ala Val Leu
660 665 670
Gly His Ser Gln Gly Glu Ile Ala Ala Ala His Ile Cys Gly Ala Leu
675 680 685
Thr Leu Asp Asp Ala Ala Lys Val Val Ala Leu Arg Ser Arg Ala Leu
690 695 700
Gln Thr Leu Arg Gly Ser Gly Gly Met Ala Ser Val Pro Leu Pro Ala
705 710 715 720
Asp Glu Val Thr Gly Leu Leu Arg Thr Ala Trp Pro Asp Arg Leu Trp
725 730 735
Val Ala Ala Val Asn Ala Pro Thr Ala Thr Val Ile Ser Gly Asp Ala
740 745 750
Asp Ser Leu Ala Glu Ala Leu Glu His Tyr Arg Asp Gln Gly Val Glu
755 760 765
Ala Lys Arg Val Pro Val Asp Tyr Ala Ser His Cys Pro His Ile Glu
770 775 780
Ala Val Glu Gln Glu Leu Leu Gly Leu Leu Arg Gly Ile Ala Pro Arg
785 790 795 800
Ala Ala Asp Ile Pro Phe Tyr Ser Thr Val Asp Asn Gln Trp Ala Asp
805 810 815
Thr Met Gly Leu Asp Ala Arg Tyr Trp Tyr Arg Asn Leu Arg Arg Pro
820 825 830
Val Arg Phe Ala Glu Ala Leu Arg Ala Leu Gly Ala Ala Glu Tyr Arg
835 840 845
Thr Tyr Val Glu Val Gly Pro His Pro Thr Leu Thr Pro Ala Ile Glu
850 855 860
Asp Thr Thr Glu Ala Ala Gly Val Ala Ala Thr Val Val Gly Ser Leu
865 870 875 880
Arg Arg Gly Glu Asp Asp Ala His Arg Ile Leu Thr Ser Leu Ala Arg
885 890 895
Ala His Ile His Gly Leu Pro Val Ala Trp Asp Arg His Tyr Arg Ala
900 905 910
Leu Ala Pro Glu Ala Asn His Val Asp Leu Pro Thr Tyr Ala Phe Gln
915 920 925
Arg Arg Arg Tyr Trp Leu Asp Ala Pro Ala Thr Thr Gly Asp Val Thr
930 935 940
Ala Ala Gly Leu Ala Pro Val Gly His Pro Leu Leu Gly Ala Ala Val
945 950 955 960
Gly Leu Ala Glu Gly Asp Gly Tyr Leu Leu Thr Gly Arg Leu Ala Pro
965 970 975
His Thr His Pro Trp Leu Thr Asp His Ala Val Ala Gly Thr Val Leu
980 985 990
Leu Pro Gly Thr Ala Tyr Val Glu Leu Ala Val His Val Gly Gly His
995 1000 1005
Leu Gly Cys Pro Arg Leu Glu Glu Leu Thr Leu His Ala Pro Leu Val
1010 1015 1020
Leu Pro Asp Thr Gly Gly Val Ala Leu Gln Val Ala Val Gly Ala Pro
1025 1030 1035 1040
Asp Glu Thr Gly Arg Arg Ala Leu Ser Val Tyr Ala Gln Arg Asp Asp
1045 1050 1055
Asp Pro Ala Trp Glu Gly Ala Ala Arg Gly Ala Trp Thr Arg His Ala
1060 1065 1070
Thr Gly Thr Leu Ala Ala Glu Ala Pro Thr Asp Gly Ile Ser Gly Ala
1075 1080 1085
Asp Gly Ala Gly Thr Leu Ala Gly Ala Trp Pro Pro Pro Gly Ala Glu
1090 1095 1100
Pro Leu Asp Ile Ser Gly Leu Tyr Asp Thr Leu Ala Ala Ala Asp Phe
1105 1110 1115 1120
Gly Tyr Gly Pro Ala Phe Gln Gly Leu Arg Ala Val Trp Arg Gln Gly
1125 1130 1135
Glu Glu Thr Tyr Ala Glu Val Arg Leu Pro Asp Gln Val Ala Ala Asp
1140 1145 1150
Ala Pro Arg Phe Cys Leu His Pro Ala Leu Leu Asp Ala Ala Leu His
1155 1160 1165
Pro Leu Ala Leu Asp Ser Gly Arg Ser Glu Glu Asn Pro Ala Gly His
1170 1175 1180
Gly Leu Leu Pro Phe Ala Trp Arg Gly Val Ser Leu Arg Ser Pro Gly
1185 1190 1195 1200
Thr Pro Thr Leu Arg Val Arg Leu Arg Pro Gln Gly Pro Asp Ser Ile
1205 1210 1215
Ala Val Asp Val Ala Asp Glu Thr Gly Ala Pro Val Ala Ser Ala Glu
1220 1225 1230
Ser Leu Thr Leu Arg Pro Val Ala Leu Glu Asp Leu Arg Ala Leu Gly
1235 1240 1245
Gly Gln Ala Gly Asp Thr Leu Tyr Ala Leu Glu Trp Thr Ala Ala Pro
1250 1255 1260
Glu Pro Pro Ala Thr Ala Leu Gly Arg Cys Ala Val Ile Gly Gln Ala
1265 1270 1275 1280
Ile Pro Gly Trp Ala Ala Ala Leu Glu Thr Ala Ala Ala Gly Pro Val
1285 1290 1295
Arg Arg Tyr Pro Asp Leu Ala Gly Leu Val Thr Ala Leu Asp Ala Gly
1300 1305 1310
Asp Pro Pro Pro Asp Leu Val Phe Val Gly Cys Pro Pro Ala Ala Ala
1315 1320 1325
Gly Pro Asp Asp Thr Thr Val Ala Asp Val His Thr Ala Arg Thr Arg
1330 1335 1340
Val Arg Thr Arg Gln Ala Leu Asp Leu Leu Gln Gly Trp Leu Gly Glu
1345 1350 1355 1360
Ala Arg Leu Ala Gly Ala Arg Leu Val Leu Val Thr Cys Gly Ala Val
1365 1370 1375
Ala Thr Gly Pro Ala Glu Gly Val Met Asp Leu Ala Gly Ala Ala Ile
1380 1385 1390
Cys Gly Leu Val Arg Ser Ala Gln Ala Glu Glu Pro Asp Arg Ile Leu
1395 1400 1405
Leu Val Asp Leu Asp Ala Ala Glu Glu Ser Trp Ala Ala Leu Pro Arg
1410 1415 1420
Ala Val Ala Leu Gly Glu Pro Gln Met Ala Ile Arg Ala Gly Gln Pro
1425 1430 1435 1440
His Met Ala Arg Leu Val Arg Ala Asp Thr Glu Gly Gly Ala Leu Leu
1445 1450 1455
Thr Pro Pro Gln Gly Ser Gly Gly Trp Arg Leu Asp Cys Ala Asp Ala
1460 1465 1470
Gly Thr Val Gln Gly Leu Ala Pro Val Ala Ser Ser Ala Asp Arg Asp
1475 1480 1485
Pro Leu Gly Pro His Gln Val Arg Ile Glu Val Arg Ala Ala Gly Leu
1490 1495 1500
Asn Phe Arg Asp Val Leu Val Ala Leu Gly Met Val Pro Gly Gln Arg
1505 1510 1515 1520
Gly Leu Gly Ser Glu Gly Ala Gly Val Val Leu Glu Ala Gly Pro Glu
1525 1530 1535
Val Ala Asp Leu Ala Pro Gly Asp Arg Val Met Gly Val Phe Ala Asp
1540 1545 1550
Ala Phe Gly Pro Phe Ala Ile Ala Asp Arg Ala Thr Val Ile Arg Val
1555 1560 1565
Pro Asp His Trp Thr Phe Gly Gln Ala Ala Ala Val Pro Val Val Phe
1570 1575 1580
Ala Thr Ala Tyr Tyr Gly Leu Val Asp Leu Ala Gly Leu Arg Pro Gly
1585 1590 1595 1600
Glu Ser Val Leu Val His Ala Ala Ala Gly Gly Val Gly Leu Ala Ala
1605 1610 1615
Val Gln Leu Ala Arg His Leu Gly Ala Glu Val Tyr Ala Thr Ala Ser
1620 1625 1630
Pro Gly Lys Trp Asp Thr Leu Arg Ala His Gly Ile Pro Pro Glu Arg
1635 1640 1645
Ile Ala Ser Ser Arg Thr Leu Asp Phe Glu Ser Arg Phe Thr Gly Arg
1650 1655 1660
Asn Ile Asp Val Val Leu Asn Ser Leu Ala His Glu Tyr Val Asp Ala
1665 1670 1675 1680
Ser Leu Arg Leu Val Ser Gly Asp Ser Gly Arg Phe Leu Glu Met Gly
1685 1690 1695
Lys Thr Asp Leu Arg Asp Pro Glu Glu Val Ala Gln Ala Tyr Pro Gly
1700 1705 1710
Val Ala Tyr Arg Ala Tyr Asp Leu Met Glu Ala Gly Pro Glu Arg Ile
1715 1720 1725
Gly Glu Ile Leu Arg Thr Val Leu Arg Leu Phe Asp Glu Gly Val Leu
1730 1735 1740
Thr Pro Leu Pro Leu Thr Cys Trp Asp Ile Arg Gln Ala Arg Asp Ala
1745 1750 1755 1760
Phe Arg Gln Leu Gln Gln Gly Arg Thr Val Gly Lys Asn Val Leu Thr
1765 1770 1775
Leu Asp Arg Thr Pro Asp Pro Asp Gly Thr Val Leu Ile Thr Gly Gly
1780 1785 1790
Thr Gly Thr Leu Gly Ala Ala Leu Ala Arg His Leu Ala Ala Thr Gly
1795 1800 1805
Arg Ala Arg His Leu Leu Leu Ile Ser Arg Arg Gly Leu Asp Ala Pro
1810 1815 1820
Gly Ala Pro Glu Leu Ile Ala Glu Ile Asp Glu Leu Gly Ala Thr Ala
1825 1830 1835 1840
Thr Val Ala Thr Cys Asp Val Gly Asp Arg Ala Ala Leu Ala Glu Leu
1845 1850 1855
Leu Gly Arg Ile Pro Ala Glu His Pro Leu Thr Ala Val Val His Ala
1860 1865 1870
Ala Gly Thr Leu Asp Asp Ala Thr Leu Gly Ser Leu Thr Ala Arg His
1875 1880 1885
Leu Asp Thr Val Leu Pro Ala Lys Ala Asp Ala Ala Trp His Leu His
1890 1895 1900
Asp Leu Thr Cys Arg Leu Asp Leu Ala Ala Phe Val Leu Phe Ser Ser
1905 1910 1915 1920
Ala Ala Gly Val Leu Gly Ser Pro Gly Gln Gly Asn Tyr Ala Ala Ala
1925 1930 1935
Asn Ala Phe Leu Asp Ala Leu Ala Phe Gln Arg Arg Ala Met Gly Leu
1940 1945 1950
Pro Ala Val Ser Leu Ala Trp Gly Leu Trp Glu Glu Ala Ser Gly Met
1955 1960 1965
Thr Gly His Leu Asp Gln Thr Asp Arg Thr Arg Met Ala Arg Val Gly
1970 1975 1980
Leu Arg Pro Leu Ala Thr Asp Glu Ala Leu Ala Leu Phe Asp Asn Ala
1985 1990 1995 2000
Leu Val Asp Gly Pro Pro Leu Leu Leu Pro Ala Arg Ile Asp Thr Lys
2005 2010 2015
Ala Leu Arg Gly Thr Thr Ala Pro Pro Leu Phe Gln Ser Leu Val Arg
2020 2025 2030
Pro Thr Thr Gly His Arg Pro Arg Pro Ala Thr Pro Asp Gly Arg Ser
2035 2040 2045
Ser Leu Arg Ala Arg Leu Ala Gly Leu Asp Pro Ala Ala Gln His Glu
2050 2055 2060
Val Leu Leu Thr Leu Val Arg Gly His Ala Ala Thr Val Leu Gly His
2065 2070 2075 2080
Pro Ser Pro Asp Ala Ile Ala Arg Glu Ala Ala Phe Arg Asp Leu Gly
2085 2090 2095
Phe Asp Ser Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Lys Glu Ala
2100 2105 2110
Thr Gly Leu Arg Leu Pro Ala Thr Ile Val Phe Asp His Pro Thr Pro
2115 2120 2125
Ala Ala Leu Ala Gln His Leu Arg Asp Gly Leu Ile Gly Gly Ala Asp
2130 2135 2140
Thr Val Thr Leu Ala Ala Ala Pro Ala Pro Ser Lys Val Ala Met Val
2145 2150 2155 2160
Ala Asp Glu Ala Ile Ala Ile Ile Gly Met Ala Cys Arg Tyr Pro Gly
2165 2170 2175
Gly Val Arg Ser Ala Glu Gly Leu Trp Asp Leu Val Ala Ser Gly Thr
2180 2185 2190
Asp Ala Met Ser Gly Phe Pro Ser Asp Arg Gly Trp Asp Leu Asp Arg
2195 2200 2205
Leu Tyr Ala Pro Gln Asp Gln Asp Val Pro Gly Thr Thr Tyr Thr Arg
2210 2215 2220
His Gly Gly Phe Leu His Asp Ala Gly Lys Phe Asp Ala Gly Phe Phe
2225 2230 2235 2240
Gly Ile Gly Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu
2245 2250 2255
Leu Leu Glu Thr Ser Trp Glu Val Phe Glu His Ala Gly Ile Asp Pro
2260 2265 2270
Ser Ser Val Arg Arg Ser Arg Thr Gly Val Phe Ala Gly Val Met Pro
2275 2280 2285
Thr Asp Tyr Gly Pro Arg Leu Gln Asp Thr Val Ala Glu Val Glu Gly
2290 2295 2300
Tyr Val Leu Thr Gly Asn Ser Gly Ser Val Ala Ser Gly Arg Ile Ala
2305 2310 2315 2320
Tyr Thr Phe Gly Leu Glu Gly Pro Ala Val Ser Val Asp Thr Ala Cys
2325 2330 2335
Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ala
2340 2345 2350
Gly Glu Cys Ser Met Ala Leu Ala Gly Gly Val Thr Val Met Ala Thr
2355 2360 2365
Pro Gly Ala Phe Val Glu Phe Ala Arg Gln Arg Gly Leu Ser Val Asp
2370 2375 2380
Gly Arg Cys Lys Ala Phe Gly Val Gly Ala Asp Gly Thr Gly Trp Ala
2385 2390 2395 2400
Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg
2405 2410 2415
Leu Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln
2420 2425 2430
Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln
2435 2440 2445
Arg Val Ile Arg Gln Ala Leu Ala Ser Ala Arg Val Gly Gly Ala Asp
2450 2455 2460
Val Asp Val Val Glu Gly His Gly Thr Gly Thr Arg Leu Gly Asp Pro
2465 2470 2475 2480
Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Glu Arg Val Gly
2485 2490 2495
Asp Gly Ser Leu Trp Leu Gly Ser Val Lys Ser Asn Ile Gly His Ala
2500 2505 2510
Gln Ala Ala Ala Gly Val Ala Gly Val Ile Lys Met Val Met Ala Met
2515 2520 2525
Arg Tyr Gly Val Leu Pro Arg Thr Leu His Val Gln Glu Pro Ser Pro
2530 2535 2540
His Val Asp Trp Ser Ser Gly Gly Val Arg Leu Leu Thr Glu Ala Val
2545 2550 2555 2560
Pro Trp Pro Glu Thr Gly Arg Ala Arg Arg Ala Gly Val Ser Ser Phe
2565 2570 2575
Gly Val Ser Gly Thr Asn Ala His Ile Ile Leu Glu Gln Ala Pro Pro
2580 2585 2590
Glu Glu His Asp Asp Pro Ala Asp Val Ser Ser Gly Ser Phe Pro Trp
2595 2600 2605
Met Val Ser Ala Lys Ser Glu Gln Ala Leu Gln Ala Gln Ala Ala Gln
2610 2615 2620
Leu Arg Ala Tyr Leu Ala Ala His Pro Glu Leu Gly Leu Ala Asp Val
2625 2630 2635 2640
Gly Tyr Ala Leu Ala Ser Gly Arg Thr Ala Phe Gly His Arg Ala Val
2645 2650 2655
Leu Leu Gly Pro Asp Arg Glu Ala Phe Val Glu Glu Leu Gly Ala Leu
2660 2665 2670
Glu Ala Gly Glu Glu His Ala Gly Leu Val Arg Gly Val Ala Thr Gly
2675 2680 2685
Ala Gly Lys Leu Ala Phe Val Cys Ser Gly Gln Gly Thr Gln Arg Pro
2690 2695 2700
Arg Met Gly His Gly Leu Tyr Tyr Ala Phe Pro Leu Phe Ala Ala Ala
2705 2710 2715 2720
Met Asp Glu Ala Cys Ala His Leu Asp Pro His Leu Asp His Pro Leu
2725 2730 2735
Arg Asp Val Met Phe Ala Glu Pro Gly Thr Asp Thr Ala Gln Leu Leu
2740 2745 2750
His Gln Thr Arg Tyr Ala Gln Pro Ala Leu Phe Ala Leu Gln Ile Ala
2755 2760 2765
Leu His Arg Leu Val Thr Glu His His Gly Leu Thr Pro His Tyr Tyr
2770 2775 2780
Ala Gly His Ser Leu Gly Glu Ile Thr Ala Ala His Leu Ala Gly Ile
2785 2790 2795 2800
Leu Thr Leu Pro Asp Ala Ala Arg Leu Val Thr Thr Arg Ala Arg Leu
2805 2810 2815
Met Gln Ser Leu Pro Ala Thr Gly Ala Met Thr Thr Leu Gln Ala Asp
2820 2825 2830
Pro Asp Glu Leu His Glu His Leu Thr Arg Cys Glu Gly Arg Val Ser
2835 2840 2845
Leu Ala Ala Val Asn Ala Pro Gly Ser Val Val Ile Ser Gly Asp Arg
2850 2855 2860
His Asp Val Asp Ala Thr Ala Glu Asn Leu Arg Ala Met Gly Arg Lys
2865 2870 2875 2880
Thr Thr Ala Leu Lys Val Ser Gly Ala Phe His Ser His His Ile Asp
2885 2890 2895
Pro Leu Leu Asn Glu Leu Arg Asn Thr Ala Glu Thr Leu Thr Tyr His
2900 2905 2910
Pro Pro His Thr Pro Leu Ile Thr Thr Asn Pro Thr Asp His Asp Pro
2915 2920 2925
Thr Thr Pro His Tyr Trp Val Arg Gln Ala Arg Glu Thr Val His Tyr
2930 2935 2940
Ala His Thr Thr Gln Gln Leu His Thr His Gly Val Thr Ala Tyr Leu
2945 2950 2955 2960
Glu Leu Gly Pro Asp His Thr Leu Thr Ala Leu Thr His His Asn Leu
2965 2970 2975
Pro Asp His Thr Pro Leu Ala Val Pro Leu Leu His Pro Asp Gln Ser
2980 2985 2990
Glu Thr His Thr Thr His Thr Ala Leu Ala His Leu His Thr His Gly
2995 3000 3005
His Pro Thr Thr Trp His His His His Thr Pro Thr His Tyr His Pro
3010 3015 3020
Asn Leu Pro Thr Tyr Pro Phe Gln His His His Tyr Trp Leu Asn Thr
3025 3030 3035 3040
Thr Thr Ala Thr Gly Asp Met Ser Ala Ala Gly Leu Glu Pro Ala Arg
3045 3050 3055
His Pro Leu Leu Gly Ala Ala Val Gly Leu Ala Asp Gly Glu Gly Leu
3060 3065 3070
Leu Phe Thr Gly Arg Ile Ser Leu Arg Thr His Pro Trp Leu Ala Asp
3075 3080 3085
His Ala Val Gly Gly Ala Val Leu Leu Pro Gly Thr Ala Phe Leu Glu
3090 3095 3100
Leu Ala Leu Gln Ala Ala Ala His Ala Asp Cys Arg Arg Val Glu Glu
3105 3110 3115 3120
Leu Thr Leu His Thr Pro Leu Val Val Pro Asp Ser Ala Gly Val Val
3125 3130 3135
Leu Gln Val Thr Val Ala Ala Pro Asn Glu Ala Gly Asn Arg Ala Val
3140 3145 3150
Asp Ile Tyr Ser Arg Ile Asp Val Gly Gly Leu Thr Ala Asp Ser Ala
3155 3160 3165
Gly Glu Pro Trp Thr Arg His Ala Ala Gly Tyr Leu Ala Asp Lys Pro
3170 3175 3180
Asp Pro Asp Cys Gly Asp Ser Ala Asp Gly Val Met Pro Ala Gly Ala
3185 3190 3195 3200
Trp Pro Pro Pro Gly Ala Val Ala Val Asp Leu Glu Gly Leu Tyr Glu
3205 3210 3215
Gln Leu Ala Glu Gly Gly Phe His Tyr Gly Ala Ala Phe Arg Cys Leu
3220 3225 3230
Asp Ala Ala Trp Gln Arg Gly Asp Glu Val Phe Ala Thr Ala Tyr Met
3235 3240 3245
Ser Glu Asp Gln Leu Gly Asp Thr Ala Ala Ala Arg Phe Ala Leu His
3250 3255 3260
Pro Ala Leu Leu Asp Ser Ala Leu His Thr Ile Pro Leu Leu Pro Ser
3265 3270 3275 3280
Leu Arg Gly Gln Gln Asp Ser Gly Leu Pro Phe Thr Trp Thr Gly Val
3285 3290 3295
Thr Leu Arg Ala Ser Gly Ala Thr Ala Leu Arg Val Arg Leu Arg Pro
3300 3305 3310
Asp Gly His Gly Pro Gly Ala Val Ser Val Asp Val Ser Asp Glu Ala
3315 3320 3325
Gly Glu Pro Val Ala Ser Val Arg Ser Leu Ala Leu Arg Pro Val Thr
3330 3335 3340
Arg Ala Glu Leu His Thr Ala Glu Leu Arg Thr Ala Ala Pro Val Ala
3345 3350 3355 3360
Pro His Gly Ser Leu Phe Glu Val Arg Trp Glu Pro Val Pro Gln Pro
3365 3370 3375
Ser Ala Ala Glu Glu Ala Ala Pro Trp Val Met Ile Gly Thr Gly Pro
3380 3385 3390
Thr Leu Arg Pro Val Glu Asp Phe Val Thr Pro Pro Glu Arg Thr Tyr
3395 3400 3405
Ala Asp Leu Ala Ala Leu Cys Val Ala Ile Ala Asp Asp Ala Pro Val
3410 3415 3420
Pro Arg Thr Val Val Ala Trp Ser Pro Ala Gly Ser Glu Asp Glu Ser
3425 3430 3435 3440
Ser Glu Ala Leu Arg Gln Ala Thr His His Met Leu Gly Leu Leu Gln
3445 3450 3455
Gln Trp Leu Ala Asp Ser Arg Phe Ala Asp Ser Arg Leu Val Ile Leu
3460 3465 3470
Thr Arg Ala Ala Val Ala Thr Ala Pro Asp Glu Glu Val Glu Asp Leu
3475 3480 3485
Ala Gly Ala Ala Ala Arg Gly Leu Ile Arg Ser Ala Gln Ser Glu His
3490 3495 3500
Pro Asp Arg Phe Val Leu Leu Asp Leu Asp Asp Arg Pro Ala Asp Ala
3505 3510 3515 3520
Lys Asp His Asp Arg Met Leu Ser Met Ala Leu Ala Cys Gly Glu Pro
3525 3530 3535
Glu Val Ala Val Arg Asp Gly Ala Leu Arg Thr Pro Arg Leu Ser Pro
3540 3545 3550
Leu Ala Gly Thr Ala Thr Glu Ala Met Asp Glu His Pro Trp Asp Gln
3555 3560 3565
Asp Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ser Leu Gly Ala Met
3570 3575 3580
Leu Ala Arg His Leu Val Ala Thr His Gly Val Arg His Leu Met Leu
3585 3590 3595 3600
Ile Ser Arg Arg Gly Leu Asp Ala Pro Gly Ala Arg Arg Leu Gly Val
3605 3610 3615
Glu Leu Ala Glu Leu Gly Ala Gln Val Thr Ile Thr Ala Cys Asp Ala
3620 3625 3630
Ala Asp Gln Arg Gln Leu Ala Asn Val Leu Ser Glu Ile Ser Val Asp
3635 3640 3645
His Pro Leu Thr Ala Val Val His Ala Ala Gly Val Leu Asp Asp Gly
3650 3655 3660
Val Ile Thr Ser Leu Thr Pro Glu Gly Leu Thr His Val Leu Arg Ala
3665 3670 3675 3680
Lys Val Asp Ser Ala Leu Asn Leu His Gln Leu Thr Arg Asp Leu Pro
3685 3690 3695
Leu Ser Ala Phe Val Leu Phe Ser Ser Leu Ala Gly Val Met Gly Ser
3700 3705 3710
Ala Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu
3715 3720 3725
Ala Ser His Arg Arg Ala Ala Arg Leu Pro Ala Val Ser Leu Ala Trp
3730 3735 3740
Gly Val Trp Glu Gln Thr Glu Gly Met Thr Gly Gln Leu Glu Ala Thr
3745 3750 3755 3760
Asp His Ala Arg Leu Arg Arg Ser Gly Leu Arg Pro Leu Ala Ile Ser
3765 3770 3775
Glu Gly Leu Glu Leu Phe Asp Lys Ala Leu Ser Cys Gly His Ala Leu
3780 3785 3790
Val Val Pro Ala Ala Leu Ser Thr Arg Glu Leu Gln Thr Ser Gly Ser
3795 3800 3805
Val Pro Pro Phe Leu Arg His Leu Thr Gly Val Ala Pro Ala Arg Pro
3810 3815 3820
Ser Arg Thr Arg Asp Ala Ser Ala Gly Glu Pro Thr Ser Leu Arg Arg
3825 3830 3835 3840
Arg Leu Thr Gly Leu Gly Pro Glu Glu Arg Leu Arg Glu Val Leu Arg
3845 3850 3855
Leu Val Arg Ser Arg Ala Ala Ala Val Leu Gly His Gly Thr Ala Glu
3860 3865 3870
Ser Val Pro Ala Asp Ser Ala Phe Arg Asp Leu Gly Phe Asp Ser Leu
3875 3880 3885
Ala Ala Val Asp Leu Arg Asn Arg Leu Gln Gln Ala Thr Gly Leu Arg
3890 3895 3900
Leu Pro Ala Gly Leu Ile Phe Asp Arg Pro Arg Pro Asp Val Leu Ala
3905 3910 3915 3920
Arg Phe Leu Cys Asp Glu Leu Ala Gly Ala Gly Gly Thr Ser Ala Ala
3925 3930 3935
Thr Ala Ala Pro Pro Val Ala Ala Val Gly Gly Ala Ala Gly Glu Pro
3940 3945 3950
Val Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly Val Arg Ser
3955 3960 3965
Ala Glu Gly Leu Trp Asp Leu Val Ala Ser Gly Met Asp Ala Val Gly
3970 3975 3980
Asp Phe Pro Ala Asp Arg Gly Trp Glu Val Glu Arg Leu Tyr Asp Pro
3985 3990 3995 4000
Asp Pro Asp Arg Thr Gly Thr Ser Tyr Thr Arg Gln Gly Gly Phe Leu
4005 4010 4015
Tyr Asp Ala Gly Glu Phe Asp Ala Ala Phe Phe Gly Ile Gly Pro Arg
4020 4025 4030
Glu Ala Val Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Ile Ser
4035 4040 4045
Trp Glu Ala Leu Glu Arg Ala Gly Ile Asp Pro Ala Ser Leu Arg Gly
4050 4055 4060
Ser Ser Thr Gly Val Phe Ala Gly Val Met Tyr His Asp Tyr Gly Thr
4065 4070 4075 4080
Arg Leu Arg Glu Ile Pro Glu Gly Tyr Glu Gly Tyr Ile Gly Asn Gly
4085 4090 4095
Asn Ala Gly Ser Val Ala Ser Gly Arg Val Ala Tyr Thr Phe Gly Leu
4100 4105 4110
Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val
4115 4120 4125
Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ser Gly Glu Cys Ser Met
4130 4135 4140
Ala Leu Ala Gly Gly Val Thr Val Met Ser Thr Pro Thr Thr Phe Val
4145 4150 4155 4160
Glu Phe Ser Arg Gln Arg Gly Leu Ala Pro Asp Gly Arg Cys Lys Ser
4165 4170 4175
Phe Gly Ala Gly Ala Asp Gly Thr Gly Trp Ala Glu Gly Ala Gly Met
4180 4185 4190
Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Asn Gly His Arg Val
4195 4200 4205
Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn
4210 4215 4220
Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Glu Arg Val Ile Arg Gln
4225 4230 4235 4240
Ala Trp Ala Asn Ala Gly Val Ala Ala Met Asp Ile Asp Ala Val Glu
4245 4250 4255
Gly His Gly Thr Gly Thr Thr Leu Gly Asp Pro Ile Glu Ala Gln Ala
4260 4265 4270
Leu Leu Gly Thr Tyr Gly Gln Gly Arg Ser Ala Asp Arg Pro Leu Trp
4275 4280 4285
Leu Gly Ser Ile Lys Ser Asn Val Gly His Thr Gln Ala Ala Ala Gly
4290 4295 4300
Val Gly Gly Val Ile Lys Met Val Met Ala Met Arg His Gly Leu Leu
4305 4310 4315 4320
Pro Gln Thr Leu His Ala Glu Glu Pro Ser Pro His Val Asp Trp Ser
4325 4330 4335
Gly Gly Thr Val Arg Leu Leu Thr Glu Ser Val Ala Trp Pro Glu Gln
4340 4345 4350
Gly Arg Met Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr
4355 4360 4365
Asn Ala His Val Ile Leu Glu Gln Ala Pro Pro Ala Ala Glu Thr His
4370 4375 4380
Glu Pro Ala Glu Pro Asn Thr Ala Pro Gly Pro Leu Pro Trp Ala Ile
4385 4390 4395 4400
Ser Ala Lys Ser Pro Gln Ala Leu Arg Ala Gln Ala Arg Gln Leu His
4405 4410 4415
Thr Tyr Leu Thr Asn Ala Pro Glu Ala Asn Pro Ala Asp Val Gly His
4420 4425 4430
Thr Leu Ala Thr Gly Arg Ala Ser Phe Glu His Arg Ala Val Val Ile
4435 4440 4445
Gly Ser Asp Arg Ala Glu Phe Leu Gly Gly Leu Asp Ala Leu Ala Ala
4450 4455 4460
Asp Glu Ala His Thr Ala Val Val Thr Gly Ile Ala Arg Lys Ala Gly
4465 4470 4475 4480
Asp Gln Gly Lys Val Val Phe Val Phe Pro Gly Gln Gly Gly Gln Trp
4485 4490 4495
Ala Gly Met Gly Leu Arg Leu Leu Lys Thr Ser Pro Val Phe Ala Gln
4500 4505 4510
Ser Ile Gln Ala Cys Glu Gln Ala Leu Ala Pro His Thr Asp Trp Thr
4515 4520 4525
Leu Thr Asp Ile Leu His Arg Pro His Thr Asp Pro Leu Trp Gln Arg
4530 4535 4540
Ala Asp Val Ile Gln Pro Val Leu Phe Ala Leu Met Thr Ser Leu Ala
4545 4550 4555 4560
Ala Leu Trp Gln Ser His Gly Leu Asn Pro Asp Ala Val Ile Gly His
4565 4570 4575
Ser Gln Gly Glu Ile Thr Ala Ala His Ile Ser Gly Ala Leu Ser Leu
4580 4585 4590
Glu Asp Ala Ala Lys Thr Val Ala Leu Arg Ser Arg Ala Leu Gln Thr
4595 4600 4605
Leu Arg Gly Ser Gly Gly Met Ala Ser Val Pro Leu Pro Ala Asp Gln
4610 4615 4620
Val Thr Gly Leu Leu Gln Thr Met Trp Pro Asp Arg Leu Trp Val Ala
4625 4630 4635 4640
Ala Val Asn Ala Pro Thr Ala Thr Val Ile Ser Gly Asn Ala Glu Ala
4645 4650 4655
Leu Thr Gln Ala Leu Glu His Tyr Arg Asp Gln Gly Val Asp Ala Lys
4660 4665 4670
Arg Ile Pro Val Asp Tyr Ala Ser His Cys Pro His Ile Gln Ala Val
4675 4680 4685
Glu Gln Glu Leu Ser Arg Leu Leu Arg Gly Ile Thr Pro Arg Ala Ala
4690 4695 4700
Thr Thr Pro Phe Tyr Ser Thr Thr Asp Asn Gln Trp Thr Asp Thr Thr
4705 4710 4715 4720
Thr Leu Asn Ala His Tyr Trp Tyr Arg Asn Leu Arg Gln Pro Val His
4725 4730 4735
Leu Ala Asp Ala Ile Thr Asn Leu Thr His Gln Gly His His Thr Phe
4740 4745 4750
Ile Glu Ile Ser Pro His Pro Thr Leu Thr Pro Ala Ile Gln Glu Thr
4755 4760 4765
Thr Asp Thr Thr His Thr Pro Thr Thr Val Ile Ser Thr Leu Arg Arg
4770 4775 4780
Asn His Asn Asp Thr His Gln Ile Leu His Ala Leu Ala His Ala His
4785 4790 4795 4800
Thr Thr Gly His Pro Ile Asn Trp His Thr Thr His Gln His His Thr
4805 4810 4815
Pro Thr Pro Gln His Ile Asp Leu Pro Thr Tyr Pro Phe Gln His His
4820 4825 4830
His Tyr Trp Leu Asn Thr Pro Thr Gln Thr Gly Asp Ala Ala Ala Val
4835 4840 4845
Gly Leu Asp Pro Ala His His Pro Leu Leu Gly Ala Ala Val Ala Val
4850 4855 4860
Ala Glu Gly Glu Gly Tyr Leu Leu Thr Gly Arg Leu Ala Leu Ser Thr
4865 4870 4875 4880
His Pro Trp Leu Ala Asp His Thr Ile Ala Gly Ala Val Val Leu Pro
4885 4890 4895
Gly Thr Ala Leu Leu Glu Ile Ala Leu Gln Ala Gly His Arg Val Asp
4900 4905 4910
Cys Trp Arg Ile Glu Glu Leu Thr Leu Gln Ser Pro Leu Phe Ile Pro
4915 4920 4925
Glu Glu Gly Ala Val Gln Val Gln Ala Trp Val Ala Ala Pro Asp Glu
4930 4935 4940
Asn Gly Cys Arg Ser Leu Thr Val Ser Ser Arg Arg Glu Gly Thr Tyr
4945 4950 4955 4960
Glu Asp Ala Thr Trp Val Arg His Ala Thr Gly Arg Val Gly Pro Ala
4965 4970 4975
Pro Ala Asp Gln Asp Glu Ala Ile Ala Arg Leu Thr Asp Pro Gln Gly
4980 4985 4990
Asp Gly Ala Ala Ala Ala Val Trp Pro Pro Gln Gly Ala Val Ala Phe
4995 5000 5005
Thr Ala Asp Asp Leu Glu Gly Leu Tyr Asp Gly Tyr Ala Ala Arg Gly
5010 5015 5020
Phe Glu Tyr Gly Pro Val Phe Arg Gly Leu Arg Ala Ala Trp Arg Arg
5025 5030 5035 5040
Gly Glu Asp Ile Phe Ala Glu Val Arg Leu Pro Asp Thr Ala Asp Gly
5045 5050 5055
Asp Ala Ser Gln Phe Ser Val His Pro Ala Leu Leu Asp Ala Ala Leu
5060 5065 5070
His Ala Ala Ala Phe Arg Pro Ala Asp Lys Leu Pro His Gly Ala Leu
5075 5080 5085
Pro Phe Ser Phe Ser Gly Val Arg Leu His Gly Pro Gly Ala Ser Thr
5090 5095 5100
Leu Arg Val Arg Leu Thr Pro Asp Gly Gln Ala Arg Asp Thr His Ala
5105 5110 5115 5120
Trp Ser Val Ala Val Val Asp Gly Glu Gly Arg Pro Val Ala Ser Ile
5125 5130 5135
Ala Ser Leu Ala Val Arg Pro Val Ser Thr Gln Glu Leu Leu Ala Ala
5140 5145 5150
Ser Gly Thr Ala Arg Arg Asp Ser Leu Phe Ala Val Glu Trp Val Thr
5155 5160 5165
Ala Leu Ala Pro Thr Ser Ser Ser Val Pro Gln Arg Leu Ala Thr Val
5170 5175 5180
Gly Pro Ser Asp Arg Leu Pro Ser Ala Asp Ala Tyr Ala Asn Leu Ala
5185 5190 5195 5200
Asp Leu Ala Ala Ala Val Leu Glu Ala Gly Ala Pro Ala Pro Asp Ala
5205 5210 5215
Val Val Val Asp Cys Gly Arg Arg Asp Ala Arg Ala Thr Ala Val Pro
5220 5225 5230
Glu Asp Val Arg Thr Leu Thr Arg Arg Ile Leu Gly Leu Leu Gln Glu
5235 5240 5245
Trp Leu Ala Asp Glu Arg Pro Ala Ser Ser Arg Met Val Val Leu Thr
5250 5255 5260
Arg Gly Ala Val Ala Thr Thr Pro Gly Glu Asp Val Ala Asp Leu Ala
5265 5270 5275 5280
Gly Ala Ala Val Cys Gly Met Val Arg Ser Ala Gln Ser Glu His Pro
5285 5290 5295
Gly Arg Phe Val Leu Leu Asp Leu Asp Pro Asp Pro Asp Leu Asp Gly
5300 5305 5310
Gly Glu Val Pro Pro Thr Val Val Pro Ala Ala Leu Ala Cys Gly Glu
5315 5320 5325
Pro Gln Ile Ala Val Arg Ala Asn Arg His Leu Val Pro Arg Leu Thr
5330 5335 5340
Arg Val Pro Ala Ser Val Pro Val Pro Gly Arg Val Pro Val Pro Ala
5345 5350 5355 5360
Ala Glu Ala Ala Asp Pro Asp Thr Thr Pro Thr Ala Phe Asp Pro Asp
5365 5370 5375
Gly Thr Val Val Ile Thr Gly Gly Thr Gly Thr Leu Gly Ala Met Leu
5380 5385 5390
Ala Arg His Leu Val Ser Arg His Gly Val Arg His Leu Leu Leu Ala
5395 5400 5405
Ser Arg Arg Gly Pro Asp Ala Pro Gly Ala Thr Glu Leu Arg Ala Glu
5410 5415 5420
Leu Ala Glu Leu Gly Ala Glu Val Thr Val Arg Ala Cys Asp Thr Gly
5425 5430 5435 5440
Asp Arg Gly Ala Leu Ala Asp Leu Ile Ala Gly Ile Pro Thr Gly His
5445 5450 5455
Pro Leu Thr Gly Val Val His Ala Ala Gly Val Leu Asp Asp Ala Thr
5460 5465 5470
Val Ala Ser Leu Thr Pro Arg His Leu Asp Thr Ala Leu Thr Pro Lys
5475 5480 5485
Ala Asp Ala Ala Phe His Leu His Glu Leu Thr Arg His Ala Arg Pro
5490 5495 5500
Arg Ala Phe Val Leu Phe Ser Ser Ala Ala Gly Val Leu Gly Ala Ala
5505 5510 5515 5520
Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala
5525 5530 5535
Glu His Arg Arg Ala Gln Gly Leu Pro Ala Leu Ser Leu Ala Trp Gly
5540 5545 5550
Leu Trp Glu Gln Gly Ser Gly Met Thr Gly His Leu Asp Arg Thr Asp
5555 5560 5565
Arg Ala Arg Ile Asn Arg Ser Gly Leu Ala Pro Leu Ala Thr Glu Asp
5570 5575 5580
Ala Leu Ala Leu Phe Asp Ala Ala Leu Ala Gly Asp Arg Pro Phe Leu
5585 5590 5595 5600
Val Pro Ala Arg Leu Asp Leu Arg Gly Ser Ser Ala Ala Glu Thr Pro
5605 5610 5615
Ala Pro Leu Phe Ser Arg Ile Ala Pro Ala Arg Thr Thr Arg Gly Arg
5620 5625 5630
Ser Pro Gly Ala Glu Gly Ala Ala Asp Leu Arg Thr Arg Leu Ala Ala
5635 5640 5645
Gln Asp Ala Ala Glu Gln Arg Asp Thr Leu Leu Thr Ile Val Arg Thr
5650 5655 5660
His Thr Ala Ala Val Leu Gly His Asp Thr Ala Ala Ala Val Arg Pro
5665 5670 5675 5680
Asp Gly Ala Phe Arg Glu Leu Gly Phe Asp Ser Leu Ala Ala Val Glu
5685 5690 5695
Leu Arg Asn Arg Leu Gln Thr Thr Thr Ala Leu Thr Leu Pro Ala Thr
5700 5705 5710
Thr Val Phe Asp His Pro Thr Pro Ala Ala Leu Ala Asp His Leu Arg
5715 5720 5725
Thr Gln Leu Cys Gln Asp Ala Gln Ser Ser Ala Ala Ala Thr Ala Met
5730 5735 5740
Ala Ala Met Ala Glu Leu Ala Arg Leu Glu Ser Ala Val Ser Asp Ser
5745 5750 5755 5760
Val Ala Leu Asp Asp Asp Thr Arg Ser Gly Leu Ala Glu Arg Leu Arg
5765 5770 5775
Ser Leu Ala Arg Lys Met Ser Ser Gly Arg Val Val Asp His Asp Gly
5780 5785 5790
Gly Gly Ala Ala Asp Leu Asp Leu Gln Ser Val Thr Asp Asp Glu Met
5795 5800 5805
Phe Glu Leu Ile Asp Lys Glu Val Ser Arg Asp
5810 5815
<210> 75
<211> 17469
<212> DNA
<213> Artificial Sequence
<220>
<223> meiA3 gene of Streptomyces nanchangensis
<400> 75
ttggagatac cgatggccgc tggccacgac aaggtgatcg aggcgctgcg ggcgtccctc 60
aagaccaacg agcggcagag ggaacagatc caccggctca ctacggcggc gcgggaaccc 120
atcgccatca tcggcatggc ctgccgctat cccggcggag tgggatcgcc ggaggacctg 180
tgggagctgg tggccgccgg ccgtgacgcc atcggcacct tccccgagga ccggggctgg 240
gacgcggcgc ggctgtacga ccccgatccg gagcgggccg gcacctcgta cacccagcat 300
ggcggattcc tttaccaggc aggggagttc gaccccggtt tcttcgggat cagcccgcgc 360
gaggcgctgg cgatggaccc gcagcagcgg ctgctgctgg agatctcctg ggaggcgttc 420
gagcgggccg ggatcgaccc ggcctcggtg cgcggcagcc gcaccggggt cttcgcgggc 480
gtcatgtacc acgactacgg ctcccggctg cacaccgtcc ccgaaggctt cgagggctac 540
gtcggcaacg gcagcggcgg cggcgtggcg tccggccggg tcgcctacac cctcggcctc 600
gaaggcccgg ccgtgaccgt ggacaccgcc tgctcctcct cactggtcgc cctgcacctg 660
gcctgccagg cgctgcgggc cggcgagtgc tcactcgccc tcgcgggcgg ggtgacggtg 720
atgtccaccc ccagcctgtt cgtcgagtac tcccggcagc gcgcgctcgc ggcggatggc 780
cggtgcaagg cgtacggggc gggggcggac ggcaccggct gggcagaagg cgccgggatg 840
ttgctggtgg aacggctcac ggacgcacag cggctcggcc accgggtgct ggcagtggtc 900
cggggcagcg cggtcaacca ggacggcgcg agcaacggcc tcaccgcccc caacggcccc 960
gcgcagcaac gggtcatccg gcaggcactg gcgagcgccg gggtgtcggc gtccgaggtc 1020
gacgccgtgg aggggcatgg gacggggacg cggctgggcg atccgatcga ggcgcaggcg 1080
ttgctggcga cctacggtca gcagcggccc gcggaccggc cgctgtggct cgggtcgatg 1140
aagtccaacg tcggccatgc gcaggcggcc gccggcgtgg gcgggatcat caagatggtg 1200
atggccatgc ggagcgggac gctgccgcgc accctgcacg cggacgagcc gtcgccgcac 1260
atcgactggg actcgggcgc ggtgcggctg ctgaccgagc cggtcgcctg gccggagcgc 1320
gaccggccgc gccgcgccgc ggtgtcctcc ttcggggtca gcggcaccaa cgcccatgtg 1380
atcctcgagg ccgcgtcgca gacggcgccg cacacggaat ccgcgtcgca gacggaaacc 1440
gacgacgctc ccgcgccgca cggcgcgccg ggccatgccg tggcggggcc gctgccctgg 1500
cccctgtcgg gcgcgacggc cgaggcgctg cgggcccagg ccagggagct gcgtcgcttc 1560
gtggcggccg atgagctgct gcgccccgcc gacgtcgggc acaccctggt cttgggccgc 1620
tcggacctcg cacaccgcgc agtcgtcctc ggctccgacc gggaaaccct gctgcgcggt 1680
ctggacactc tgacagggga ggggccggac ggcggctcgg tcgtacgggg cgtggcggcg 1740
gcaggggccg gtgcgggcgt ggtgttcgtc ttcccgggac agggcggcca gtgggccggc 1800
atggggctgc ggctgctgga gacctcgtcg ttcttcgccg agcggatggc ggagtgcgag 1860
gcggccttgg caccgtatgt cgactggtcg ctgctcgacg tgctgcgccg ggaccccggg 1920
gacccggtgt gggagcgggc cgatgtcgtc cagccgatgc tgttctcggt gatggtgtcg 1980
ctggcgcagc tgtggcgctc gtatggcgtc gaaccggacg ccgtactcgg ccactcccag 2040
ggcgagatcg ccgccgccca catctgcggc gcgctgaccc tggacgacgc cgcgaaggtt 2100
gtcgccctgc gcagccgggc cctgcagacc ctgcgcggtt cgggcggcat ggcctccgta 2160
ccactgacgg cggacgaggt cgccgggctg ctgcggaccg catggccgga ccggctgtgg 2220
gtggccgccg tcaacgcccc cacggccacg gtgatctccg gcgacgcgga ctctctggcg 2280
gaggcgctgg aacactaccg ggaccagggc gtcgacgcga agcgggtccc ggtcgactac 2340
gcctcccact gcccgcatat cgaggccgtg gagcaggagc tgctgagcct gttgcggggg 2400
atcgctccaa gggccgccga cattcccttc tactccactg tggacaacca gtgggccgac 2460
accatgggac tcgacgcccg gtactggtac cgcaatctgc gccggcccgt acgcttcgcc 2520
gaagcgctcc gcgctctcgg tgccgccgag taccggtcgt atgtcgaggt cggcccgcac 2580
cccaccctca cccccgccat cgaggacacc actgaggccg ccggcgccgc ggccacggtt 2640
gtcggctccc tgcgccgcgg cgaggacgac gcccaccgca tcctcacctc gctggcccgg 2700
gctcatattc atggcctgcc cgtggcgtgg gaccgccact accgggcgct cgcccccgag 2760
gcgaaccatg tcgacctgcc cacctacgcc ttccagcgcc gccgctactg gctggacgcc 2820
ccggcgacca ccggggacgt gacggccgcg gggctggccc cggtcggaca cccactgctc 2880
ggcgcggcgg tcggactcgc cgagggcgac ggatatctgc tcaccggccg gctcgccccg 2940
cacacccacc cctggctcac cgaccacgcg gtcgccggca ccgtcctgct gccgggcacc 3000
gcatacgtgg aactggccgt gcacgtcggc gaacacctcg gctgcccccg gctggaggag 3060
ctcaccctgc acgccccgct cgtcctcccc gacacgggcg gtgtggcgct ccaggtggcc 3120
gtcggcgcac cggacgagac cggccgccgc gcactgagcg tctacgcaca gcgcgacgac 3180
gaccccacgt gggaaggggc ggcccggggc gcgtggacac ggcatgcgac cggcacactg 3240
gcggccgagg ccgcgaccga tggcatcaac ggtgccgacg gtgccgggcc cctggcgggg 3300
gcgtggcctc cgccgggcgc ggagcccctg gacatcagcg gcctctacga cacgctggcc 3360
gccgcggact tcggctacgg cccggccttc caggggctgc gcgccgtctg gcggcacggc 3420
gaggagacct acgccgaggt gcggctcccc gaccaggtgg ccgccgacgc cccacgcttc 3480
tgcctccacc ccgcgttgct cgacgccgcg ctccacccgc tggcactcga cagcggccga 3540
agcgaggaga atccagcggg acatggcctg ctgccattcg cctggcgcgg cgtcagcctg 3600
cgctccccgg gcacaccgac gctgcgcgta cggctgcggc cgcagggccc ggactcgatt 3660
gccgtcgacg tggccgacga gacgggcgcg tcggtggtct cggccgaatc gctcacgctg 3720
cgaccggtgg ccctggagga cctgcgggtc ctcggcggcc aggcgaacga ccccctctat 3780
gccctggagt ggaccgccgc gcccgagccc ctgacaacag ccctcgggcg gtgcgccgtg 3840
cttggccacg ccacccccgg atgggccgcc gcgttggaga cggcggcagc ggagcccgta 3900
cggcggtacc cggaccttgc cggactggta gcggccctgg acgccggcga tccgcctccg 3960
gacctggtgt tcgtgggctg ccctccggct gccgccgggc ccgacgacac gacggtcgcc 4020
gacgttcaca ccacccgtac ccgtgtccgt acccgacaag cgctggagct gcttcaaggc 4080
tggctcggcg aagcgcggct ggccggcgcg cggctggtgc tggtcacccg cggcgcggtg 4140
gccaccgggc cggcgggggg agggatggac ctggcgggcg cggcgatctg cggactggtg 4200
cgatccgcac aggccgagga gcccgatcgc atcctcctgg tggacttgga cacggccgag 4260
gagtcgtggg cggcgctgcc acgggcggtc gcgctgggcg aaccgcagat ggccatccgg 4320
gccggccagc cgcacatggc ccggctggtg cgagccgaca ccgagaggga cgccctgctc 4380
acgccgccac gggggagcgg cggctggcgg ctcgactgcg ccgatgcggg cacgctccag 4440
gggttggcgc cggtggcgtc ctcggccgac cacgacccgc tgggcccgca gcaggtacgg 4500
atcgaggtgc gtgcggccgg gctgaacttc cgcgatgtcc tggtggccct ggggatggtc 4560
cctgggcagc aggggctggg cagcgagggc gccggggtgg tgctcgaagc cgggcctgaa 4620
gtggccgacc tggcgcccgg agaccgggtg atgggcgtgt tcgcggacgc gttcggcccg 4680
ttcgcgatcg ccgaccgggc cacagtgatc cgcgtccccg agcactggac cttcgcccag 4740
gccgccgccg tccccgtcgt gttcgccacc gcctactacg ggctggtgga cctggcagga 4800
ctgcgcccgg gcgagtcggt gctggtgcac gccgcggccg gcggagtggg actggccgcc 4860
gtccaactgg cccgccacct gggcgctgag gtctacgcca cggcgagccc cggcaaatgg 4920
gacaccctac gcgcccacgg catccccccg gagcgcatcg cctcgtcccg caccctcgac 4980
ttcgagagcc ggttcaccgg ccggaacatc gacgtcgtcc tcaactccct ggcccatgag 5040
tacgtcgacg cctcgctgcg cctggtgtcc ggcgacagcg gccggttcct cgagatgggc 5100
aagaccgacc tccgtgaccc ggaggaggtg gcggaggcgt accccggtgt cgcctaccgg 5160
gcgtacgacc tgatggaggc cggacccgag cgcatcgggg agatcctgcg caccgtgctg 5220
cggctgttcg acgagggcgt gctcaccccg ctgccgctca cctgctggga catccggcag 5280
gccagggatg ccttccgcca actccagcag ggccgcaccg tcggaaagaa tgtgctcacg 5340
ctggaccgca cccccgaccc cgacggcacc gtcctcatca ccggtggcac cggcaccctc 5400
ggcgccgcgc tcgcccgcca tctcgccgcc accggccgag cacggcatct gctgctgatc 5460
agccgccgtg gcctcgatgc gccaggcgct cccgaactca tcgctgagat cgacgagttg 5520
ggcgcggcga cgaccgtcgc cacctgcgac gtcggcgacc gtgccgcgct cgccgaactg 5580
ctcgggcgga tccccgccga gcacccgctg accgccgtcg tccacgccgc gggcacactc 5640
gacgacgcca cgctcggctc cctcaccgcg cgccacctcg acaccgttct gcccgcgaag 5700
gccgatgccg cctggcatct gcacgagctg acctgccggc tggatctggc cgcgttcgtg 5760
ctgttctcgt ccgccgcggg cgtcctgggc tcgccggggc agggcaacta cgccgccgcc 5820
aatgcctttc tcgacgcgct cgccttccag cgacgggcga tgggactccc cgccgtgtcc 5880
ctggcatggg gactgtggga ggaggccagc gggatgaccg gccacctcga ccagaccgac 5940
cgcacccgca tggcccgcgt cggcctccgg ccactggcca cgaacgaggc cctggcgctg 6000
ttcgacaacg ctctcgtcga tggcccaccg ctgctgctcc cggcccgtat cgacaccaag 6060
gcgctacggg gcaccaccgc accgcccctg ttccagagcc tcgtacgtcc caccaccggc 6120
caccggccac gccccgcgac acccgacggc cgctcctccc tccgagcccg gctcgccggg 6180
ctcgaccctg ccgcacagca cgaggtcctg ctcaccctcg tccgcggcca cgccgccacg 6240
gtcctcggcc acccgagccc cgacgccatc gcccccgagg cggccttccg tgacctcggc 6300
ttcgactccc tcaccgccgt agagctccgc aaccgcctca aggaggcaac cggtctgcgg 6360
ctccccgcca ccctcgtctt cgaccacccc actcctgccg ctctcgccca gcacctgcgg 6420
gacggcctca tcggcggcgc cgatgcggcc accttggctt cggctcctgc tccgagcgag 6480
gtggcgacgg tggcggatga ggccatcgcg atcatcggca tggcctgccg gtatccgggg 6540
ggcgtgcggt cggccgaagg gctgtgggat ctggtcgcct ccggcaccga cgccatgagc 6600
ggattcccca ccgaccgcgg ctgggacctc gaccgcctct acgcccccca ggaccaggac 6660
cggccgggca ccacatacac ccgccacggg ggcttcctcc acgacgcggg caagttcgac 6720
gcgggattct tcggcatcgg cccacgtgag gcgctggcga tggatccaca gcagcggctg 6780
ctgctggaga cctcctggga ggttttcgaa cacgcgggaa tcgacccctc gtcggtacgg 6840
cggagccgga ccggagtctt cgccggtgtg atgccgacgg actacggccc ccggctgcaa 6900
gacaccgtgg ccgaggtcga gggctatgtc ctcaccggaa actccggcag cgtcgcctcg 6960
ggccgtatcg cctacacctt cggtctggaa ggccccgcgg tgtcggtgga cacggcgtgt 7020
tcgtcgtctc tggtggcgtt gcatctggcg tgtcaggcgc tgcgtgcggg ggagtgctcc 7080
atggcgctgg ccggcggggt gacggtgatg gcgacgcctg gtgccttcgt ggagtttgcg 7140
cggcagcggg ggttgtcggt ggatgggcgg tgcaaggcgt ttggggtggg tgcggatggt 7200
acggggtggg cggagggggt ggggatgctg ttggtggagc ggttgtctga tgcgcggcgg 7260
ttggggcatc gggtgttggc ggtggtgcgg ggttctgcgg tgaatcagga cggggcgagc 7320
aatggtttga cggcgccgaa tggtccgtcg cagcagcggg tgatccggca ggcgttggcc 7380
agtgcgcggg ttggtggggc ggatgtggat gtggtggagg ggcacggtac ggggacgcgg 7440
ctgggtgatc cgatcgaggc gcaggcgttg ctggcgacct acggtcagga gcggtcgggg 7500
gatgaaccgt tgtggttggg gtcggtgaag tcgaatatcg ggcatgcgca ggctgcggcg 7560
ggtgttgcgg gtgtcatcaa gatggtgatg gcgatgcggt gtggggtgtt gccgcggacg 7620
ttgcatgtgc aggagccgtc gccgcatgtg gactggtcct cgggtggggt gcggctgctg 7680
acggaggcgg tgccgtggcc ggagacgggt cgtgcgcggc gtgcgggggt gtcgtcgttc 7740
ggggtcagcg gcaccaacgc gcacatcatc ctcgaacagg caccgccgga ggagcacgac 7800
gatccggcgg acgtttcgtc cgggtcgttt ccgtggatgg tgtcggccaa gtccgaacag 7860
gcactacagg cacaggcagc gcagctgcgc gcgtatctgg cggcacgtcc cggggtgggg 7920
ctggctgatg tcgggtatgc gctggccgcc ggccgtaccg ccttcgacca ccgtgccgtg 7980
ctcctgggcc cggaccgcga agccttcctc gaagggctgg gggctctggg ggccggtgag 8040
gaacacgccg ggctcgtacg gggcgtggcg acgggtgcgg ggaagctggc gttcgtgtgt 8100
tccgggcagg gcacgcagcg ccctcgtatg gggcacgagc tgtaccgcgc cttcccgctg 8160
ttcgccgcag ccatggacga agcctgcgca tacctggacc cgcatctcga ccggcctctg 8220
cgggatgtcg tgttcgccga gccggactcc ggtacggccc ggctgctgca gcagacgcgc 8280
tatgcccagc ccgcgctgtt cgccctccag gtcgccctgc atcgcctggt caccgaacac 8340
tacggcctca cgccccacta ctacgcgggc cattccctgg gggagatcac cgcggcccac 8400
ctcgccggga tcctgaccct ctgcgacgcg gcgcgtctgg tcaccacccg cgcccgcctg 8460
atgcagtctc tccccgccac cggcgcgatg accaccctcc aagcagaccc cgacgaactc 8520
cacgaacacc tcgcacgatg cgagggacgg gtgtcgctcg cggccgtgaa cgcgcctggg 8580
tccgtggtca tcagcggtga ccgccacgac gtagacgcca cggccgaaaa cttccgcgcc 8640
atggggcgca agaccacccc gttgaaggtc agcggcgcct tccactcaca ccacatcgac 8700
ccactcctcg acgaactccg cgccaccgcc gaaaccctca cctaccaccc accccacacc 8760
cccctcatca cgaccgacct gaccgaccag gaccccacca cacctggcta ttgggtccgg 8820
caaacacgcg agaccgtcca ctacgcccac accacccaac aactccacac ccacggcgtc 8880
accgcctacc tcgaactcgg ccccgacacc acactcacca ccctcaccca ccacaacctc 8940
ccccaccaca cccccctagc catccccctc ctccaccccg accaacccga aacccacacc 9000
acccacaccg ccctcgccca cctccacacc cacggccacc ccaccacctg gcaccaccac 9060
cacaccccca cccaccacca cccaaacctc cccacctacc ccttccaaca ccaccactac 9120
tggctcaaca ccaccactgc caccggtgat atgtcggcgg caggccttga gccggcgcgg 9180
catcccctgt tgggcgcggc ggtcgagttg gccgatggtg aggggttgct gttcactggg 9240
cggatttcac tccgtacgca tccctggttg gccgaccacg ccgtcggcgg cgccgtgttg 9300
ctccccggta cggcctttct cgaactcgcc ctcgaagccg ccgcccatgt cgactgccat 9360
cggatcgagg agcttacgct ccacaccccg ctcgtcgtac cggagagcgg cggcgtagtg 9420
ctgcaggtga ccgtggccgg gccgaacgaa gcaggaaacc gggcggtgga tatctactcg 9480
cgaatcgatg tcggcggcct caccgccgat tcggtgggcg agccgtggac gcgccatgcc 9540
gccgggtacc ttgccgacaa gcctggccca gactgcggtg actcggcgga tggtgtcatg 9600
cctgcgggcg catggccgcc gccgggtgcg gtcgccgtgg atctggagga actgtacgag 9660
cagctggccg aggggggttt ccactacggt gcggccttcc gttgcctgga cgccgcctgg 9720
caacgcggcg acgaggtctt cgcgactgtg catatgtcag agaatcagct gggcgacacg 9780
gccgcggctc ggttcgcgct gcaccccgcg ctgctggatt ccgcactgca caccattcca 9840
ctcctcccct ccctgcaggg acaacaggac agcgggctgc cgttcacgtg ggcaggagtc 9900
accctgcgcg catccggggc cacggccctg cgcgtccggc tgaggccgga tggccatggc 9960
ccgggggcgg tgtccgtcga cgtgtccgac gaggcgggtg agcccgtagc atcagttcgg 10020
tcgttggccc tgcggccggt gaccagggtc gagttgcata cggccgagtt gcgcacagcc 10080
gccccagttg ccccccatag ctcgctcttc gaggtgcgat gggaacccgt cccccagccc 10140
tcagcggccg aagaagccga tccatgggtg atgatcggga ccggaccgac gctgcgcccg 10200
gacgaggact tcgccactcc gccggagcgg acgtacgccg acctggccgc gctgtgcgcg 10260
gcagtcgccg atggcgcgcc cgttccccgg acggtcgtgg cctggtccca ggccgggagc 10320
gaagacgagt cgagtgaggc gctgcgccac gccacacacc acatgctggg cctactgcag 10380
cagtggttgg cggacagccg gttcgtcgac agtcgcctgg tgatcctcac ccgagccgcg 10440
gtggccactg cgccggagga ggaggtaaaa gacctggcgg gagcggcgac gcggggtctg 10500
atccgctccg cccagtcgga gcaccccgac cgattcgtcc tgctcgacct ggacgaccgt 10560
cccgctgacg cgaaagacca cgaccgaatg ctgtcggtgg ccctggcctg cggggaaccg 10620
gaagtggccg tacgcgatgg agccctgcgc acaccccggc tgagcccgct tgccggcacc 10680
gccaccgagg ccatggacga gcatccctgg gatccggacg gcaccgtact catcaccggc 10740
ggcaccggca gcctcggcgc catgctcgcc cgccacttgg tggccaccca tggcgtacgg 10800
catctgctgc tgatcagccg acgtggcctc gacgccccgg gggccaggcg acaggggaac 10860
gaactcgtcg agctcggagc gcagttgacc atcgccgcgt gcgatgccgc agaccaaagg 10920
caacttgcaa acgcattgtc ggagatctcc gtcgaccatc cgctgaccgc tgtggtgcat 10980
gcggcaggcg tactggacga cggggtgatc acatccctca caccggagga cctgacccat 11040
gtcctgcggg ccaaggtcga ttcggcgctc aatctccacc agctcacacg cgacctgccg 11100
ctgtccgcgt ttgtgctctt ctcctcgctg gccggggtga tgggttcggc agggcagggc 11160
aactacgccg ccgccaacgc cgccctggac gcgctggcga gtcaccgcag ggccactcgg 11220
ctgccggcgg tgtccctggc ctggggagtt tgggagcaga ccgagggcat gaccgggcag 11280
ttggaggcca cgggccacgc gaggctccgc cgctcgggcc tgaggccgct ggccaccagc 11340
gagggcctgg agctcttcga caaggccttg agctgtggac acgccctggt ggtgcccgcc 11400
gcactcagca cgaaggagct tcagacatcc ggatccgtcc caccattcct gcgccacgtg 11460
acgggcgtcg ctccggcccg gccgtcccgg acccgcgacg cctcggccgg tgagccgacc 11520
cccctgcggc ggcggttgac cggcctcggg ccggaagagc ggctacgcga ggtgctgcgg 11580
ctggtgcgct cccgggcggc tgcggtgctg gggcacggca cggccgaagc ggtcccggcg 11640
gactcggcgt tccgcgacct ggggttcgac tccctcgccg cggtggacct gcggaaccgg 11700
ttgcagcagg ccaccgggct gcgcctgccg gccggcttga tcttcgaccg gccgcgtccg 11760
gacgtactcg cccgtttcct gtgtgacgag ttggccggtg tcggcggtac gtcggcggcc 11820
acggccgccc cacccgttgc ggccgtcggc ggggcagccg gcgagccggt ggccatcgtc 11880
ggcatggcat gccggtttcc gggaggtgtg cggtcggccg agggcctgtg ggatctggtc 11940
gcctccggta tggacgcggt gggtgacttc cccacagacc gaggctggga ggtggaacgg 12000
ctctacgacc ccgacccgga ccgaaccggc acctcctata cccggcaagg cgggttcctc 12060
tacgacgcgg gtgagttcga cgcggcgttc ttcgggattg gcccgcgtga ggcggtggcg 12120
atggatccac agcagcggct gctgctggag atttcctggg aggcgctgga acgggcggga 12180
atcgacccgg cgtcgctgcg ggggagttcg actggagtgt tcgctggggt gatgtaccac 12240
gactacggca cccgcttgcg cgagatccca gagggctacg agggctatat cggcaatgga 12300
aacgcgggca gcgtcgcttc gggacgtgtc tcctacactt tcggcctgga ggggccggcg 12360
gtcaccgtgg acacggcgtg ttcgtcgtcc ctggtcgccc tgcatctggc ctgccaggcg 12420
ctgcggtcag gggagtgctc catggcgctg gcgggcgggg tcaccgtcat gtccaccccc 12480
accacttttg tcgagttctc gcgccaacgg ggactggccc cggacgggcg gtgcaagtcc 12540
ttcggggccg gcgcggacgg aacgggctgg gcggagggcg cggggatgct cctggtggag 12600
cggctttcgg acgcccggcg caacggccac cgggtcctgg cggtggtacg ggggagcgcg 12660
gtcaaccagg acggggcgag caatgggctg acggcgccga acggcccgtc gcaagagcgg 12720
gtgatccgcc aggcgtgggc aaatgcgggt gtggccgcga tggacatcga cgcggtggag 12780
ggacacggca cggggacgac gctcggtgac cccattgagg cccaggcgct gctggggacg 12840
tatggacagg gacggtcggc cgatcggccg ttgtggttgg gatcgatcaa gtccaacgtc 12900
ggacacaccc aggccgccgc gggggtgggc ggcgtcatca agatggtgat ggccatgcgc 12960
cacgggctgc tcccgcagac cctgcacgcc gaggagccct cacctcatgt ggactggtcg 13020
ggcgggacgg tgcggttgct gaccgagccg gtggcctggc ctgagcgggg gcggatgcgc 13080
cgcgcaggcg tctcctcttt cggtgtcagc ggtaccaacg cccacgtcat cttggaacaa 13140
gcaccaccta acgcggagac ccacgaaccg gcagagcccc acaccgcgcc aggcccactg 13200
ccctggacga tctccgcgaa gagcccgcaa gcgctacgtg cccaggcgcg tcagttgcac 13260
acgtacctga ccaacacccc cgaggcgaac cccgccgacg tcggccacac cctcgcgatg 13320
ggccgcgcct ctttcgagca tcgtgcggtg gttatcggct ccgatcgagg ggagtttctg 13380
ggtggtctgg atgctgtggc ggcagatgag gcccactctg ctgtggtcac gggtatcgcg 13440
aggaaggccg gtgacctggg gaaggtggtg ttcgtcttcc ccgggcaggg tggtcagtgg 13500
gccgggatgg gactgcggct gctcaagacc tcgcccgtct tcgcgcaatc catccaggcc 13560
tgcgaacaag ccctcgcccc ccacaccgac tggaccctga ccgacatcct gcaccgcccc 13620
cacaccgacc ccctgtggca gcgcgccgac gtcatccagc ccgccctctt cgccctcatg 13680
acctccctca ccaccctctg gcaatcccac ggcctcaacc ccgacgccgt catcggccac 13740
tcccaaggcg aaatcaccgc cgcccacgcc tgcggagcac tgagcctgga agacgccgcg 13800
aaaatcgtcg ccctccgcag ccagaccctg caaaccctcc aaggctcagg cggcatggcc 13860
tccgtaccac tgcccgcaga ccaggtcacc gcactgctgc acaccatgtg gcccgaccag 13920
ctatgggtcg ccgccatcaa cgcccccacc accacagtca tctccggcga cacacaagcc 13980
ctcacacaag cgctgaacca ctaccgggac caagacatcg acgcgaaacg catcccggtc 14040
gactacgcct cccactgccc ccacatccag gccgtccaac acgaactctc agacctgttg 14100
caggacatca ccccacgggc cgcgaccacc cccttctact ccaccaccga caaccaatgg 14160
accgacacca ccaccctcaa cgcccactac tggtaccgaa acctccgcca acccgtccac 14220
ctcaccaacg ccatcaccaa cctcacccac caaggccacc acacctacat cgaaatcagc 14280
ccccacccca ccctcacccc cgccatccag gaaaccaccc acaccaccca cacccccacc 14340
accgtcatca gcacactccg ccgcaaccac aacgacaccc accaactcct ccacgccctc 14400
gcccacgccc acaccaccgg ccaccccatc aactggcacc ccacccacca acaccacacc 14460
ccaacccccc aacacaccga cctccccacc taccccttcc aacaccaacg ctactggctc 14520
aacaccccca cccaaacagg agacgcagca gccatcggcc tggacccggc acatcacccg 14580
ctgctcggcg cggcggtcgc agtggccgag ggggagggct atctgctcac cggtcggctc 14640
gccctgtcca cccacccctg gcttgccgat cacaccatcg cgggcgcggt cgtccttccc 14700
ggaactgccc ttcttgagat cgcccttcag gcgggccatc gtgtggactg ccatcgcatc 14760
gaagaactca ccctccaatc gccgctgttc atcccggaag agggagcagt acaggtgcag 14820
gcatgggtgg cggcgccgga tgagaacggg taccgaagcc tgacggtgtc ctcccgacgt 14880
gagggtacgt acgaggacgc cacgtgggtg cgccatgcca cgggccgggt cggtcccgca 14940
ccggccgacc aggatgatgc catcgcgcgg ctcaccgacc cacaaggcga cggagcggcg 15000
gcggtctggc caccgcaggg cgctgtcgcg ttcacagcag acgatctgga gggcctgtac 15060
gacgggtacg cggcgcgggg attcgagtac ggcccggtgt tccgaggact gcgggcggcc 15120
tggcgacgtg gcgaggacat cttcgccgag gtgcgccttc ccgacacggc ggacggcgac 15180
gcctcccagt tctccgtaca ccccgccctg ctggacgccg ccctgcacgc cgccgccttc 15240
cgcccggccg acgaactccc gcacggggct ctgcccttct ccttcagcgg ggtgaggctg 15300
cacgggcccg gagcgtcgac cctgcgggtg cgcctcaccc cggatggcca ggcgcgggac 15360
acgcacgcat ggtcggtcgc ggtggtcgac ggcgaggggc ggccggtggc ctcgatcgcg 15420
tcgctcgcgg tccgcccggt gtcgacgcag gagttgctgg cggcctccgg tacggcgcgg 15480
cgggactcgc tcttcgcggt cgagtgggtg accgccccgg cgccgacctc gtcgtccgct 15540
ccgcgacgcc tggccacggt ggggcccagc gaccgcctcc cctcggcaga cgcgtacgcg 15600
aacctcgccg acctggccgc cgcagtgctg gaggcggagg ccccggcgcc cgatgcggtc 15660
gtggtcgact gcggccgccg cgacgcgcgc gccacggccg tggcggagga cgtacggacc 15720
ctcacccggc gcatcctggg tctgctgcag gagtggctgg cggacgagag gccggcctcg 15780
agccggatgg tcgtactgac ccgtggtgcg gtggccacca caccggggga ggacgtggcg 15840
gacctggcgg gcgcggcggt gtgcggcatg gtgcggtccg cgcagtcgga acatcccggc 15900
cggttcgtcc tgctggacct cgaccccgac ccggacctcg acggcgggga agtgccaccg 15960
accgtcgtac cggcggctct cgcctgtggt gagccgcaga tcgcggtgcg tgcgaaccgg 16020
cacctggtgc cccggctgac ccgcgttccg gtgtccgtcc ccgtccccgg gcctgttccc 16080
gttcccgccg ccgaggcagc cgaccaggac accacgccca cggcgttcga ccccgacggc 16140
accgtactga tcaccggcgg caccggcacc ctcggcgcgg tgctcgcgcg ccatctggtc 16200
agccgtcacg gcgtacggca cctgctgctg gcatcgcgac gcgggcccga cgcacccggc 16260
gccaccgagc tgcgggcgga actggccgag ctcggggccg aggcgacggt gcgcgcttgt 16320
gacaccggtg accgaggcgc gctggcggat ctcatcgcgg ggattcccac cggccaccct 16380
ttgaccggtg tggtccacgc cgcgggcgtc ctggatgacg ccaccgtcgc ctccctcacc 16440
ccccgacacc tggacaccgc gctgacaccc aaggccgacg ccgccttcca tctgcacgag 16500
ctcacccgcc acgcccggcc gcgcgccttc gtcctgttct cctcggccgc cggtgtcctc 16560
ggcgcagccg ggcagggcaa ctacgctgcc gccaacgcct tcctcgacgc cctcgccgaa 16620
caccgcaggg cgcagggcct gccggccttg tcgctcgcgt ggggcttgtg ggagcagggc 16680
agcggcatga ccgggcatct cgaccgcacc gaccgggccc gcatcaaccg ctccggactc 16740
gcccccctcg ccaccgagga cgctctcgcg ctcttcgacg ccgccctcgc cggcgatcgg 16800
ccgttcctgg tgcccgcccg gctggaccta cggggttcaa gcgccgccga gaccccggcg 16860
ccgctgttct ccaggatcgc cccggctcgt acgacccggg gccgtacccc cggcgctgag 16920
ggcgccgctg accttcgtac ccgtctcgcg gcccaggatg ccaccgagca gcgcgacacg 16980
cttctcacga tcgtccgcac ccacaccgcc gccgtcctgg ggcatgacac ggctgccgcc 17040
gtgcggccgg acgcggcctt ccgtgagctg ggtttcgact ccctcgccgc cgtggaactc 17100
cgtaaccgcc ttcaaacgac caccgccctc accctgcccg cgaccaccgt tttcgaccac 17160
cccacgcccg ctgccctcgc cgatcatctg cgtactcagc tctgccagga cgctccgtcc 17220
ccggcggcgg ccacggccat ggcggcgatg gcggagctgg ccaggctgga gtccgccgtc 17280
tccgattcgg cggcgctcga cgacgacacg cgcagcggcc tcgcggagcg cctgcggtcc 17340
ctcgcccgca agatgagcag tggccgtgtc gtcgaccaca acggcggcgg cgctgcgggc 17400
ctggatctcc agtcggccac ggacgatgag atgttcgagc tgatcgacaa ggaggtcagc 17460
cgagactga 17469
<210> 76
<211> 5822
<212> PRT
<213> Artificial Sequence
<220>
<223> meiA3 protein of Streptomyces nanchangensis
<400> 76
Met Glu Ile Pro Met Ala Ala Gly His Asp Lys Val Ile Glu Ala Leu
1 5 10 15
Arg Ala Ser Leu Lys Thr Asn Glu Arg Gln Arg Glu Gln Ile His Arg
20 25 30
Leu Thr Thr Ala Ala Arg Glu Pro Ile Ala Ile Ile Gly Met Ala Cys
35 40 45
Arg Tyr Pro Gly Gly Val Gly Ser Pro Glu Asp Leu Trp Glu Leu Val
50 55 60
Ala Ala Gly Arg Asp Ala Ile Gly Thr Phe Pro Glu Asp Arg Gly Trp
65 70 75 80
Asp Ala Ala Arg Leu Tyr Asp Pro Asp Pro Glu Arg Ala Gly Thr Ser
85 90 95
Tyr Thr Gln His Gly Gly Phe Leu Tyr Gln Ala Gly Glu Phe Asp Pro
100 105 110
Gly Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln
115 120 125
Gln Arg Leu Leu Leu Glu Ile Ser Trp Glu Ala Phe Glu Arg Ala Gly
130 135 140
Ile Asp Pro Ala Ser Val Arg Gly Ser Arg Thr Gly Val Phe Ala Gly
145 150 155 160
Val Met Tyr His Asp Tyr Gly Ser Arg Leu His Thr Val Pro Glu Gly
165 170 175
Phe Glu Gly Tyr Val Gly Asn Gly Ser Gly Gly Gly Val Ala Ser Gly
180 185 190
Arg Val Ala Tyr Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Val Asp
195 200 205
Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala
210 215 220
Leu Arg Ala Gly Glu Cys Ser Leu Ala Leu Ala Gly Gly Val Thr Val
225 230 235 240
Met Ser Thr Pro Ser Leu Phe Val Glu Tyr Ser Arg Gln Arg Ala Leu
245 250 255
Ala Ala Asp Gly Arg Cys Lys Ala Tyr Gly Ala Gly Ala Asp Gly Thr
260 265 270
Gly Trp Ala Glu Gly Ala Gly Met Leu Leu Val Glu Arg Leu Thr Asp
275 280 285
Ala Gln Arg Leu Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala
290 295 300
Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro
305 310 315 320
Ala Gln Gln Arg Val Ile Arg Gln Ala Leu Ala Ser Ala Gly Val Ser
325 330 335
Ala Ser Glu Val Asp Ala Val Glu Gly His Gly Thr Gly Thr Arg Leu
340 345 350
Gly Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Gln
355 360 365
Arg Pro Ala Asp Arg Pro Leu Trp Leu Gly Ser Met Lys Ser Asn Val
370 375 380
Gly His Ala Gln Ala Ala Ala Gly Val Gly Gly Ile Ile Lys Met Val
385 390 395 400
Met Ala Met Arg Ser Gly Thr Leu Pro Arg Thr Leu His Ala Asp Glu
405 410 415
Pro Ser Pro His Ile Asp Trp Asp Ser Gly Ala Val Arg Leu Leu Thr
420 425 430
Glu Pro Val Ala Trp Pro Glu Arg Asp Arg Pro Arg Arg Ala Ala Val
435 440 445
Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Ala
450 455 460
Ala Ser Gln Thr Ala Pro His Thr Glu Ser Ala Ser Gln Thr Glu Thr
465 470 475 480
Asp Asp Ala Pro Ala Pro His Gly Ala Pro Gly His Ala Val Ala Gly
485 490 495
Pro Leu Pro Trp Pro Leu Ser Gly Ala Thr Ala Glu Ala Leu Arg Ala
500 505 510
Gln Ala Arg Glu Leu Arg Arg Phe Val Ala Ala Asp Glu Leu Leu Arg
515 520 525
Pro Ala Asp Val Gly His Thr Leu Val Leu Gly Arg Ser Asp Leu Ala
530 535 540
His Arg Ala Val Val Leu Gly Ser Asp Arg Glu Thr Leu Leu Arg Gly
545 550 555 560
Leu Asp Thr Leu Thr Gly Glu Gly Pro Asp Gly Gly Ser Val Val Arg
565 570 575
Gly Val Ala Ala Ala Gly Ala Gly Ala Gly Val Val Phe Val Phe Pro
580 585 590
Gly Gln Gly Gly Gln Trp Ala Gly Met Gly Leu Arg Leu Leu Glu Thr
595 600 605
Ser Ser Phe Phe Ala Glu Arg Met Ala Glu Cys Glu Ala Ala Leu Ala
610 615 620
Pro Tyr Val Asp Trp Ser Leu Leu Asp Val Leu Arg Arg Asp Pro Gly
625 630 635 640
Asp Pro Val Trp Glu Arg Ala Asp Val Val Gln Pro Met Leu Phe Ser
645 650 655
Val Met Val Ser Leu Ala Gln Leu Trp Arg Ser Tyr Gly Val Glu Pro
660 665 670
Asp Ala Val Leu Gly His Ser Gln Gly Glu Ile Ala Ala Ala His Ile
675 680 685
Cys Gly Ala Leu Thr Leu Asp Asp Ala Ala Lys Val Val Ala Leu Arg
690 695 700
Ser Arg Ala Leu Gln Thr Leu Arg Gly Ser Gly Gly Met Ala Ser Val
705 710 715 720
Pro Leu Thr Ala Asp Glu Val Ala Gly Leu Leu Arg Thr Ala Trp Pro
725 730 735
Asp Arg Leu Trp Val Ala Ala Val Asn Ala Pro Thr Ala Thr Val Ile
740 745 750
Ser Gly Asp Ala Asp Ser Leu Ala Glu Ala Leu Glu His Tyr Arg Asp
755 760 765
Gln Gly Val Asp Ala Lys Arg Val Pro Val Asp Tyr Ala Ser His Cys
770 775 780
Pro His Ile Glu Ala Val Glu Gln Glu Leu Leu Ser Leu Leu Arg Gly
785 790 795 800
Ile Ala Pro Arg Ala Ala Asp Ile Pro Phe Tyr Ser Thr Val Asp Asn
805 810 815
Gln Trp Ala Asp Thr Met Gly Leu Asp Ala Arg Tyr Trp Tyr Arg Asn
820 825 830
Leu Arg Arg Pro Val Arg Phe Ala Glu Ala Leu Arg Ala Leu Gly Ala
835 840 845
Ala Glu Tyr Arg Ser Tyr Val Glu Val Gly Pro His Pro Thr Leu Thr
850 855 860
Pro Ala Ile Glu Asp Thr Thr Glu Ala Ala Gly Ala Ala Ala Thr Val
865 870 875 880
Val Gly Ser Leu Arg Arg Gly Glu Asp Asp Ala His Arg Ile Leu Thr
885 890 895
Ser Leu Ala Arg Ala His Ile His Gly Leu Pro Val Ala Trp Asp Arg
900 905 910
His Tyr Arg Ala Leu Ala Pro Glu Ala Asn His Val Asp Leu Pro Thr
915 920 925
Tyr Ala Phe Gln Arg Arg Arg Tyr Trp Leu Asp Ala Pro Ala Thr Thr
930 935 940
Gly Asp Val Thr Ala Ala Gly Leu Ala Pro Val Gly His Pro Leu Leu
945 950 955 960
Gly Ala Ala Val Gly Leu Ala Glu Gly Asp Gly Tyr Leu Leu Thr Gly
965 970 975
Arg Leu Ala Pro His Thr His Pro Trp Leu Thr Asp His Ala Val Ala
980 985 990
Gly Thr Val Leu Leu Pro Gly Thr Ala Tyr Val Glu Leu Ala Val His
995 1000 1005
Val Gly Glu His Leu Gly Cys Pro Arg Leu Glu Glu Leu Thr Leu His
1010 1015 1020
Ala Pro Leu Val Leu Pro Asp Thr Gly Gly Val Ala Leu Gln Val Ala
1025 1030 1035 1040
Val Gly Ala Pro Asp Glu Thr Gly Arg Arg Ala Leu Ser Val Tyr Ala
1045 1050 1055
Gln Arg Asp Asp Asp Pro Thr Trp Glu Gly Ala Ala Arg Gly Ala Trp
1060 1065 1070
Thr Arg His Ala Thr Gly Thr Leu Ala Ala Glu Ala Ala Thr Asp Gly
1075 1080 1085
Ile Asn Gly Ala Asp Gly Ala Gly Pro Leu Ala Gly Ala Trp Pro Pro
1090 1095 1100
Pro Gly Ala Glu Pro Leu Asp Ile Ser Gly Leu Tyr Asp Thr Leu Ala
1105 1110 1115 1120
Ala Ala Asp Phe Gly Tyr Gly Pro Ala Phe Gln Gly Leu Arg Ala Val
1125 1130 1135
Trp Arg His Gly Glu Glu Thr Tyr Ala Glu Val Arg Leu Pro Asp Gln
1140 1145 1150
Val Ala Ala Asp Ala Pro Arg Phe Cys Leu His Pro Ala Leu Leu Asp
1155 1160 1165
Ala Ala Leu His Pro Leu Ala Leu Asp Ser Gly Arg Ser Glu Glu Asn
1170 1175 1180
Pro Ala Gly His Gly Leu Leu Pro Phe Ala Trp Arg Gly Val Ser Leu
1185 1190 1195 1200
Arg Ser Pro Gly Thr Pro Thr Leu Arg Val Arg Leu Arg Pro Gln Gly
1205 1210 1215
Pro Asp Ser Ile Ala Val Asp Val Ala Asp Glu Thr Gly Ala Ser Val
1220 1225 1230
Val Ser Ala Glu Ser Leu Thr Leu Arg Pro Val Ala Leu Glu Asp Leu
1235 1240 1245
Arg Val Leu Gly Gly Gln Ala Asn Asp Pro Leu Tyr Ala Leu Glu Trp
1250 1255 1260
Thr Ala Ala Pro Glu Pro Leu Thr Thr Ala Leu Gly Arg Cys Ala Val
1265 1270 1275 1280
Leu Gly His Ala Thr Pro Gly Trp Ala Ala Ala Leu Glu Thr Ala Ala
1285 1290 1295
Ala Glu Pro Val Arg Arg Tyr Pro Asp Leu Ala Gly Leu Val Ala Ala
1300 1305 1310
Leu Asp Ala Gly Asp Pro Pro Pro Asp Leu Val Phe Val Gly Cys Pro
1315 1320 1325
Pro Ala Ala Ala Gly Pro Asp Asp Thr Thr Val Ala Asp Val His Thr
1330 1335 1340
Thr Arg Thr Arg Val Arg Thr Arg Gln Ala Leu Glu Leu Leu Gln Gly
1345 1350 1355 1360
Trp Leu Gly Glu Ala Arg Leu Ala Gly Ala Arg Leu Val Leu Val Thr
1365 1370 1375
Arg Gly Ala Val Ala Thr Gly Pro Ala Gly Gly Gly Met Asp Leu Ala
1380 1385 1390
Gly Ala Ala Ile Cys Gly Leu Val Arg Ser Ala Gln Ala Glu Glu Pro
1395 1400 1405
Asp Arg Ile Leu Leu Val Asp Leu Asp Thr Ala Glu Glu Ser Trp Ala
1410 1415 1420
Ala Leu Pro Arg Ala Val Ala Leu Gly Glu Pro Gln Met Ala Ile Arg
1425 1430 1435 1440
Ala Gly Gln Pro His Met Ala Arg Leu Val Arg Ala Asp Thr Glu Arg
1445 1450 1455
Asp Ala Leu Leu Thr Pro Pro Arg Gly Ser Gly Gly Trp Arg Leu Asp
1460 1465 1470
Cys Ala Asp Ala Gly Thr Leu Gln Gly Leu Ala Pro Val Ala Ser Ser
1475 1480 1485
Ala Asp His Asp Pro Leu Gly Pro Gln Gln Val Arg Ile Glu Val Arg
1490 1495 1500
Ala Ala Gly Leu Asn Phe Arg Asp Val Leu Val Ala Leu Gly Met Val
1505 1510 1515 1520
Pro Gly Gln Gln Gly Leu Gly Ser Glu Gly Ala Gly Val Val Leu Glu
1525 1530 1535
Ala Gly Pro Glu Val Ala Asp Leu Ala Pro Gly Asp Arg Val Met Gly
1540 1545 1550
Val Phe Ala Asp Ala Phe Gly Pro Phe Ala Ile Ala Asp Arg Ala Thr
1555 1560 1565
Val Ile Arg Val Pro Glu His Trp Thr Phe Ala Gln Ala Ala Ala Val
1570 1575 1580
Pro Val Val Phe Ala Thr Ala Tyr Tyr Gly Leu Val Asp Leu Ala Gly
1585 1590 1595 1600
Leu Arg Pro Gly Glu Ser Val Leu Val His Ala Ala Ala Gly Gly Val
1605 1610 1615
Gly Leu Ala Ala Val Gln Leu Ala Arg His Leu Gly Ala Glu Val Tyr
1620 1625 1630
Ala Thr Ala Ser Pro Gly Lys Trp Asp Thr Leu Arg Ala His Gly Ile
1635 1640 1645
Pro Pro Glu Arg Ile Ala Ser Ser Arg Thr Leu Asp Phe Glu Ser Arg
1650 1655 1660
Phe Thr Gly Arg Asn Ile Asp Val Val Leu Asn Ser Leu Ala His Glu
1665 1670 1675 1680
Tyr Val Asp Ala Ser Leu Arg Leu Val Ser Gly Asp Ser Gly Arg Phe
1685 1690 1695
Leu Glu Met Gly Lys Thr Asp Leu Arg Asp Pro Glu Glu Val Ala Glu
1700 1705 1710
Ala Tyr Pro Gly Val Ala Tyr Arg Ala Tyr Asp Leu Met Glu Ala Gly
1715 1720 1725
Pro Glu Arg Ile Gly Glu Ile Leu Arg Thr Val Leu Arg Leu Phe Asp
1730 1735 1740
Glu Gly Val Leu Thr Pro Leu Pro Leu Thr Cys Trp Asp Ile Arg Gln
1745 1750 1755 1760
Ala Arg Asp Ala Phe Arg Gln Leu Gln Gln Gly Arg Thr Val Gly Lys
1765 1770 1775
Asn Val Leu Thr Leu Asp Arg Thr Pro Asp Pro Asp Gly Thr Val Leu
1780 1785 1790
Ile Thr Gly Gly Thr Gly Thr Leu Gly Ala Ala Leu Ala Arg His Leu
1795 1800 1805
Ala Ala Thr Gly Arg Ala Arg His Leu Leu Leu Ile Ser Arg Arg Gly
1810 1815 1820
Leu Asp Ala Pro Gly Ala Pro Glu Leu Ile Ala Glu Ile Asp Glu Leu
1825 1830 1835 1840
Gly Ala Ala Thr Thr Val Ala Thr Cys Asp Val Gly Asp Arg Ala Ala
1845 1850 1855
Leu Ala Glu Leu Leu Gly Arg Ile Pro Ala Glu His Pro Leu Thr Ala
1860 1865 1870
Val Val His Ala Ala Gly Thr Leu Asp Asp Ala Thr Leu Gly Ser Leu
1875 1880 1885
Thr Ala Arg His Leu Asp Thr Val Leu Pro Ala Lys Ala Asp Ala Ala
1890 1895 1900
Trp His Leu His Glu Leu Thr Cys Arg Leu Asp Leu Ala Ala Phe Val
1905 1910 1915 1920
Leu Phe Ser Ser Ala Ala Gly Val Leu Gly Ser Pro Gly Gln Gly Asn
1925 1930 1935
Tyr Ala Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala Phe Gln Arg Arg
1940 1945 1950
Ala Met Gly Leu Pro Ala Val Ser Leu Ala Trp Gly Leu Trp Glu Glu
1955 1960 1965
Ala Ser Gly Met Thr Gly His Leu Asp Gln Thr Asp Arg Thr Arg Met
1970 1975 1980
Ala Arg Val Gly Leu Arg Pro Leu Ala Thr Asn Glu Ala Leu Ala Leu
1985 1990 1995 2000
Phe Asp Asn Ala Leu Val Asp Gly Pro Pro Leu Leu Leu Pro Ala Arg
2005 2010 2015
Ile Asp Thr Lys Ala Leu Arg Gly Thr Thr Ala Pro Pro Leu Phe Gln
2020 2025 2030
Ser Leu Val Arg Pro Thr Thr Gly His Arg Pro Arg Pro Ala Thr Pro
2035 2040 2045
Asp Gly Arg Ser Ser Leu Arg Ala Arg Leu Ala Gly Leu Asp Pro Ala
2050 2055 2060
Ala Gln His Glu Val Leu Leu Thr Leu Val Arg Gly His Ala Ala Thr
2065 2070 2075 2080
Val Leu Gly His Pro Ser Pro Asp Ala Ile Ala Pro Glu Ala Ala Phe
2085 2090 2095
Arg Asp Leu Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Arg Asn Arg
2100 2105 2110
Leu Lys Glu Ala Thr Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp
2115 2120 2125
His Pro Thr Pro Ala Ala Leu Ala Gln His Leu Arg Asp Gly Leu Ile
2130 2135 2140
Gly Gly Ala Asp Ala Ala Thr Leu Ala Ser Ala Pro Ala Pro Ser Glu
2145 2150 2155 2160
Val Ala Thr Val Ala Asp Glu Ala Ile Ala Ile Ile Gly Met Ala Cys
2165 2170 2175
Arg Tyr Pro Gly Gly Val Arg Ser Ala Glu Gly Leu Trp Asp Leu Val
2180 2185 2190
Ala Ser Gly Thr Asp Ala Met Ser Gly Phe Pro Thr Asp Arg Gly Trp
2195 2200 2205
Asp Leu Asp Arg Leu Tyr Ala Pro Gln Asp Gln Asp Arg Pro Gly Thr
2210 2215 2220
Thr Tyr Thr Arg His Gly Gly Phe Leu His Asp Ala Gly Lys Phe Asp
2225 2230 2235 2240
Ala Gly Phe Phe Gly Ile Gly Pro Arg Glu Ala Leu Ala Met Asp Pro
2245 2250 2255
Gln Gln Arg Leu Leu Leu Glu Thr Ser Trp Glu Val Phe Glu His Ala
2260 2265 2270
Gly Ile Asp Pro Ser Ser Val Arg Arg Ser Arg Thr Gly Val Phe Ala
2275 2280 2285
Gly Val Met Pro Thr Asp Tyr Gly Pro Arg Leu Gln Asp Thr Val Ala
2290 2295 2300
Glu Val Glu Gly Tyr Val Leu Thr Gly Asn Ser Gly Ser Val Ala Ser
2305 2310 2315 2320
Gly Arg Ile Ala Tyr Thr Phe Gly Leu Glu Gly Pro Ala Val Ser Val
2325 2330 2335
Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln
2340 2345 2350
Ala Leu Arg Ala Gly Glu Cys Ser Met Ala Leu Ala Gly Gly Val Thr
2355 2360 2365
Val Met Ala Thr Pro Gly Ala Phe Val Glu Phe Ala Arg Gln Arg Gly
2370 2375 2380
Leu Ser Val Asp Gly Arg Cys Lys Ala Phe Gly Val Gly Ala Asp Gly
2385 2390 2395 2400
Thr Gly Trp Ala Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser
2405 2410 2415
Asp Ala Arg Arg Leu Gly His Arg Val Leu Ala Val Val Arg Gly Ser
2420 2425 2430
Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly
2435 2440 2445
Pro Ser Gln Gln Arg Val Ile Arg Gln Ala Leu Ala Ser Ala Arg Val
2450 2455 2460
Gly Gly Ala Asp Val Asp Val Val Glu Gly His Gly Thr Gly Thr Arg
2465 2470 2475 2480
Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln
2485 2490 2495
Glu Arg Ser Gly Asp Glu Pro Leu Trp Leu Gly Ser Val Lys Ser Asn
2500 2505 2510
Ile Gly His Ala Gln Ala Ala Ala Gly Val Ala Gly Val Ile Lys Met
2515 2520 2525
Val Met Ala Met Arg Cys Gly Val Leu Pro Arg Thr Leu His Val Gln
2530 2535 2540
Glu Pro Ser Pro His Val Asp Trp Ser Ser Gly Gly Val Arg Leu Leu
2545 2550 2555 2560
Thr Glu Ala Val Pro Trp Pro Glu Thr Gly Arg Ala Arg Arg Ala Gly
2565 2570 2575
Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Ile Ile Leu Glu
2580 2585 2590
Gln Ala Pro Pro Glu Glu His Asp Asp Pro Ala Asp Val Ser Ser Gly
2595 2600 2605
Ser Phe Pro Trp Met Val Ser Ala Lys Ser Glu Gln Ala Leu Gln Ala
2610 2615 2620
Gln Ala Ala Gln Leu Arg Ala Tyr Leu Ala Ala Arg Pro Gly Val Gly
2625 2630 2635 2640
Leu Ala Asp Val Gly Tyr Ala Leu Ala Ala Gly Arg Thr Ala Phe Asp
2645 2650 2655
His Arg Ala Val Leu Leu Gly Pro Asp Arg Glu Ala Phe Leu Glu Gly
2660 2665 2670
Leu Gly Ala Leu Gly Ala Gly Glu Glu His Ala Gly Leu Val Arg Gly
2675 2680 2685
Val Ala Thr Gly Ala Gly Lys Leu Ala Phe Val Cys Ser Gly Gln Gly
2690 2695 2700
Thr Gln Arg Pro Arg Met Gly His Glu Leu Tyr Arg Ala Phe Pro Leu
2705 2710 2715 2720
Phe Ala Ala Ala Met Asp Glu Ala Cys Ala Tyr Leu Asp Pro His Leu
2725 2730 2735
Asp Arg Pro Leu Arg Asp Val Val Phe Ala Glu Pro Asp Ser Gly Thr
2740 2745 2750
Ala Arg Leu Leu Gln Gln Thr Arg Tyr Ala Gln Pro Ala Leu Phe Ala
2755 2760 2765
Leu Gln Val Ala Leu His Arg Leu Val Thr Glu His Tyr Gly Leu Thr
2770 2775 2780
Pro His Tyr Tyr Ala Gly His Ser Leu Gly Glu Ile Thr Ala Ala His
2785 2790 2795 2800
Leu Ala Gly Ile Leu Thr Leu Cys Asp Ala Ala Arg Leu Val Thr Thr
2805 2810 2815
Arg Ala Arg Leu Met Gln Ser Leu Pro Ala Thr Gly Ala Met Thr Thr
2820 2825 2830
Leu Gln Ala Asp Pro Asp Glu Leu His Glu His Leu Ala Arg Cys Glu
2835 2840 2845
Gly Arg Val Ser Leu Ala Ala Val Asn Ala Pro Gly Ser Val Val Ile
2850 2855 2860
Ser Gly Asp Arg His Asp Val Asp Ala Thr Ala Glu Asn Phe Arg Ala
2865 2870 2875 2880
Met Gly Arg Lys Thr Thr Pro Leu Lys Val Ser Gly Ala Phe His Ser
2885 2890 2895
His His Ile Asp Pro Leu Leu Asp Glu Leu Arg Ala Thr Ala Glu Thr
2900 2905 2910
Leu Thr Tyr His Pro Pro His Thr Pro Leu Ile Thr Thr Asp Leu Thr
2915 2920 2925
Asp Gln Asp Pro Thr Thr Pro Gly Tyr Trp Val Arg Gln Thr Arg Glu
2930 2935 2940
Thr Val His Tyr Ala His Thr Thr Gln Gln Leu His Thr His Gly Val
2945 2950 2955 2960
Thr Ala Tyr Leu Glu Leu Gly Pro Asp Thr Thr Leu Thr Thr Leu Thr
2965 2970 2975
His His Asn Leu Pro His His Thr Pro Leu Ala Ile Pro Leu Leu His
2980 2985 2990
Pro Asp Gln Pro Glu Thr His Thr Thr His Thr Ala Leu Ala His Leu
2995 3000 3005
His Thr His Gly His Pro Thr Thr Trp His His His His Thr Pro Thr
3010 3015 3020
His His His Pro Asn Leu Pro Thr Tyr Pro Phe Gln His His His Tyr
3025 3030 3035 3040
Trp Leu Asn Thr Thr Thr Ala Thr Gly Asp Met Ser Ala Ala Gly Leu
3045 3050 3055
Glu Pro Ala Arg His Pro Leu Leu Gly Ala Ala Val Glu Leu Ala Asp
3060 3065 3070
Gly Glu Gly Leu Leu Phe Thr Gly Arg Ile Ser Leu Arg Thr His Pro
3075 3080 3085
Trp Leu Ala Asp His Ala Val Gly Gly Ala Val Leu Leu Pro Gly Thr
3090 3095 3100
Ala Phe Leu Glu Leu Ala Leu Glu Ala Ala Ala His Val Asp Cys His
3105 3110 3115 3120
Arg Ile Glu Glu Leu Thr Leu His Thr Pro Leu Val Val Pro Glu Ser
3125 3130 3135
Gly Gly Val Val Leu Gln Val Thr Val Ala Gly Pro Asn Glu Ala Gly
3140 3145 3150
Asn Arg Ala Val Asp Ile Tyr Ser Arg Ile Asp Val Gly Gly Leu Thr
3155 3160 3165
Ala Asp Ser Val Gly Glu Pro Trp Thr Arg His Ala Ala Gly Tyr Leu
3170 3175 3180
Ala Asp Lys Pro Gly Pro Asp Cys Gly Asp Ser Ala Asp Gly Val Met
3185 3190 3195 3200
Pro Ala Gly Ala Trp Pro Pro Pro Gly Ala Val Ala Val Asp Leu Glu
3205 3210 3215
Glu Leu Tyr Glu Gln Leu Ala Glu Gly Gly Phe His Tyr Gly Ala Ala
3220 3225 3230
Phe Arg Cys Leu Asp Ala Ala Trp Gln Arg Gly Asp Glu Val Phe Ala
3235 3240 3245
Thr Val His Met Ser Glu Asn Gln Leu Gly Asp Thr Ala Ala Ala Arg
3250 3255 3260
Phe Ala Leu His Pro Ala Leu Leu Asp Ser Ala Leu His Thr Ile Pro
3265 3270 3275 3280
Leu Leu Pro Ser Leu Gln Gly Gln Gln Asp Ser Gly Leu Pro Phe Thr
3285 3290 3295
Trp Ala Gly Val Thr Leu Arg Ala Ser Gly Ala Thr Ala Leu Arg Val
3300 3305 3310
Arg Leu Arg Pro Asp Gly His Gly Pro Gly Ala Val Ser Val Asp Val
3315 3320 3325
Ser Asp Glu Ala Gly Glu Pro Val Ala Ser Val Arg Ser Leu Ala Leu
3330 3335 3340
Arg Pro Val Thr Arg Val Glu Leu His Thr Ala Glu Leu Arg Thr Ala
3345 3350 3355 3360
Ala Pro Val Ala Pro His Ser Ser Leu Phe Glu Val Arg Trp Glu Pro
3365 3370 3375
Val Pro Gln Pro Ser Ala Ala Glu Glu Ala Asp Pro Trp Val Met Ile
3380 3385 3390
Gly Thr Gly Pro Thr Leu Arg Pro Asp Glu Asp Phe Ala Thr Pro Pro
3395 3400 3405
Glu Arg Thr Tyr Ala Asp Leu Ala Ala Leu Cys Ala Ala Val Ala Asp
3410 3415 3420
Gly Ala Pro Val Pro Arg Thr Val Val Ala Trp Ser Gln Ala Gly Ser
3425 3430 3435 3440
Glu Asp Glu Ser Ser Glu Ala Leu Arg His Ala Thr His His Met Leu
3445 3450 3455
Gly Leu Leu Gln Gln Trp Leu Ala Asp Ser Arg Phe Val Asp Ser Arg
3460 3465 3470
Leu Val Ile Leu Thr Arg Ala Ala Val Ala Thr Ala Pro Glu Glu Glu
3475 3480 3485
Val Lys Asp Leu Ala Gly Ala Ala Thr Arg Gly Leu Ile Arg Ser Ala
3490 3495 3500
Gln Ser Glu His Pro Asp Arg Phe Val Leu Leu Asp Leu Asp Asp Arg
3505 3510 3515 3520
Pro Ala Asp Ala Lys Asp His Asp Arg Met Leu Ser Val Ala Leu Ala
3525 3530 3535
Cys Gly Glu Pro Glu Val Ala Val Arg Asp Gly Ala Leu Arg Thr Pro
3540 3545 3550
Arg Leu Ser Pro Leu Ala Gly Thr Ala Thr Glu Ala Met Asp Glu His
3555 3560 3565
Pro Trp Asp Pro Asp Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ser
3570 3575 3580
Leu Gly Ala Met Leu Ala Arg His Leu Val Ala Thr His Gly Val Arg
3585 3590 3595 3600
His Leu Leu Leu Ile Ser Arg Arg Gly Leu Asp Ala Pro Gly Ala Arg
3605 3610 3615
Arg Gln Gly Asn Glu Leu Val Glu Leu Gly Ala Gln Leu Thr Ile Ala
3620 3625 3630
Ala Cys Asp Ala Ala Asp Gln Arg Gln Leu Ala Asn Ala Leu Ser Glu
3635 3640 3645
Ile Ser Val Asp His Pro Leu Thr Ala Val Val His Ala Ala Gly Val
3650 3655 3660
Leu Asp Asp Gly Val Ile Thr Ser Leu Thr Pro Glu Asp Leu Thr His
3665 3670 3675 3680
Val Leu Arg Ala Lys Val Asp Ser Ala Leu Asn Leu His Gln Leu Thr
3685 3690 3695
Arg Asp Leu Pro Leu Ser Ala Phe Val Leu Phe Ser Ser Leu Ala Gly
3700 3705 3710
Val Met Gly Ser Ala Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ala
3715 3720 3725
Leu Asp Ala Leu Ala Ser His Arg Arg Ala Thr Arg Leu Pro Ala Val
3730 3735 3740
Ser Leu Ala Trp Gly Val Trp Glu Gln Thr Glu Gly Met Thr Gly Gln
3745 3750 3755 3760
Leu Glu Ala Thr Gly His Ala Arg Leu Arg Arg Ser Gly Leu Arg Pro
3765 3770 3775
Leu Ala Thr Ser Glu Gly Leu Glu Leu Phe Asp Lys Ala Leu Ser Cys
3780 3785 3790
Gly His Ala Leu Val Val Pro Ala Ala Leu Ser Thr Lys Glu Leu Gln
3795 3800 3805
Thr Ser Gly Ser Val Pro Pro Phe Leu Arg His Val Thr Gly Val Ala
3810 3815 3820
Pro Ala Arg Pro Ser Arg Thr Arg Asp Ala Ser Ala Gly Glu Pro Thr
3825 3830 3835 3840
Pro Leu Arg Arg Arg Leu Thr Gly Leu Gly Pro Glu Glu Arg Leu Arg
3845 3850 3855
Glu Val Leu Arg Leu Val Arg Ser Arg Ala Ala Ala Val Leu Gly His
3860 3865 3870
Gly Thr Ala Glu Ala Val Pro Ala Asp Ser Ala Phe Arg Asp Leu Gly
3875 3880 3885
Phe Asp Ser Leu Ala Ala Val Asp Leu Arg Asn Arg Leu Gln Gln Ala
3890 3895 3900
Thr Gly Leu Arg Leu Pro Ala Gly Leu Ile Phe Asp Arg Pro Arg Pro
3905 3910 3915 3920
Asp Val Leu Ala Arg Phe Leu Cys Asp Glu Leu Ala Gly Val Gly Gly
3925 3930 3935
Thr Ser Ala Ala Thr Ala Ala Pro Pro Val Ala Ala Val Gly Gly Ala
3940 3945 3950
Ala Gly Glu Pro Val Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly
3955 3960 3965
Gly Val Arg Ser Ala Glu Gly Leu Trp Asp Leu Val Ala Ser Gly Met
3970 3975 3980
Asp Ala Val Gly Asp Phe Pro Thr Asp Arg Gly Trp Glu Val Glu Arg
3985 3990 3995 4000
Leu Tyr Asp Pro Asp Pro Asp Arg Thr Gly Thr Ser Tyr Thr Arg Gln
4005 4010 4015
Gly Gly Phe Leu Tyr Asp Ala Gly Glu Phe Asp Ala Ala Phe Phe Gly
4020 4025 4030
Ile Gly Pro Arg Glu Ala Val Ala Met Asp Pro Gln Gln Arg Leu Leu
4035 4040 4045
Leu Glu Ile Ser Trp Glu Ala Leu Glu Arg Ala Gly Ile Asp Pro Ala
4050 4055 4060
Ser Leu Arg Gly Ser Ser Thr Gly Val Phe Ala Gly Val Met Tyr His
4065 4070 4075 4080
Asp Tyr Gly Thr Arg Leu Arg Glu Ile Pro Glu Gly Tyr Glu Gly Tyr
4085 4090 4095
Ile Gly Asn Gly Asn Ala Gly Ser Val Ala Ser Gly Arg Val Ser Tyr
4100 4105 4110
Thr Phe Gly Leu Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser
4115 4120 4125
Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ser Gly
4130 4135 4140
Glu Cys Ser Met Ala Leu Ala Gly Gly Val Thr Val Met Ser Thr Pro
4145 4150 4155 4160
Thr Thr Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Pro Asp Gly
4165 4170 4175
Arg Cys Lys Ser Phe Gly Ala Gly Ala Asp Gly Thr Gly Trp Ala Glu
4180 4185 4190
Gly Ala Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Asn
4195 4200 4205
Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp
4210 4215 4220
Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Glu Arg
4225 4230 4235 4240
Val Ile Arg Gln Ala Trp Ala Asn Ala Gly Val Ala Ala Met Asp Ile
4245 4250 4255
Asp Ala Val Glu Gly His Gly Thr Gly Thr Thr Leu Gly Asp Pro Ile
4260 4265 4270
Glu Ala Gln Ala Leu Leu Gly Thr Tyr Gly Gln Gly Arg Ser Ala Asp
4275 4280 4285
Arg Pro Leu Trp Leu Gly Ser Ile Lys Ser Asn Val Gly His Thr Gln
4290 4295 4300
Ala Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Met Ala Met Arg
4305 4310 4315 4320
His Gly Leu Leu Pro Gln Thr Leu His Ala Glu Glu Pro Ser Pro His
4325 4330 4335
Val Asp Trp Ser Gly Gly Thr Val Arg Leu Leu Thr Glu Pro Val Ala
4340 4345 4350
Trp Pro Glu Arg Gly Arg Met Arg Arg Ala Gly Val Ser Ser Phe Gly
4355 4360 4365
Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Gln Ala Pro Pro Asn
4370 4375 4380
Ala Glu Thr His Glu Pro Ala Glu Pro His Thr Ala Pro Gly Pro Leu
4385 4390 4395 4400
Pro Trp Thr Ile Ser Ala Lys Ser Pro Gln Ala Leu Arg Ala Gln Ala
4405 4410 4415
Arg Gln Leu His Thr Tyr Leu Thr Asn Thr Pro Glu Ala Asn Pro Ala
4420 4425 4430
Asp Val Gly His Thr Leu Ala Met Gly Arg Ala Ser Phe Glu His Arg
4435 4440 4445
Ala Val Val Ile Gly Ser Asp Arg Gly Glu Phe Leu Gly Gly Leu Asp
4450 4455 4460
Ala Val Ala Ala Asp Glu Ala His Ser Ala Val Val Thr Gly Ile Ala
4465 4470 4475 4480
Arg Lys Ala Gly Asp Leu Gly Lys Val Val Phe Val Phe Pro Gly Gln
4485 4490 4495
Gly Gly Gln Trp Ala Gly Met Gly Leu Arg Leu Leu Lys Thr Ser Pro
4500 4505 4510
Val Phe Ala Gln Ser Ile Gln Ala Cys Glu Gln Ala Leu Ala Pro His
4515 4520 4525
Thr Asp Trp Thr Leu Thr Asp Ile Leu His Arg Pro His Thr Asp Pro
4530 4535 4540
Leu Trp Gln Arg Ala Asp Val Ile Gln Pro Ala Leu Phe Ala Leu Met
4545 4550 4555 4560
Thr Ser Leu Thr Thr Leu Trp Gln Ser His Gly Leu Asn Pro Asp Ala
4565 4570 4575
Val Ile Gly His Ser Gln Gly Glu Ile Thr Ala Ala His Ala Cys Gly
4580 4585 4590
Ala Leu Ser Leu Glu Asp Ala Ala Lys Ile Val Ala Leu Arg Ser Gln
4595 4600 4605
Thr Leu Gln Thr Leu Gln Gly Ser Gly Gly Met Ala Ser Val Pro Leu
4610 4615 4620
Pro Ala Asp Gln Val Thr Ala Leu Leu His Thr Met Trp Pro Asp Gln
4625 4630 4635 4640
Leu Trp Val Ala Ala Ile Asn Ala Pro Thr Thr Thr Val Ile Ser Gly
4645 4650 4655
Asp Thr Gln Ala Leu Thr Gln Ala Leu Asn His Tyr Arg Asp Gln Asp
4660 4665 4670
Ile Asp Ala Lys Arg Ile Pro Val Asp Tyr Ala Ser His Cys Pro His
4675 4680 4685
Ile Gln Ala Val Gln His Glu Leu Ser Asp Leu Leu Gln Asp Ile Thr
4690 4695 4700
Pro Arg Ala Ala Thr Thr Pro Phe Tyr Ser Thr Thr Asp Asn Gln Trp
4705 4710 4715 4720
Thr Asp Thr Thr Thr Leu Asn Ala His Tyr Trp Tyr Arg Asn Leu Arg
4725 4730 4735
Gln Pro Val His Leu Thr Asn Ala Ile Thr Asn Leu Thr His Gln Gly
4740 4745 4750
His His Thr Tyr Ile Glu Ile Ser Pro His Pro Thr Leu Thr Pro Ala
4755 4760 4765
Ile Gln Glu Thr Thr His Thr Thr His Thr Pro Thr Thr Val Ile Ser
4770 4775 4780
Thr Leu Arg Arg Asn His Asn Asp Thr His Gln Leu Leu His Ala Leu
4785 4790 4795 4800
Ala His Ala His Thr Thr Gly His Pro Ile Asn Trp His Pro Thr His
4805 4810 4815
Gln His His Thr Pro Thr Pro Gln His Thr Asp Leu Pro Thr Tyr Pro
4820 4825 4830
Phe Gln His Gln Arg Tyr Trp Leu Asn Thr Pro Thr Gln Thr Gly Asp
4835 4840 4845
Ala Ala Ala Ile Gly Leu Asp Pro Ala His His Pro Leu Leu Gly Ala
4850 4855 4860
Ala Val Ala Val Ala Glu Gly Glu Gly Tyr Leu Leu Thr Gly Arg Leu
4865 4870 4875 4880
Ala Leu Ser Thr His Pro Trp Leu Ala Asp His Thr Ile Ala Gly Ala
4885 4890 4895
Val Val Leu Pro Gly Thr Ala Leu Leu Glu Ile Ala Leu Gln Ala Gly
4900 4905 4910
His Arg Val Asp Cys His Arg Ile Glu Glu Leu Thr Leu Gln Ser Pro
4915 4920 4925
Leu Phe Ile Pro Glu Glu Gly Ala Val Gln Val Gln Ala Trp Val Ala
4930 4935 4940
Ala Pro Asp Glu Asn Gly Tyr Arg Ser Leu Thr Val Ser Ser Arg Arg
4945 4950 4955 4960
Glu Gly Thr Tyr Glu Asp Ala Thr Trp Val Arg His Ala Thr Gly Arg
4965 4970 4975
Val Gly Pro Ala Pro Ala Asp Gln Asp Asp Ala Ile Ala Arg Leu Thr
4980 4985 4990
Asp Pro Gln Gly Asp Gly Ala Ala Ala Val Trp Pro Pro Gln Gly Ala
4995 5000 5005
Val Ala Phe Thr Ala Asp Asp Leu Glu Gly Leu Tyr Asp Gly Tyr Ala
5010 5015 5020
Ala Arg Gly Phe Glu Tyr Gly Pro Val Phe Arg Gly Leu Arg Ala Ala
5025 5030 5035 5040
Trp Arg Arg Gly Glu Asp Ile Phe Ala Glu Val Arg Leu Pro Asp Thr
5045 5050 5055
Ala Asp Gly Asp Ala Ser Gln Phe Ser Val His Pro Ala Leu Leu Asp
5060 5065 5070
Ala Ala Leu His Ala Ala Ala Phe Arg Pro Ala Asp Glu Leu Pro His
5075 5080 5085
Gly Ala Leu Pro Phe Ser Phe Ser Gly Val Arg Leu His Gly Pro Gly
5090 5095 5100
Ala Ser Thr Leu Arg Val Arg Leu Thr Pro Asp Gly Gln Ala Arg Asp
5105 5110 5115 5120
Thr His Ala Trp Ser Val Ala Val Val Asp Gly Glu Gly Arg Pro Val
5125 5130 5135
Ala Ser Ile Ala Ser Leu Ala Val Arg Pro Val Ser Thr Gln Glu Leu
5140 5145 5150
Leu Ala Ala Ser Gly Thr Ala Arg Arg Asp Ser Leu Phe Ala Val Glu
5155 5160 5165
Trp Val Thr Ala Pro Ala Pro Thr Ser Ser Ser Ala Pro Arg Arg Leu
5170 5175 5180
Ala Thr Val Gly Pro Ser Asp Arg Leu Pro Ser Ala Asp Ala Tyr Ala
5185 5190 5195 5200
Asn Leu Ala Asp Leu Ala Ala Ala Val Leu Glu Ala Glu Ala Pro Ala
5205 5210 5215
Pro Asp Ala Val Val Val Asp Cys Gly Arg Arg Asp Ala Arg Ala Thr
5220 5225 5230
Ala Val Ala Glu Asp Val Arg Thr Leu Thr Arg Arg Ile Leu Gly Leu
5235 5240 5245
Leu Gln Glu Trp Leu Ala Asp Glu Arg Pro Ala Ser Ser Arg Met Val
5250 5255 5260
Val Leu Thr Arg Gly Ala Val Ala Thr Thr Pro Gly Glu Asp Val Ala
5265 5270 5275 5280
Asp Leu Ala Gly Ala Ala Val Cys Gly Met Val Arg Ser Ala Gln Ser
5285 5290 5295
Glu His Pro Gly Arg Phe Val Leu Leu Asp Leu Asp Pro Asp Pro Asp
5300 5305 5310
Leu Asp Gly Gly Glu Val Pro Pro Thr Val Val Pro Ala Ala Leu Ala
5315 5320 5325
Cys Gly Glu Pro Gln Ile Ala Val Arg Ala Asn Arg His Leu Val Pro
5330 5335 5340
Arg Leu Thr Arg Val Pro Val Ser Val Pro Val Pro Gly Pro Val Pro
5345 5350 5355 5360
Val Pro Ala Ala Glu Ala Ala Asp Gln Asp Thr Thr Pro Thr Ala Phe
5365 5370 5375
Asp Pro Asp Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Thr Leu Gly
5380 5385 5390
Ala Val Leu Ala Arg His Leu Val Ser Arg His Gly Val Arg His Leu
5395 5400 5405
Leu Leu Ala Ser Arg Arg Gly Pro Asp Ala Pro Gly Ala Thr Glu Leu
5410 5415 5420
Arg Ala Glu Leu Ala Glu Leu Gly Ala Glu Ala Thr Val Arg Ala Cys
5425 5430 5435 5440
Asp Thr Gly Asp Arg Gly Ala Leu Ala Asp Leu Ile Ala Gly Ile Pro
5445 5450 5455
Thr Gly His Pro Leu Thr Gly Val Val His Ala Ala Gly Val Leu Asp
5460 5465 5470
Asp Ala Thr Val Ala Ser Leu Thr Pro Arg His Leu Asp Thr Ala Leu
5475 5480 5485
Thr Pro Lys Ala Asp Ala Ala Phe His Leu His Glu Leu Thr Arg His
5490 5495 5500
Ala Arg Pro Arg Ala Phe Val Leu Phe Ser Ser Ala Ala Gly Val Leu
5505 5510 5515 5520
Gly Ala Ala Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Phe Leu Asp
5525 5530 5535
Ala Leu Ala Glu His Arg Arg Ala Gln Gly Leu Pro Ala Leu Ser Leu
5540 5545 5550
Ala Trp Gly Leu Trp Glu Gln Gly Ser Gly Met Thr Gly His Leu Asp
5555 5560 5565
Arg Thr Asp Arg Ala Arg Ile Asn Arg Ser Gly Leu Ala Pro Leu Ala
5570 5575 5580
Thr Glu Asp Ala Leu Ala Leu Phe Asp Ala Ala Leu Ala Gly Asp Arg
5585 5590 5595 5600
Pro Phe Leu Val Pro Ala Arg Leu Asp Leu Arg Gly Ser Ser Ala Ala
5605 5610 5615
Glu Thr Pro Ala Pro Leu Phe Ser Arg Ile Ala Pro Ala Arg Thr Thr
5620 5625 5630
Arg Gly Arg Thr Pro Gly Ala Glu Gly Ala Ala Asp Leu Arg Thr Arg
5635 5640 5645
Leu Ala Ala Gln Asp Ala Thr Glu Gln Arg Asp Thr Leu Leu Thr Ile
5650 5655 5660
Val Arg Thr His Thr Ala Ala Val Leu Gly His Asp Thr Ala Ala Ala
5665 5670 5675 5680
Val Arg Pro Asp Ala Ala Phe Arg Glu Leu Gly Phe Asp Ser Leu Ala
5685 5690 5695
Ala Val Glu Leu Arg Asn Arg Leu Gln Thr Thr Thr Ala Leu Thr Leu
5700 5705 5710
Pro Ala Thr Thr Val Phe Asp His Pro Thr Pro Ala Ala Leu Ala Asp
5715 5720 5725
His Leu Arg Thr Gln Leu Cys Gln Asp Ala Pro Ser Pro Ala Ala Ala
5730 5735 5740
Thr Ala Met Ala Ala Met Ala Glu Leu Ala Arg Leu Glu Ser Ala Val
5745 5750 5755 5760
Ser Asp Ser Ala Ala Leu Asp Asp Asp Thr Arg Ser Gly Leu Ala Glu
5765 5770 5775
Arg Leu Arg Ser Leu Ala Arg Lys Met Ser Ser Gly Arg Val Val Asp
5780 5785 5790
His Asn Gly Gly Gly Ala Ala Gly Leu Asp Leu Gln Ser Ala Thr Asp
5795 5800 5805
Asp Glu Met Phe Glu Leu Ile Asp Lys Glu Val Ser Arg Asp
5810 5815 5820
<210> 77
<211> 17481
<212> DNA
<213> Artificial Sequence
<220>
<223> milA3 gene of Streptomyces bingchenggensis
<400> 77
atggccgctg gccacgacaa ggtgatcgag gcgctgcggg cgtccctcaa gaccaacgag 60
cggcagaggg aacagatcca ccggctcact acggcggcgc gggaacccat cgccatcatc 120
ggcatggcct gccgctatcc gggcggagtg ggatcgccgg aggacctgtg ggagctggtg 180
gccgccggtc gtgacgccat cggcaccttc cccgaggacc ggggctggga cgtggagcgg 240
ctgtacgacc ccgatccgga gcgggccggc acctcgtgta cccagcatgg cggattcctg 300
taccaggcag gggagttcga ccccggtttc ttcgggatca gcccgcgcga ggcgctggcg 360
atggacccgc agcagcggct gctgctggag atctcctggg aggtgttcga gcgggccggg 420
atcgacccgg cctcggtgcg cggcagccgc accggggtgt tcgcgggcgt catgtaccac 480
gactacggct cccggctgca caccgtcccc gaaggcttcg agggctatgt cggcaacggc 540
agcggcggcg gcgtggcgtc cggccgggtc gcctacaccc tcggcctcga aggcccggcc 600
gtgaccgtgg acaccgcctg ctcctcctcg ttggtcgccc tgcacctggc ctgccaggcg 660
ctgcgggccg gcgagtgctc actcgccctg gcgggcgggg tgacggtgat gtccaccccc 720
agcctgttcg tcgagtactc ccggcagcgc gcgctcgcgg cagacggccg gtgcaaggcg 780
tacggggcgg gggcggacgg caccggctgg gcagaaggcg ccgggatgct gctggtggaa 840
cggctcacgg acgcacagcg cctcggccac cgggtgctgg cggtggtccg gggcagcgcg 900
gtcaaccagg acggcgcgag caacggcctc accgccccca acggccccgc gcaacaacgg 960
gccatccggc aggcactggc gagcgccggg gtgtcggcgt ccgaggtcga cgccgtggag 1020
gggcatggga cggggacgcg gctgggcgat ccgatcgagg cgcaggcgtt gctggcgacc 1080
tacggtcagc agcggcccgc ggaccggccg ctgtggctcg ggtcgatgaa gtccaacgtc 1140
ggccatgcgc aggcggccgc cggcgtgggc gggatcatca agatggtgat ggccatgcgg 1200
agcgggacgc tgccgcgcac cctgcacgcg gacgagccgt cgccacacat cgactgggac 1260
tcgggcgcgg tacggctgct gaccgagccg gtcgcctggc cggagcgcga ccggccccgc 1320
cgcgccgcgg tgtcctcctt cggggtcagc ggcaccaacg cccatgtgat cctcgaggcc 1380
gcatcgcaga cggcgccgca gacggattcc gcgtcgcagg cggaaaccga cgacgctccc 1440
gcaccgcacg gcgcgccggg ccatgccgtg gcggggccgc tgctctggcc cttgtcgggc 1500
gcgacggccg aggcgctgcg ggcccaggcc ggggagctgc gtcgcttcgt ggcggccgat 1560
gagctgctgc gccccgccga cgtcgggcac accctggtct tcggccgctc ggacctcgca 1620
caccgcgcag tcgtcctcgg ctccgaccgg gaaaccctgc tgcgcgctct ggacactctg 1680
gcaggggagg ggccggacga cggctcggtc gtacggggca tggcggccgc cggggccggt 1740
gcgggcgtgg tgttcgtctt cccgggacag ggcggccagt gggccggcat ggggctgcgg 1800
ctgctggaga cctcgtcgtt cttcgccgag cggatggcgg agtgcgaggc ggcgttggca 1860
ccgtatgccg actggtcgct gctcgacgtt ctgcgccggg accccgggga cccggtctgg 1920
gagcgggccg atgtcgtcca gccgatgctg ttctcggtga tggtgtcgct ggcgcagctg 1980
tggcgctcgt acggcgtcga accggacgcc gtactcggcc actcccaggg cgagatcgcc 2040
gccgcccaca tctgcggcgc gctgaccctg gacgacgccg cgaaggttgt cgcgctgcgc 2100
agccgggccc tgcagaccct gcgcggttcg ggcggcatgg cctccgtacc actgccggcg 2160
gacgaggtca ccgggctgct gcggaccgac tctctgtggg tggccgccgt caacgccccc 2220
acggccacgg tgatctccgg cgacgcggac tctctggcgg aggcgctgga acactaccgg 2280
gaccagggcg tcgaagcgaa gcgggtcccg gtcgactacg cctcccactg cccgcatatc 2340
gaagccgtgg agcaggagct gctgggcctg ttgcggggga tcgctccaag ggccgccgac 2400
atccccttct actccaccgt ggacaaccag tgggccgaca ccatgggact cgacgcccgg 2460
tactggtacc gcaatctgcg ccggcccgta cgcttcgccg aagcgctccg cgccctcggc 2520
gccgccgagt accggacgta tgtcgaggtc ggcccgcacc ccaccctcac ccccgccatc 2580
gaggacacca ctgaggccgc cggcgtcgcg gccacggttg tcggatccct gcgccgcggc 2640
gaggacgacg cccaccgcat cctgacctcg ctggcccggg ctcatattca tggcctgccc 2700
gtggcgtggg accgccacta ccgggcgctc gcccccgagg cgaaccatgt cgacctgccc 2760
acctacgcct tccagcgccg ccgctactgg ctggacgccc cggcgaccac cggggacgtg 2820
acggccgcgg ggctggcccc ggtcggacac ccactgctcg gcgcggcggt cggactcgcc 2880
gagggcgacg gatatctgct caccggccgg ctcgccccgc acacccaccc ctggctcacc 2940
gaccacgcgg tcgccggcac cgtcctgctg ccgggcaccg catacgtgga actggccgtg 3000
cacgtcggcg gacacctcgg ctgcccccgg ctggaggagc tcaccctgca cgccccgctc 3060
gtcctccccg acaccggcgg cgtggcgctc caggtggccg tcggggcacc ggacgagacc 3120
ggccgccgcg cactgagcgt ctacgcacag cgcgacgacg accccgcgtg ggagggggcg 3180
gcccggggcg cgtggacacg gcatgcgacc ggcacactgg cggccgaggc cccgactgat 3240
ggcatcagcg gtgccgacgg tgccgggacc ctggcggggg cgtggcctcc gccgggcgcg 3300
gagcccctgg acatcagcgg cctctacgac acgctggccg ccgcagactt cggctacggc 3360
ccggccttcc aggggctgcg cgccgtctgg cggcaaggcg aggagaccta cgccgaggtg 3420
cggctccccg accaggtggc cgccgacgcc ccacgcttct gcctccaccc cgcgctgctc 3480
gacgccgcgc tccacccgct ggcactcgac agcggccgaa gcgaggagaa tccagcggga 3540
catggcctgc tgccgttcgc ctggcgcggc gtcagcctgc gctccccggg cacaccgacg 3600
ctgcgcgtac ggctgcggcc gcagggcccg gactcgattg ccgtcgacgt ggccgacgag 3660
acgggcgcgc cggtggcctc ggccgaatcg ctcacgctgc ggccggtggc cctggaggac 3720
ctgcgggccc tcggcggcca ggcgggcgac accctctacg ccctggagtg gaccgccgcg 3780
cccgagcccc cggcgacggc cctcgggcgg tgcgctgtga ttggccaagc cattcctgga 3840
tgggctgccg cgctggagac ggcggcagcg gggcccgtac ggcggtaccc ggaccttgcc 3900
ggactggtga cggccctgga cgcgggcgat ccgcctccgg acctggtgtt cgtgggctgc 3960
cctccggctg ccgccgggcc cgacgacacg acggtcgccg acgtccacac cgcccgtacc 4020
cgtgtccgta cccgacaagc gctggacctg cttcagggct ggctcggcga agcgcggctg 4080
gccggcgcga ggctggtgct ggtcacctgc ggcgcggtgg ccaccgggcc ggcggaggga 4140
gtgatggacc tggcgggcgc ggcgatctgc ggactggtgc gatccgcgca ggccgaggag 4200
cccgaccgta tcctcctggt ggacctggac gcggccgagg agtcgtgggc ggcgctacca 4260
cgggcggtcg cgctgggcga accgcagatg gccatccggg ccggccagcc gcacatggcc 4320
cggctggttc gagccgacac cgaggggggc gccctgctca cgccgccaca ggggagcggc 4380
ggctggcggc tcgactgcgc cgacgcgggc acggtccagg ggctggcgcc tgtggcgtcc 4440
tcggccgacc gcgacccgct gggcccgcac caggtacgga tcgaggtgcg tgcggccggg 4500
ctgaacttcc gcgatgtcct ggtggccctg gggatggtcc ctgggcagcg ggggctgggc 4560
agcgagggcg ccggggtggt gctcgaagcc gggcctgaag tggccgacct ggcgcccggg 4620
gaccgggtga tgggcgtgtt cgcggatgcg ttcggcccgt tcgcgatcgc cgaccgggcc 4680
accgtgatcc gcgtccccga ccactggacc ttcggccagg ccgccgccgt ccccgtcgtg 4740
ttcgccaccg cctattacgg gctggtggac ctggcaggac tgcgcccggg tgagtcggtg 4800
ctggtgcacg ctgcggccgg cggagtggga ctggccgctg tccaactggc ccgccacctg 4860
ggcgctgagg tctacgccac ggcgagcccc ggcaaatggg acaccctacg cgcccacggc 4920
atccccccgg agcgcatcgc ctcgtcccgc accctcgact tcgagagccg gttcaccggc 4980
cggaacatcg acgtcgtcct caactccctg gcccatgagt acgtcgacgc ctcgctgcgc 5040
ctggtgtccg gcgacagcgg ccggttcctg gagatgggca agaccgacct ccgcgacccg 5100
gaggaggtgg cgcaggcgta ccccggtgtc gcctaccggg cgtacgacct gatggaggcc 5160
ggacccgagc gcatcgggga gatcctgcgc accgtgttgc ggctgttcga cgagggcgtg 5220
ctcaccccgc tgccgctcac ctgctgggac atccggcagg ccagggatgc cttccgccaa 5280
ctccagcagg gccgcaccgt cggaaagaat gtgctcacgc tggaccgcac ccccgacccc 5340
gacggcaccg tcctcatcac cggtggcacc ggtaccctcg gcgccgcgct cgcccgccat 5400
ctcgccgcca ccggccgagc acggcatctg ctactgatca gccgccgtgg cctcgatgcg 5460
ccaggcgctc ccgaactcat cgctgagatt gacgagttgg gcgccacggc gaccgtcgcc 5520
acctgcgacg tcggcgaccg tgccgcgctc gccgaactgc tcgggcggat ccccgccgag 5580
cacccgctga ccgccgtcgt ccacgccgcg ggcaccctcg acgacgccac gctcggctcc 5640
ctcaccgcgc gccacctcga caccgttctg cccgcgaagg ccgatgccgc ctggcatctg 5700
cacgacctga cctgccggct ggatctggcc gcgttcgtgc tgttctcgtc cgccgcgggt 5760
gtcctgggct cgccggggca gggcaactac gccgccgcca acgcctttct cgacgcgctc 5820
gccttccagc gacgggcgat gggactcccc gccgtgtccc tggcatgggg actgtgggag 5880
gaggccagcg gaatgaccgg ccacctcgac cagaccgacc gcacccgcat ggcccgcgtc 5940
ggcctccggc cactggccac ggacgaggcc ctggcgctgt tcgacaacgc tctcgtcgac 6000
ggcccaccgc tgctgctccc ggcccgtatc gacaccaagg cgctacgggg caccaccgca 6060
ccgcccctgt tccagagcct cgtacgcccc accaccggcc accggccacg ccccgcgaca 6120
cccgacggcc gctcctccct ccgagcccgg ctcgccgggc tcgaccccgc cgcacagcac 6180
gaggtcctgc tcaccctcgt ccgcggccac gccgccacgg tcctcggcca cccgagcccc 6240
gacgccatcg cccgcgaggc ggccttccgt gacctcggct tcgactccct caccgccgtg 6300
gagctccgca accgcctcaa ggaggcaacc ggcctgcggc tccccccccc cccccgcctc 6360
aaggaggcaa ccggcctgcg gctccccgcc accatcgtct tcgaccatcc cactcctgcc 6420
gctctcgccc agcacctgcg ggacggcctc atcggcggcg ccgatacggt caccctggct 6480
gcggctcctg ctccgagcaa ggtggcgatg gtggcggatg aggccatcgc gatcatcggc 6540
atggcctgcc ggtatccggg gggcgtgcgg tcggccgagg ggctgtggga tctggtcgcc 6600
tccggcaccg acgccatgag cggattcccc agcgaccgcg gctgggacct cgaccgcctc 6660
tacgcccccc aggaccagga cgtgccgggc accacataca cccgccacgg gggcttcctc 6720
cacgacgcgg gcaagttcga cgcgggattc ttcggcatcg gcccacgtga ggcgctggcg 6780
atggatccgc agcagcggct gctgctggag acctcctggg aggttttcga acacgcggga 6840
atcgacccct cgtcggtacg gcggagccgg accggagtct tcgccggtgt gatgccgacg 6900
gactacggcc cccggctgca agacaccgtg gccgaggtcg agggctatgt cctcaccgga 6960
aactccggca gcgtcgcctc gggccgtatc gcctacacct tcggcctgga aggccccgcg 7020
gtgtcggtgg acacggcgtg ttcgtcgtct ctggtggcgt tgcatctggc gtgtcaggcg 7080
ctgcgtgcgg gggagtgctc catggcgctg gccggcgggg tgacggtgat ggcgacgcct 7140
ggtgccttcg tggagtttgc gcggcagcgg gggttgtcgg tggatgggcg gtgcaaggcg 7200
tttggggtgg gtgcggatgg tacggggtgg gcggaggggg tggggatgct gttggtggag 7260
cggttgtctg atgcgcggcg gttggggcat cgggtgttgg cggtggtgcg gggttctgcg 7320
gtgaatcagg atggtgcgtc gaatggtttg acggcgccga atggtccgtc gcagcagcgg 7380
gtgatccggc aggcgttggc cagtgcgcgg gttggcgggg cggatgtgga tgtggtggag 7440
gggcacggta cggggacgcg gctgggtgat ccgatcgagg cgcaggcgtt gctggcgacc 7500
tacggtcagg aacgccctga tgatcgacct gtctggttgg ggtcggtgaa gtcgaatatc 7560
gggcatgcgc aggccgcggc gggggttgcg ggtgtcatca agatggtgat ggcgatgcgg 7620
tatggggtgt tgccgcggac gttgcatgtg caggagccgt cgccgcatgt ggactggtcc 7680
tcgggcgggg tgcggctgct gacggaggcg gtgccgtggc cggagacggg gcgtgcgcgg 7740
cgtgcggggg tgtcgtcgtt cggggtcagt ggcaccaacg cgcacatcat cctcgaacag 7800
gcgccgcctg aggagcacga cgatccggcg gacgtctcgt ccgggtcgtt tccgtggatg 7860
gtgtcggcca agtccgaaca ggcactacag gcgcaggcag cacagttgcg cgcgtatctg 7920
gcggcacatc ctgagctggg gctggctgat gtcgggtatg cgctggcctc cggccgcacg 7980
gccttcggcc accgtgccgt gctcctgggc ccggaccgcg aagccttcgt cgaagagctg 8040
ggagctctgg aggccggtga ggaacacgcc gggctggtac ggggcgtggc gacgggtgcg 8100
gggaagctgg cgtttgtgtg ttccgggcag ggaacgcaac gtccccgtat gggacacggg 8160
ctgtactcgc cttcccgctg ttcgccgcag ccatggacga agcctgcgca cacctggacc 8220
cacacctcga ccatcccctg cgggatgtca tgttcgccga gccgggcacc gacaccgccc 8280
agctgctcca ccagacccgc tacgcccagc ccgcgctgtt cgccctccag gtcgccctgc 8340
accgcctggt caccgaacac cacggcctta ccccccacta ctacgccggc cattccctcg 8400
gagagatcac cgcggcccac ctcgccggga tcctcaccct ccccgacgcg gcccgcctgg 8460
tcaccacccg cgcccgcctc atgcaatctc tccccgccac cggcgccaat gaccaccctc 8520
caagcagacc ccgacgaact ccacgaacac ctcacacgat gcgaaggacg ggtctcactc 8580
gcggccgtga acgcgcccgg gtccgtggtc atcagcggtg atcgccacga cgtagacgct 8640
acggccgaaa acctccgcgc catgggacgc aagaccactg cgctgaaggt cagcggcgct 8700
ttccactcac accacatcga cccactcctc aacgaactcc gcaacacggc agaaaccctc 8760
acctaccacc caccccacac ccccctcatc accaccaacc ccaccgacca cgaccccacc 8820
acaccccact actgggtccg gcaagcgcgc gagacggtcc actacgccca caccacccaa 8880
caactccaca cccacggcgt caccgcctac ctcgaactcg gccccgacca caccctcacc 8940
gccctcaccc accacaacct ccccgaccac accccgctag ccgtcccgct tctccacccc 9000
gaccaatccg agacccacac cacccacacc gccctcgccc acctccacac ccacggccac 9060
cccaccacct ggcaccacca tcacaccccc acccactacc acccaaacct ccccacctac 9120
cccttccaac accaccacta ctggctcaac accaccactg ccaccggtga tatgtcggct 9180
gcaggccttg agccggcgcg gcatcccctg ttgggcgcgg cggtcgggtt ggccgatggt 9240
gaggggttgc tgttcactgg gcggatttct ctccgtacgc atccctggct ggccgaccac 9300
gccgtcggcg gcgccgtgtt gctccccggt acggcctttc tcgaactcgc cctccaagcc 9360
gccgcccatg ccgactgccg tcgggtcgag gagcttacgc tccacacccc gctcgtcgta 9420
ccggatagcg ccggcgtagt gctgcaggtc actgtggccg cgccgaacga agcaggaaac 9480
cgggcggtgg atatctactc gcgaatcgat gtcggcggcc tcaccgccga ttcggctggc 9540
gagccgtgga cgcgccatgc cgccgggtac cttgccgaca agcctgaccc agactgcggt 9600
gactcggcgg atggtgtcat gcccgcgggc gcatggccgc cgccgggtgc ggtcgccgtg 9660
gatctggagg gactgtacga gcaactggcc gaggggggtt tccactacgg tgcggccttc 9720
cgttgcctgg acgccgcctg gcaacgcggg gacgaggtct tcgcgaccgc gtatatgtca 9780
gaggatcagc tgggcgacac ggctgcggct cggttcgcgc tgcaccccgc gctgctggat 9840
tccgcactgc acaccattcc acttttgccc tccctacggg gacaacagga cagcgggctg 9900
ccgttcacgt ggacaggagt caccctgcgt gcatccgggg cgacggctct gcgcgtccgg 9960
ctgaggccgg acggccatgg cccgggggcg gtgtcggtcg acgtgtccga cgaggcgggt 10020
gagcccgtag catcggtccg gtcgttggcc ctgcggccgg tgaccagggc cgagttgcat 10080
acggccgagt tgcgcacagc cgccccggtt gccccccatg gctcgctctt cgaggtgcga 10140
tgggaacccg tcccccagcc ttcagcggcc gaagaagccg ccccatgggt gatgatcggg 10200
accgggccga cgctgcgccc ggtcgaggac ttcgtcactc cgccggagcg gacgtacgcc 10260
gacctggccg cgctgtgcgt ggcaatcgcc gatgacgcgc ccgttccccg gacggtcgtg 10320
gcctggtccc cagccgggag cgaagacgag tcgagtgagg cgctgcgcca ggccacacac 10380
cacatgctgg gcctactgca gcagtggttg gcggacagcc ggttcgccga cagtcgcctg 10440
gtgatcctca cccgagccgc ggtggccact gcgccggacg aggaggtaga agacctggcg 10500
ggagcggcgg cgcggggtct gatccgctcc gcccagtcgg agcaccctga ccgattcgtc 10560
ctgctcgacc tggacgaccg tcccgctgac gcgaaagacc acgaccgaat gctgtcgatg 10620
gccctggcct gcggggaacc ggaagtggcc gtacgcgatg gagccctgcg cacaccccgg 10680
ctgagcccgc tggccggcac cgccaccgag gccatggacg agcatccctg ggatcaggac 10740
ggcaccgtac tcatcaccgg cggcaccggc agcctcggcg ccatgcttgc ccgccacttg 10800
gtggccaccc atggcgtacg gcatctgatg ctgatcagcc gacgtggcct cgacgccccg 10860
ggggccaggc gactgggggt cgaacttgcg gagctcgggg cgcaggtgac gatcaccgcg 10920
tgcgatgccg cagaccaaag gcaacttgcg aacgtattgt cggagatctc cgtcgaccat 10980
ccgctgaccg ctgtggtgca tgcggcaggc gtactggacg acggggtgat cacatccctc 11040
acaccggagg gcctgaccca tgtcctgcgg gccaaggtcg attcggcgct caatctccac 11100
cagctcacac gcgacctgcc gctgtccgcg tttgtgctct tctcctcgct ggccggggtg 11160
atgggttcgg cagggcaggg caactacgcc gccgccaacg cagccctgga cgcgctggcg 11220
agtcaccgga gggccgctcg gctgccggcg gtgtccttgg cctggggagt ttgggagcag 11280
accgagggca tgaccgggca gttggaggcc acggaccacg cgcggctccg ccgctcgggc 11340
ctgaggccgc tggccatcag cgagggcctg gagctcttcg acaaggccct gagctgtgga 11400
cacgccctgg tggtgcccgc cgcactcagc acgagggagc ttcagacatc cggatccgtc 11460
ccgccattcc tgcgccacct gacgggtgtc gctccggccc ggccgtcccg gacccgcgac 11520
gcctcggccg gtgagccgac ctccctgcgg cggcggttga ccggcctcgg gccggaagaa 11580
cggctacgcg aggtgctgcg gctggtgcgc tcccgggcgg ctgcggtgct ggggcacggc 11640
acggccgaat cggtcccggc ggactcggcg ttccgcgacc tggggttcga ctccctcgcc 11700
gcggtggacc tgcggaaccg gttgcagcag gccaccgggc tgcgcctgcc ggccggcttg 11760
atcttcgacc ggccgcgtcc ggacgtgctc gcccgtttcc tgtgtgacga gttggccggc 11820
gccggcggta cgtcggcggc cacggccgcc ccacccgttg cggccggcgg gggggggggc 11880
cgcgggggag ccggtggcca tcgtcggcat ggcatgccgg tttccgggag gtgtgcggtc 11940
ggccgagggc ctgtgggatc tggtcgcctc cggtatggac gcgtgggtga cttccccgca 12000
gaccgaggct gggaggtgga acggctctac gaccccgacc cggaccgaac cggcacctcc 12060
tacacccggc aaggcgggtt cctctacgac gcgggtgagt tcgacgcggc attcttcggg 12120
atcggcccgc gtgaggcggt agccatggat ccacagcagc ggctgctgct ggagatctcc 12180
tgggaggcgc tggaacgtgc ggggatcgac ccggcgtcgc tgcgggggag ttcgaccggg 12240
gtgttcgctg gggtgatgta ccacgactac ggcacccgcc tgcgcgagat cccagagggc 12300
tacgagggct atatcggcaa tggaaacgcg ggcagcgtcg cgtcgggacg tgtcgcctac 12360
accttcggcc tggaggggcc ggcggtcacc gtggacacgg cgtgttcgtc gtccctggtc 12420
gccctgcatc tggcctgcca ggcgctgcgg tcaggggagt gctccatggc gctggccggc 12480
ggggtcaccg tcatgtccac ccccaccact tttgtcgagt tctcgcgcca gcggggactg 12540
gccccggacg ggcggtgcaa gtccttcggg gccggcgcgg acggaacagg ctgggcggag 12600
ggggcgggga tgctcctggt ggaacggctt tcggacgccc ggcgcaacgg ccaccgggtc 12660
ctggcggtgg tacgggggag tgcggtcaac caggacgggg cgagcaatgg gctgacggcg 12720
ccgaacggcc cgtcgcaaga gcgggtgatc cgccaggcgt gggcaaacgc gggtgtggcc 12780
gcgatggaca tcgacgcggt ggagggacac ggcacgggga cgacgctcgg tgaccccatc 12840
gaggcccagg cgctgctggg gacgtacgga cagggacggt cggccgatcg gccgttgtgg 12900
ttgggatcga tcaagtccaa cgtcggacac acccaggccg ccgcgggggt gggcggcgtc 12960
atcaagatgg tgatggccat gcgccacggg ctgctcccgc agaccctgca cgccgaggag 13020
ccctcacctc atgtggactg gtcgggcggg acggtgcggt tgctgaccga gtcggtggcc 13080
tggcccgagc aggggcggat gcgccgtgcg ggcgtctcct ctttcggtgt cagcggtacc 13140
aacgcccacg tcatcctgga acaagcaccg cctgccgcgg agacccacga accggcagag 13200
cccaacaccg cgccaggccc actgccctgg gcgatctccg cgaagagccc gcaagcgcta 13260
cgtgcccagg cgcgccaact gcacacgtac ctgaccaacg cccccgaggc gaaccccgcc 13320
gacgtcggcc acaccctcgc gacgggccgc gcctctttcg agcatcgtgc tgtggtcatc 13380
ggctccgacc gagcggagtt cctgggtggc ctggatgctc tggcggccga cgaggcccac 13440
accgccgtcg tcacggggat cgcgaggaag gccggtgacc agggcaaggt ggtgttcgtg 13500
ttccccgggc agggcggtca gtgggccggg atgggactgc ggctgcttaa gacctcaccc 13560
gtcttcgccc aatcgatcca ggcctgcgaa caagccctcg ccccccacac cgactggacc 13620
ctgaccgaca tcctgcaccg gccccacacc gaccccctgt ggcagcgcgc cgacgtcatc 13680
cagcccgtcc tcttcgccct catgacctcc ctcgccgccc tctggcaatc ccacggcctt 13740
aaccccgacg ccgtcatcgg ccactcccaa ggcgaaatca ccgccgccca catcagcgga 13800
gcgctgagcc tggaggacgc cgcgaaaacc gtcgcgctgc gcagccgggc cctgcagacc 13860
ctgcgcggtt cgggcggcat ggcctccgta ccactgccgg cggacgaggt caccgggctg 13920
ctgcggaccg gactctctgg cggaggcgcc cccccccccg ccacggtgat ctccggcaac 13980
gcggaagctc tcacacaggc gctggaacac taccgggacc aaggcgtcga cgcgaaacgg 14040
atcccggtcg actacgcctc ccactgcccc cacatccagg ccgtggaaca ggaactgtca 14100
cggctgttgc ggggcatcac cccacgggcc gccaccaccc ccttctactc caccaccgac 14160
aaccaatgga ccgacaccac caccctcaac gcccactact ggtaccgaaa cctccgccaa 14220
cccgtccacc tcgccgacgc catcaccaac ctcacccacc aaggccacca caccttcatc 14280
gaaatcagcc cccaccccac cctcaccccc gccatccaag aaaccaccga caccacccac 14340
acccccacca ccgtcatcag cacactccgc cgcaaccaca acgacaccca ccaaatcctc 14400
cacgccctcg cccacgccca caccaccggc caccccatca actggcacac cacccaccaa 14460
caccacaccc caacccccca acacatcgac ctacccacct accccttcca acaccaccac 14520
tactggctca acacccccac ccagacaggg gatgcggcgg ccgtcggcct ggacccggca 14580
catcacccgt tgctgggcgc ggcggtcgcg gtggccgagg gggagggcta tctgctcacc 14640
ggtcggctcg ccctgtccac ccacccctgg ctcgccgatc acaccatcgc cggcgcggtt 14700
gtcctccctg gaactgccct tctcgagatc gcccttcagg cgggccatcg tgtggactgc 14760
tggcgcatcg aagaactcac cctccaatca ccgctgttca tcccggaaga gggagcagta 14820
caggtgcagg catgggtggc ggcaccggat gagaacgggt gccgaagcct gacggtgtcc 14880
tcccgacgcg agggtacgta cgaggacgcc acgtgggtgc gccatgccac gggccgggtc 14940
ggccccgcac cggccgacca ggatgaagcc atcgcacggc tcaccgaccc acaaggcgac 15000
ggagcggcgg cggcggtctg gccaccgcag ggcgctgtcg cgttcaccgc agacgatctg 15060
gagggcctgt acgacgggta cgcggcgcgg ggattcgagt acggcccggt gttccgaggc 15120
ctgcgggcgg cctggcgacg tggcgaggac atcttcgccg aggtgcgcct tcccgacacg 15180
gcggacggcg acgcctccca gttctccgta caccccgccc tgctggacgc cgcactgcac 15240
gccgcggcct tccgcccggc cgacaaactc ccgcacggcg ccctgccgtt ctccttcagc 15300
ggggtgaggc tgcacgggcc cggagcgtcg accctgcggg tgcgcctcac cccggacggc 15360
caggcgcggg acacgcacgc atggtcggtc gcggtggtcg acggcgaggg gcggccggtg 15420
gcctcgatcg catcgctcgc ggtccgcccg gtgtcgacgc aggagttgct ggcggcctcc 15480
ggtacggcgc ggcgggactc gctcttcgcg gtcgagtggg tgaccgccct ggcgccgacc 15540
tcgtcgtccg ttccgcaacg cctggccacg gtggggccca gcgaccgcct cccctcggca 15600
gacgcgtacg cgaacctcgc cgacctggcc gccgcagtgc tggaggcggg ggccccggcg 15660
cccgatgcgg tcgtggtcga ctgcggccgc cgcgatgcgc gcgccaccgc cgtgccggag 15720
gacgtaagga ccctcacccg gcgcatcctg ggtctgctgc aggagtggct ggcggacgag 15780
aggccggcct cgagccggat ggtcgtactg acccgtggtg cggtggccac cactccgggg 15840
gaggacgtgg cggacctggc gggcgcggcg gtgtgcggca tggtgcgctc cgcgcagtcg 15900
gaacatcccg gccggttcgt cctgctggac ctcgaccccg acccggacct cgacggcggg 15960
gaagtgccac cgaccgtcgt tccggcggct ctcgcctgtg gtgagccgca gatcgcggtg 16020
cgtgcgaacc ggcacctggt gccccggctg acccgcgttc cggcgtccgt ccccgtcccc 16080
gggcgtgttc ccgttcccgc cgccgaggca gccgacccgg acaccacgcc cacggcgttc 16140
gaccccgacg gcaccgtagt gatcaccggc ggcaccggca cccttggcgc gatgctcgcg 16200
cgccatctgg tcagccgtca cggtgtacga cacctcctgc tggcatcgcg acgcggaccc 16260
gacgcacccg gcgccaccga gctgcgggcg gaactggccg agctcggcgc cgaggtgacg 16320
gtgcgcgctt gtgacaccgg tgaccgaggc gcgctggcgg atctcatcgc ggggattccc 16380
accggccacc ctttgaccgg tgtggtccac gctgcgggcg tcctggacga cgccaccgtc 16440
gcctcgctca ccccccgaca cctggacacc gcgctgacac ccaaggccga cgccgccttc 16500
catctgcacg agctcacccg ccacgcccgg ccgcgcgcct tcgtcctgtt ctcctcggcc 16560
gccggtgtcc tcggcgcagc cgggcagggc aactatgcgg ccgccaacgc tttcctcgac 16620
gccctcgccg aacaccgcag ggcgcagggc ctgccggcct tgtcgctcgc gtggggcctg 16680
tgggagcagg gcagcggcat gaccgggcat ctcgaccgca ccgaccgggc ccgcatcaac 16740
cgctccggac tcgcccccct cgccacggag gacgctctcg cgctcttcga cgccgccctc 16800
gccggcgatc ggccgttcct ggtgcccgcc cggctggacc tgcggggttc aagcgccgcc 16860
gagaccccgg cgccgctgtt ctccaggatc gccccggctc gtacgacccg gggccggtcc 16920
cccggcgccg agggcgccgc tgaccttcgt acccgtctcg cggcccagga cgccgccgag 16980
cagcgcgaca cgcttctcac gatcgtccgc acccacaccg ccgccgtcct ggggcatgac 17040
acggctgccg ccgtgcggcc ggacggggcc ttccgtgaac tgggtttcga ctccctcgcc 17100
gccgtggaac tccgtaaccg ccttcaaacg accaccgccc tcaccctgcc cgcgaccacc 17160
gtcttcgacc accccacccc cgctgccctc gccgatcatc tgcgtactca gctctgccag 17220
gacgctcagt cctcggcggc ggccacggcc atggcggcga tggcggagct ggccaggctg 17280
gagtccgccg tctccgattc ggtggcgctc gacgacgaca cgcgcagcgg cctcgcggag 17340
cgcctgcggt ccctcgcccg caagatgagc agtggccgtg tcgtcgacca cgacggcggc 17400
ggcgctgcgg acctggatct tcagtcggtc acggacgatg agatgttcga gctgatcgac 17460
aaggaggtca gccgagactg a 17481
<210> 78
<211> 5826
<212> PRT
<213> Artificial Sequence
<220>
<223> milA3 protein of Streptomyces bingchenggensis
<400> 78
Met Ala Ala Gly His Asp Lys Val Ile Glu Ala Leu Arg Ala Ser Leu
1 5 10 15
Lys Thr Asn Glu Arg Gln Arg Glu Gln Ile His Arg Leu Thr Thr Ala
20 25 30
Ala Arg Glu Pro Ile Ala Ile Ile Gly Met Ala Cys Arg Tyr Pro Gly
35 40 45
Gly Val Gly Ser Pro Glu Asp Leu Trp Glu Leu Val Ala Ala Gly Arg
50 55 60
Asp Ala Ile Gly Thr Phe Pro Glu Asp Arg Gly Trp Asp Val Glu Arg
65 70 75 80
Leu Tyr Asp Pro Asp Pro Glu Arg Ala Gly Thr Ser Cys Thr Gln His
85 90 95
Gly Gly Phe Leu Tyr Gln Ala Gly Glu Phe Asp Pro Gly Phe Phe Gly
100 105 110
Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu
115 120 125
Leu Glu Ile Ser Trp Glu Val Phe Glu Arg Ala Gly Ile Asp Pro Ala
130 135 140
Ser Val Arg Gly Ser Arg Thr Gly Val Phe Ala Gly Val Met Tyr His
145 150 155 160
Asp Tyr Gly Ser Arg Leu His Thr Val Pro Glu Gly Phe Glu Gly Tyr
165 170 175
Val Gly Asn Gly Ser Gly Gly Gly Val Ala Ser Gly Arg Val Ala Tyr
180 185 190
Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser
195 200 205
Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ala Gly
210 215 220
Glu Cys Ser Leu Ala Leu Ala Gly Gly Val Thr Val Met Ser Thr Pro
225 230 235 240
Ser Leu Phe Val Glu Tyr Ser Arg Gln Arg Ala Leu Ala Ala Asp Gly
245 250 255
Arg Cys Lys Ala Tyr Gly Ala Gly Ala Asp Gly Thr Gly Trp Ala Glu
260 265 270
Gly Ala Gly Met Leu Leu Val Glu Arg Leu Thr Asp Ala Gln Arg Leu
275 280 285
Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp
290 295 300
Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala Gln Gln Arg
305 310 315 320
Ala Ile Arg Gln Ala Leu Ala Ser Ala Gly Val Ser Ala Ser Glu Val
325 330 335
Asp Ala Val Glu Gly His Gly Thr Gly Thr Arg Leu Gly Asp Pro Ile
340 345 350
Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Gln Arg Pro Ala Asp
355 360 365
Arg Pro Leu Trp Leu Gly Ser Met Lys Ser Asn Val Gly His Ala Gln
370 375 380
Ala Ala Ala Gly Val Gly Gly Ile Ile Lys Met Val Met Ala Met Arg
385 390 395 400
Ser Gly Thr Leu Pro Arg Thr Leu His Ala Asp Glu Pro Ser Pro His
405 410 415
Ile Asp Trp Asp Ser Gly Ala Val Arg Leu Leu Thr Glu Pro Val Ala
420 425 430
Trp Pro Glu Arg Asp Arg Pro Arg Arg Ala Ala Val Ser Ser Phe Gly
435 440 445
Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Ala Ala Ser Gln Thr
450 455 460
Ala Pro Gln Thr Asp Ser Ala Ser Gln Ala Glu Thr Asp Asp Ala Pro
465 470 475 480
Ala Pro His Gly Ala Pro Gly His Ala Val Ala Gly Pro Leu Leu Trp
485 490 495
Pro Leu Ser Gly Ala Thr Ala Glu Ala Leu Arg Ala Gln Ala Gly Glu
500 505 510
Leu Arg Arg Phe Val Ala Ala Asp Glu Leu Leu Arg Pro Ala Asp Val
515 520 525
Gly His Thr Leu Val Phe Gly Arg Ser Asp Leu Ala His Arg Ala Val
530 535 540
Val Leu Gly Ser Asp Arg Glu Thr Leu Leu Arg Ala Leu Asp Thr Leu
545 550 555 560
Ala Gly Glu Gly Pro Asp Asp Gly Ser Val Val Arg Gly Met Ala Ala
565 570 575
Ala Gly Ala Gly Ala Gly Val Val Phe Val Phe Pro Gly Gln Gly Gly
580 585 590
Gln Trp Ala Gly Met Gly Leu Arg Leu Leu Glu Thr Ser Ser Phe Phe
595 600 605
Ala Glu Arg Met Ala Glu Cys Glu Ala Ala Leu Ala Pro Tyr Ala Asp
610 615 620
Trp Ser Leu Leu Asp Val Leu Arg Arg Asp Pro Gly Asp Pro Val Trp
625 630 635 640
Glu Arg Ala Asp Val Val Gln Pro Met Leu Phe Ser Val Met Val Ser
645 650 655
Leu Ala Gln Leu Trp Arg Ser Tyr Gly Val Glu Pro Asp Ala Val Leu
660 665 670
Gly His Ser Gln Gly Glu Ile Ala Ala Ala His Ile Cys Gly Ala Leu
675 680 685
Thr Leu Asp Asp Ala Ala Lys Val Val Ala Leu Arg Ser Arg Ala Leu
690 695 700
Gln Thr Leu Arg Gly Ser Gly Gly Met Ala Ser Val Pro Leu Pro Ala
705 710 715 720
Asp Glu Val Thr Gly Leu Leu Arg Thr Asp Ser Leu Trp Val Ala Ala
725 730 735
Val Asn Ala Pro Thr Ala Thr Val Ile Ser Gly Asp Ala Asp Ser Leu
740 745 750
Ala Glu Ala Leu Glu His Tyr Arg Asp Gln Gly Val Glu Ala Lys Arg
755 760 765
Val Pro Val Asp Tyr Ala Ser His Cys Pro His Ile Glu Ala Val Glu
770 775 780
Gln Glu Leu Leu Gly Leu Leu Arg Gly Ile Ala Pro Arg Ala Ala Asp
785 790 795 800
Ile Pro Phe Tyr Ser Thr Val Asp Asn Gln Trp Ala Asp Thr Met Gly
805 810 815
Leu Asp Ala Arg Tyr Trp Tyr Arg Asn Leu Arg Arg Pro Val Arg Phe
820 825 830
Ala Glu Ala Leu Arg Ala Leu Gly Ala Ala Glu Tyr Arg Thr Tyr Val
835 840 845
Glu Val Gly Pro His Pro Thr Leu Thr Pro Ala Ile Glu Asp Thr Thr
850 855 860
Glu Ala Ala Gly Val Ala Ala Thr Val Val Gly Ser Leu Arg Arg Gly
865 870 875 880
Glu Asp Asp Ala His Arg Ile Leu Thr Ser Leu Ala Arg Ala His Ile
885 890 895
His Gly Leu Pro Val Ala Trp Asp Arg His Tyr Arg Ala Leu Ala Pro
900 905 910
Glu Ala Asn His Val Asp Leu Pro Thr Tyr Ala Phe Gln Arg Arg Arg
915 920 925
Tyr Trp Leu Asp Ala Pro Ala Thr Thr Gly Asp Val Thr Ala Ala Gly
930 935 940
Leu Ala Pro Val Gly His Pro Leu Leu Gly Ala Ala Val Gly Leu Ala
945 950 955 960
Glu Gly Asp Gly Tyr Leu Leu Thr Gly Arg Leu Ala Pro His Thr His
965 970 975
Pro Trp Leu Thr Asp His Ala Val Ala Gly Thr Val Leu Leu Pro Gly
980 985 990
Thr Ala Tyr Val Glu Leu Ala Val His Val Gly Gly His Leu Gly Cys
995 1000 1005
Pro Arg Leu Glu Glu Leu Thr Leu His Ala Pro Leu Val Leu Pro Asp
1010 1015 1020
Thr Gly Gly Val Ala Leu Gln Val Ala Val Gly Ala Pro Asp Glu Thr
1025 1030 1035 1040
Gly Arg Arg Ala Leu Ser Val Tyr Ala Gln Arg Asp Asp Asp Pro Ala
1045 1050 1055
Trp Glu Gly Ala Ala Arg Gly Ala Trp Thr Arg His Ala Thr Gly Thr
1060 1065 1070
Leu Ala Ala Glu Ala Pro Thr Asp Gly Ile Ser Gly Ala Asp Gly Ala
1075 1080 1085
Gly Thr Leu Ala Gly Ala Trp Pro Pro Pro Gly Ala Glu Pro Leu Asp
1090 1095 1100
Ile Ser Gly Leu Tyr Asp Thr Leu Ala Ala Ala Asp Phe Gly Tyr Gly
1105 1110 1115 1120
Pro Ala Phe Gln Gly Leu Arg Ala Val Trp Arg Gln Gly Glu Glu Thr
1125 1130 1135
Tyr Ala Glu Val Arg Leu Pro Asp Gln Val Ala Ala Asp Ala Pro Arg
1140 1145 1150
Phe Cys Leu His Pro Ala Leu Leu Asp Ala Ala Leu His Pro Leu Ala
1155 1160 1165
Leu Asp Ser Gly Arg Ser Glu Glu Asn Pro Ala Gly His Gly Leu Leu
1170 1175 1180
Pro Phe Ala Trp Arg Gly Val Ser Leu Arg Ser Pro Gly Thr Pro Thr
1185 1190 1195 1200
Leu Arg Val Arg Leu Arg Pro Gln Gly Pro Asp Ser Ile Ala Val Asp
1205 1210 1215
Val Ala Asp Glu Thr Gly Ala Pro Val Ala Ser Ala Glu Ser Leu Thr
1220 1225 1230
Leu Arg Pro Val Ala Leu Glu Asp Leu Arg Ala Leu Gly Gly Gln Ala
1235 1240 1245
Gly Asp Thr Leu Tyr Ala Leu Glu Trp Thr Ala Ala Pro Glu Pro Pro
1250 1255 1260
Ala Thr Ala Leu Gly Arg Cys Ala Val Ile Gly Gln Ala Ile Pro Gly
1265 1270 1275 1280
Trp Ala Ala Ala Leu Glu Thr Ala Ala Ala Gly Pro Val Arg Arg Tyr
1285 1290 1295
Pro Asp Leu Ala Gly Leu Val Thr Ala Leu Asp Ala Gly Asp Pro Pro
1300 1305 1310
Pro Asp Leu Val Phe Val Gly Cys Pro Pro Ala Ala Ala Gly Pro Asp
1315 1320 1325
Asp Thr Thr Val Ala Asp Val His Thr Ala Arg Thr Arg Val Arg Thr
1330 1335 1340
Arg Gln Ala Leu Asp Leu Leu Gln Gly Trp Leu Gly Glu Ala Arg Leu
1345 1350 1355 1360
Ala Gly Ala Arg Leu Val Leu Val Thr Cys Gly Ala Val Ala Thr Gly
1365 1370 1375
Pro Ala Glu Gly Val Met Asp Leu Ala Gly Ala Ala Ile Cys Gly Leu
1380 1385 1390
Val Arg Ser Ala Gln Ala Glu Glu Pro Asp Arg Ile Leu Leu Val Asp
1395 1400 1405
Leu Asp Ala Ala Glu Glu Ser Trp Ala Ala Leu Pro Arg Ala Val Ala
1410 1415 1420
Leu Gly Glu Pro Gln Met Ala Ile Arg Ala Gly Gln Pro His Met Ala
1425 1430 1435 1440
Arg Leu Val Arg Ala Asp Thr Glu Gly Gly Ala Leu Leu Thr Pro Pro
1445 1450 1455
Gln Gly Ser Gly Gly Trp Arg Leu Asp Cys Ala Asp Ala Gly Thr Val
1460 1465 1470
Gln Gly Leu Ala Pro Val Ala Ser Ser Ala Asp Arg Asp Pro Leu Gly
1475 1480 1485
Pro His Gln Val Arg Ile Glu Val Arg Ala Ala Gly Leu Asn Phe Arg
1490 1495 1500
Asp Val Leu Val Ala Leu Gly Met Val Pro Gly Gln Arg Gly Leu Gly
1505 1510 1515 1520
Ser Glu Gly Ala Gly Val Val Leu Glu Ala Gly Pro Glu Val Ala Asp
1525 1530 1535
Leu Ala Pro Gly Asp Arg Val Met Gly Val Phe Ala Asp Ala Phe Gly
1540 1545 1550
Pro Phe Ala Ile Ala Asp Arg Ala Thr Val Ile Arg Val Pro Asp His
1555 1560 1565
Trp Thr Phe Gly Gln Ala Ala Ala Val Pro Val Val Phe Ala Thr Ala
1570 1575 1580
Tyr Tyr Gly Leu Val Asp Leu Ala Gly Leu Arg Pro Gly Glu Ser Val
1585 1590 1595 1600
Leu Val His Ala Ala Ala Gly Gly Val Gly Leu Ala Ala Val Gln Leu
1605 1610 1615
Ala Arg His Leu Gly Ala Glu Val Tyr Ala Thr Ala Ser Pro Gly Lys
1620 1625 1630
Trp Asp Thr Leu Arg Ala His Gly Ile Pro Pro Glu Arg Ile Ala Ser
1635 1640 1645
Ser Arg Thr Leu Asp Phe Glu Ser Arg Phe Thr Gly Arg Asn Ile Asp
1650 1655 1660
Val Val Leu Asn Ser Leu Ala His Glu Tyr Val Asp Ala Ser Leu Arg
1665 1670 1675 1680
Leu Val Ser Gly Asp Ser Gly Arg Phe Leu Glu Met Gly Lys Thr Asp
1685 1690 1695
Leu Arg Asp Pro Glu Glu Val Ala Gln Ala Tyr Pro Gly Val Ala Tyr
1700 1705 1710
Arg Ala Tyr Asp Leu Met Glu Ala Gly Pro Glu Arg Ile Gly Glu Ile
1715 1720 1725
Leu Arg Thr Val Leu Arg Leu Phe Asp Glu Gly Val Leu Thr Pro Leu
1730 1735 1740
Pro Leu Thr Cys Trp Asp Ile Arg Gln Ala Arg Asp Ala Phe Arg Gln
1745 1750 1755 1760
Leu Gln Gln Gly Arg Thr Val Gly Lys Asn Val Leu Thr Leu Asp Arg
1765 1770 1775
Thr Pro Asp Pro Asp Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Thr
1780 1785 1790
Leu Gly Ala Ala Leu Ala Arg His Leu Ala Ala Thr Gly Arg Ala Arg
1795 1800 1805
His Leu Leu Leu Ile Ser Arg Arg Gly Leu Asp Ala Pro Gly Ala Pro
1810 1815 1820
Glu Leu Ile Ala Glu Ile Asp Glu Leu Gly Ala Thr Ala Thr Val Ala
1825 1830 1835 1840
Thr Cys Asp Val Gly Asp Arg Ala Ala Leu Ala Glu Leu Leu Gly Arg
1845 1850 1855
Ile Pro Ala Glu His Pro Leu Thr Ala Val Val His Ala Ala Gly Thr
1860 1865 1870
Leu Asp Asp Ala Thr Leu Gly Ser Leu Thr Ala Arg His Leu Asp Thr
1875 1880 1885
Val Leu Pro Ala Lys Ala Asp Ala Ala Trp His Leu His Asp Leu Thr
1890 1895 1900
Cys Arg Leu Asp Leu Ala Ala Phe Val Leu Phe Ser Ser Ala Ala Gly
1905 1910 1915 1920
Val Leu Gly Ser Pro Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Phe
1925 1930 1935
Leu Asp Ala Leu Ala Phe Gln Arg Arg Ala Met Gly Leu Pro Ala Val
1940 1945 1950
Ser Leu Ala Trp Gly Leu Trp Glu Glu Ala Ser Gly Met Thr Gly His
1955 1960 1965
Leu Asp Gln Thr Asp Arg Thr Arg Met Ala Arg Val Gly Leu Arg Pro
1970 1975 1980
Leu Ala Thr Asp Glu Ala Leu Ala Leu Phe Asp Asn Ala Leu Val Asp
1985 1990 1995 2000
Gly Pro Pro Leu Leu Leu Pro Ala Arg Ile Asp Thr Lys Ala Leu Arg
2005 2010 2015
Gly Thr Thr Ala Pro Pro Leu Phe Gln Ser Leu Val Arg Pro Thr Thr
2020 2025 2030
Gly His Arg Pro Arg Pro Ala Thr Pro Asp Gly Arg Ser Ser Leu Arg
2035 2040 2045
Ala Arg Leu Ala Gly Leu Asp Pro Ala Ala Gln His Glu Val Leu Leu
2050 2055 2060
Thr Leu Val Arg Gly His Ala Ala Thr Val Leu Gly His Pro Ser Pro
2065 2070 2075 2080
Asp Ala Ile Ala Arg Glu Ala Ala Phe Arg Asp Leu Gly Phe Asp Ser
2085 2090 2095
Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Lys Glu Ala Thr Gly Leu
2100 2105 2110
Arg Leu Pro Pro Pro Pro Arg Leu Lys Glu Ala Thr Gly Leu Arg Leu
2115 2120 2125
Pro Ala Thr Ile Val Phe Asp His Pro Thr Pro Ala Ala Leu Ala Gln
2130 2135 2140
His Leu Arg Asp Gly Leu Ile Gly Gly Ala Asp Thr Val Thr Leu Ala
2145 2150 2155 2160
Ala Ala Pro Ala Pro Ser Lys Val Ala Met Val Ala Asp Glu Ala Ile
2165 2170 2175
Ala Ile Ile Gly Met Ala Cys Arg Tyr Pro Gly Gly Val Arg Ser Ala
2180 2185 2190
Glu Gly Leu Trp Asp Leu Val Ala Ser Gly Thr Asp Ala Met Ser Gly
2195 2200 2205
Phe Pro Ser Asp Arg Gly Trp Asp Leu Asp Arg Leu Tyr Ala Pro Gln
2210 2215 2220
Asp Gln Asp Val Pro Gly Thr Thr Tyr Thr Arg His Gly Gly Phe Leu
2225 2230 2235 2240
His Asp Ala Gly Lys Phe Asp Ala Gly Phe Phe Gly Ile Gly Pro Arg
2245 2250 2255
Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Thr Ser
2260 2265 2270
Trp Glu Val Phe Glu His Ala Gly Ile Asp Pro Ser Ser Val Arg Arg
2275 2280 2285
Ser Arg Thr Gly Val Phe Ala Gly Val Met Pro Thr Asp Tyr Gly Pro
2290 2295 2300
Arg Leu Gln Asp Thr Val Ala Glu Val Glu Gly Tyr Val Leu Thr Gly
2305 2310 2315 2320
Asn Ser Gly Ser Val Ala Ser Gly Arg Ile Ala Tyr Thr Phe Gly Leu
2325 2330 2335
Glu Gly Pro Ala Val Ser Val Asp Thr Ala Cys Ser Ser Ser Leu Val
2340 2345 2350
Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ala Gly Glu Cys Ser Met
2355 2360 2365
Ala Leu Ala Gly Gly Val Thr Val Met Ala Thr Pro Gly Ala Phe Val
2370 2375 2380
Glu Phe Ala Arg Gln Arg Gly Leu Ser Val Asp Gly Arg Cys Lys Ala
2385 2390 2395 2400
Phe Gly Val Gly Ala Asp Gly Thr Gly Trp Ala Glu Gly Val Gly Met
2405 2410 2415
Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu Gly His Arg Val
2420 2425 2430
Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn
2435 2440 2445
Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln Arg Val Ile Arg Gln
2450 2455 2460
Ala Leu Ala Ser Ala Arg Val Gly Gly Ala Asp Val Asp Val Val Glu
2465 2470 2475 2480
Gly His Gly Thr Gly Thr Arg Leu Gly Asp Pro Ile Glu Ala Gln Ala
2485 2490 2495
Leu Leu Ala Thr Tyr Gly Gln Glu Arg Pro Asp Asp Arg Pro Val Trp
2500 2505 2510
Leu Gly Ser Val Lys Ser Asn Ile Gly His Ala Gln Ala Ala Ala Gly
2515 2520 2525
Val Ala Gly Val Ile Lys Met Val Met Ala Met Arg Tyr Gly Val Leu
2530 2535 2540
Pro Arg Thr Leu His Val Gln Glu Pro Ser Pro His Val Asp Trp Ser
2545 2550 2555 2560
Ser Gly Gly Val Arg Leu Leu Thr Glu Ala Val Pro Trp Pro Glu Thr
2565 2570 2575
Gly Arg Ala Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr
2580 2585 2590
Asn Ala His Ile Ile Leu Glu Gln Ala Pro Pro Glu Glu His Asp Asp
2595 2600 2605
Pro Ala Asp Val Ser Ser Gly Ser Phe Pro Trp Met Val Ser Ala Lys
2610 2615 2620
Ser Glu Gln Ala Leu Gln Ala Gln Ala Ala Gln Leu Arg Ala Tyr Leu
2625 2630 2635 2640
Ala Ala His Pro Glu Leu Gly Leu Ala Asp Val Gly Tyr Ala Leu Ala
2645 2650 2655
Ser Gly Arg Thr Ala Phe Gly His Arg Ala Val Leu Leu Gly Pro Asp
2660 2665 2670
Arg Glu Ala Phe Val Glu Glu Leu Gly Ala Leu Glu Ala Gly Glu Glu
2675 2680 2685
His Ala Gly Leu Val Arg Gly Val Ala Thr Gly Ala Gly Lys Leu Ala
2690 2695 2700
Phe Val Cys Ser Gly Gln Gly Thr Gln Arg Pro Arg Met Gly His Gly
2705 2710 2715 2720
Leu Tyr Ser Pro Ser Arg Cys Ser Pro Gln Pro Trp Thr Lys Pro Ala
2725 2730 2735
His Thr Trp Thr His Thr Ser Thr Ile Pro Cys Gly Met Ser Cys Ser
2740 2745 2750
Pro Ser Arg Ala Pro Thr Pro Pro Ser Cys Ser Thr Arg Pro Ala Thr
2755 2760 2765
Pro Ser Pro Arg Cys Ser Pro Ser Arg Ser Pro Cys Thr Ala Trp Ser
2770 2775 2780
Pro Asn Thr Thr Ala Leu Pro Pro Thr Thr Thr Pro Ala Ile Pro Ser
2785 2790 2795 2800
Glu Arg Ser Pro Arg Pro Thr Ser Pro Gly Ser Ser Pro Ser Pro Thr
2805 2810 2815
Arg Pro Ala Trp Ser Pro Pro Ala Pro Ala Ser Cys Asn Leu Ser Pro
2820 2825 2830
Pro Pro Ala Pro Met Thr Thr Leu Gln Ala Asp Pro Asp Glu Leu His
2835 2840 2845
Glu His Leu Thr Arg Cys Glu Gly Arg Val Ser Leu Ala Ala Val Asn
2850 2855 2860
Ala Pro Gly Ser Val Val Ile Ser Gly Asp Arg His Asp Val Asp Ala
2865 2870 2875 2880
Thr Ala Glu Asn Leu Arg Ala Met Gly Arg Lys Thr Thr Ala Leu Lys
2885 2890 2895
Val Ser Gly Ala Phe His Ser His His Ile Asp Pro Leu Leu Asn Glu
2900 2905 2910
Leu Arg Asn Thr Ala Glu Thr Leu Thr Tyr His Pro Pro His Thr Pro
2915 2920 2925
Leu Ile Thr Thr Asn Pro Thr Asp His Asp Pro Thr Thr Pro His Tyr
2930 2935 2940
Trp Val Arg Gln Ala Arg Glu Thr Val His Tyr Ala His Thr Thr Gln
2945 2950 2955 2960
Gln Leu His Thr His Gly Val Thr Ala Tyr Leu Glu Leu Gly Pro Asp
2965 2970 2975
His Thr Leu Thr Ala Leu Thr His His Asn Leu Pro Asp His Thr Pro
2980 2985 2990
Leu Ala Val Pro Leu Leu His Pro Asp Gln Ser Glu Thr His Thr Thr
2995 3000 3005
His Thr Ala Leu Ala His Leu His Thr His Gly His Pro Thr Thr Trp
3010 3015 3020
His His His His Thr Pro Thr His Tyr His Pro Asn Leu Pro Thr Tyr
3025 3030 3035 3040
Pro Phe Gln His His His Tyr Trp Leu Asn Thr Thr Thr Ala Thr Gly
3045 3050 3055
Asp Met Ser Ala Ala Gly Leu Glu Pro Ala Arg His Pro Leu Leu Gly
3060 3065 3070
Ala Ala Val Gly Leu Ala Asp Gly Glu Gly Leu Leu Phe Thr Gly Arg
3075 3080 3085
Ile Ser Leu Arg Thr His Pro Trp Leu Ala Asp His Ala Val Gly Gly
3090 3095 3100
Ala Val Leu Leu Pro Gly Thr Ala Phe Leu Glu Leu Ala Leu Gln Ala
3105 3110 3115 3120
Ala Ala His Ala Asp Cys Arg Arg Val Glu Glu Leu Thr Leu His Thr
3125 3130 3135
Pro Leu Val Val Pro Asp Ser Ala Gly Val Val Leu Gln Val Thr Val
3140 3145 3150
Ala Ala Pro Asn Glu Ala Gly Asn Arg Ala Val Asp Ile Tyr Ser Arg
3155 3160 3165
Ile Asp Val Gly Gly Leu Thr Ala Asp Ser Ala Gly Glu Pro Trp Thr
3170 3175 3180
Arg His Ala Ala Gly Tyr Leu Ala Asp Lys Pro Asp Pro Asp Cys Gly
3185 3190 3195 3200
Asp Ser Ala Asp Gly Val Met Pro Ala Gly Ala Trp Pro Pro Pro Gly
3205 3210 3215
Ala Val Ala Val Asp Leu Glu Gly Leu Tyr Glu Gln Leu Ala Glu Gly
3220 3225 3230
Gly Phe His Tyr Gly Ala Ala Phe Arg Cys Leu Asp Ala Ala Trp Gln
3235 3240 3245
Arg Gly Asp Glu Val Phe Ala Thr Ala Tyr Met Ser Glu Asp Gln Leu
3250 3255 3260
Gly Asp Thr Ala Ala Ala Arg Phe Ala Leu His Pro Ala Leu Leu Asp
3265 3270 3275 3280
Ser Ala Leu His Thr Ile Pro Leu Leu Pro Ser Leu Arg Gly Gln Gln
3285 3290 3295
Asp Ser Gly Leu Pro Phe Thr Trp Thr Gly Val Thr Leu Arg Ala Ser
3300 3305 3310
Gly Ala Thr Ala Leu Arg Val Arg Leu Arg Pro Asp Gly His Gly Pro
3315 3320 3325
Gly Ala Val Ser Val Asp Val Ser Asp Glu Ala Gly Glu Pro Val Ala
3330 3335 3340
Ser Val Arg Ser Leu Ala Leu Arg Pro Val Thr Arg Ala Glu Leu His
3345 3350 3355 3360
Thr Ala Glu Leu Arg Thr Ala Ala Pro Val Ala Pro His Gly Ser Leu
3365 3370 3375
Phe Glu Val Arg Trp Glu Pro Val Pro Gln Pro Ser Ala Ala Glu Glu
3380 3385 3390
Ala Ala Pro Trp Val Met Ile Gly Thr Gly Pro Thr Leu Arg Pro Val
3395 3400 3405
Glu Asp Phe Val Thr Pro Pro Glu Arg Thr Tyr Ala Asp Leu Ala Ala
3410 3415 3420
Leu Cys Val Ala Ile Ala Asp Asp Ala Pro Val Pro Arg Thr Val Val
3425 3430 3435 3440
Ala Trp Ser Pro Ala Gly Ser Glu Asp Glu Ser Ser Glu Ala Leu Arg
3445 3450 3455
Gln Ala Thr His His Met Leu Gly Leu Leu Gln Gln Trp Leu Ala Asp
3460 3465 3470
Ser Arg Phe Ala Asp Ser Arg Leu Val Ile Leu Thr Arg Ala Ala Val
3475 3480 3485
Ala Thr Ala Pro Asp Glu Glu Val Glu Asp Leu Ala Gly Ala Ala Ala
3490 3495 3500
Arg Gly Leu Ile Arg Ser Ala Gln Ser Glu His Pro Asp Arg Phe Val
3505 3510 3515 3520
Leu Leu Asp Leu Asp Asp Arg Pro Ala Asp Ala Lys Asp His Asp Arg
3525 3530 3535
Met Leu Ser Met Ala Leu Ala Cys Gly Glu Pro Glu Val Ala Val Arg
3540 3545 3550
Asp Gly Ala Leu Arg Thr Pro Arg Leu Ser Pro Leu Ala Gly Thr Ala
3555 3560 3565
Thr Glu Ala Met Asp Glu His Pro Trp Asp Gln Asp Gly Thr Val Leu
3570 3575 3580
Ile Thr Gly Gly Thr Gly Ser Leu Gly Ala Met Leu Ala Arg His Leu
3585 3590 3595 3600
Val Ala Thr His Gly Val Arg His Leu Met Leu Ile Ser Arg Arg Gly
3605 3610 3615
Leu Asp Ala Pro Gly Ala Arg Arg Leu Gly Val Glu Leu Ala Glu Leu
3620 3625 3630
Gly Ala Gln Val Thr Ile Thr Ala Cys Asp Ala Ala Asp Gln Arg Gln
3635 3640 3645
Leu Ala Asn Val Leu Ser Glu Ile Ser Val Asp His Pro Leu Thr Ala
3650 3655 3660
Val Val His Ala Ala Gly Val Leu Asp Asp Gly Val Ile Thr Ser Leu
3665 3670 3675 3680
Thr Pro Glu Gly Leu Thr His Val Leu Arg Ala Lys Val Asp Ser Ala
3685 3690 3695
Leu Asn Leu His Gln Leu Thr Arg Asp Leu Pro Leu Ser Ala Phe Val
3700 3705 3710
Leu Phe Ser Ser Leu Ala Gly Val Met Gly Ser Ala Gly Gln Gly Asn
3715 3720 3725
Tyr Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu Ala Ser His Arg Arg
3730 3735 3740
Ala Ala Arg Leu Pro Ala Val Ser Leu Ala Trp Gly Val Trp Glu Gln
3745 3750 3755 3760
Thr Glu Gly Met Thr Gly Gln Leu Glu Ala Thr Asp His Ala Arg Leu
3765 3770 3775
Arg Arg Ser Gly Leu Arg Pro Leu Ala Ile Ser Glu Gly Leu Glu Leu
3780 3785 3790
Phe Asp Lys Ala Leu Ser Cys Gly His Ala Leu Val Val Pro Ala Ala
3795 3800 3805
Leu Ser Thr Arg Glu Leu Gln Thr Ser Gly Ser Val Pro Pro Phe Leu
3810 3815 3820
Arg His Leu Thr Gly Val Ala Pro Ala Arg Pro Ser Arg Thr Arg Asp
3825 3830 3835 3840
Ala Ser Ala Gly Glu Pro Thr Ser Leu Arg Arg Arg Leu Thr Gly Leu
3845 3850 3855
Gly Pro Glu Glu Arg Leu Arg Glu Val Leu Arg Leu Val Arg Ser Arg
3860 3865 3870
Ala Ala Ala Val Leu Gly His Gly Thr Ala Glu Ser Val Pro Ala Asp
3875 3880 3885
Ser Ala Phe Arg Asp Leu Gly Phe Asp Ser Leu Ala Ala Val Asp Leu
3890 3895 3900
Arg Asn Arg Leu Gln Gln Ala Thr Gly Leu Arg Leu Pro Ala Gly Leu
3905 3910 3915 3920
Ile Phe Asp Arg Pro Arg Pro Asp Val Leu Ala Arg Phe Leu Cys Asp
3925 3930 3935
Glu Leu Ala Gly Ala Gly Gly Thr Ser Ala Ala Thr Ala Ala Pro Pro
3940 3945 3950
Val Ala Ala Gly Gly Gly Gly Gly Arg Gly Gly Ala Gly Gly His Arg
3955 3960 3965
Arg His Gly Met Pro Val Ser Gly Arg Cys Ala Val Gly Arg Gly Pro
3970 3975 3980
Val Gly Ser Gly Arg Leu Arg Tyr Gly Arg Val Gly Asp Phe Pro Ala
3985 3990 3995 4000
Asp Arg Gly Trp Glu Val Glu Arg Leu Tyr Asp Pro Asp Pro Asp Arg
4005 4010 4015
Thr Gly Thr Ser Tyr Thr Arg Gln Gly Gly Phe Leu Tyr Asp Ala Gly
4020 4025 4030
Glu Phe Asp Ala Ala Phe Phe Gly Ile Gly Pro Arg Glu Ala Val Ala
4035 4040 4045
Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Ile Ser Trp Glu Ala Leu
4050 4055 4060
Glu Arg Ala Gly Ile Asp Pro Ala Ser Leu Arg Gly Ser Ser Thr Gly
4065 4070 4075 4080
Val Phe Ala Gly Val Met Tyr His Asp Tyr Gly Thr Arg Leu Arg Glu
4085 4090 4095
Ile Pro Glu Gly Tyr Glu Gly Tyr Ile Gly Asn Gly Asn Ala Gly Ser
4100 4105 4110
Val Ala Ser Gly Arg Val Ala Tyr Thr Phe Gly Leu Glu Gly Pro Ala
4115 4120 4125
Val Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu
4130 4135 4140
Ala Cys Gln Ala Leu Arg Ser Gly Glu Cys Ser Met Ala Leu Ala Gly
4145 4150 4155 4160
Gly Val Thr Val Met Ser Thr Pro Thr Thr Phe Val Glu Phe Ser Arg
4165 4170 4175
Gln Arg Gly Leu Ala Pro Asp Gly Arg Cys Lys Ser Phe Gly Ala Gly
4180 4185 4190
Ala Asp Gly Thr Gly Trp Ala Glu Gly Ala Gly Met Leu Leu Val Glu
4195 4200 4205
Arg Leu Ser Asp Ala Arg Arg Asn Gly His Arg Val Leu Ala Val Val
4210 4215 4220
Arg Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala
4225 4230 4235 4240
Pro Asn Gly Pro Ser Gln Glu Arg Val Ile Arg Gln Ala Trp Ala Asn
4245 4250 4255
Ala Gly Val Ala Ala Met Asp Ile Asp Ala Val Glu Gly His Gly Thr
4260 4265 4270
Gly Thr Thr Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu Leu Gly Thr
4275 4280 4285
Tyr Gly Gln Gly Arg Ser Ala Asp Arg Pro Leu Trp Leu Gly Ser Ile
4290 4295 4300
Lys Ser Asn Val Gly His Thr Gln Ala Ala Ala Gly Val Gly Gly Val
4305 4310 4315 4320
Ile Lys Met Val Met Ala Met Arg His Gly Leu Leu Pro Gln Thr Leu
4325 4330 4335
His Ala Glu Glu Pro Ser Pro His Val Asp Trp Ser Gly Gly Thr Val
4340 4345 4350
Arg Leu Leu Thr Glu Ser Val Ala Trp Pro Glu Gln Gly Arg Met Arg
4355 4360 4365
Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val
4370 4375 4380
Ile Leu Glu Gln Ala Pro Pro Ala Ala Glu Thr His Glu Pro Ala Glu
4385 4390 4395 4400
Pro Asn Thr Ala Pro Gly Pro Leu Pro Trp Ala Ile Ser Ala Lys Ser
4405 4410 4415
Pro Gln Ala Leu Arg Ala Gln Ala Arg Gln Leu His Thr Tyr Leu Thr
4420 4425 4430
Asn Ala Pro Glu Ala Asn Pro Ala Asp Val Gly His Thr Leu Ala Thr
4435 4440 4445
Gly Arg Ala Ser Phe Glu His Arg Ala Val Val Ile Gly Ser Asp Arg
4450 4455 4460
Ala Glu Phe Leu Gly Gly Leu Asp Ala Leu Ala Ala Asp Glu Ala His
4465 4470 4475 4480
Thr Ala Val Val Thr Gly Ile Ala Arg Lys Ala Gly Asp Gln Gly Lys
4485 4490 4495
Val Val Phe Val Phe Pro Gly Gln Gly Gly Gln Trp Ala Gly Met Gly
4500 4505 4510
Leu Arg Leu Leu Lys Thr Ser Pro Val Phe Ala Gln Ser Ile Gln Ala
4515 4520 4525
Cys Glu Gln Ala Leu Ala Pro His Thr Asp Trp Thr Leu Thr Asp Ile
4530 4535 4540
Leu His Arg Pro His Thr Asp Pro Leu Trp Gln Arg Ala Asp Val Ile
4545 4550 4555 4560
Gln Pro Val Leu Phe Ala Leu Met Thr Ser Leu Ala Ala Leu Trp Gln
4565 4570 4575
Ser His Gly Leu Asn Pro Asp Ala Val Ile Gly His Ser Gln Gly Glu
4580 4585 4590
Ile Thr Ala Ala His Ile Ser Gly Ala Leu Ser Leu Glu Asp Ala Ala
4595 4600 4605
Lys Thr Val Ala Leu Arg Ser Arg Ala Leu Gln Thr Leu Arg Gly Ser
4610 4615 4620
Gly Gly Met Ala Ser Val Pro Leu Pro Ala Asp Glu Val Thr Gly Leu
4625 4630 4635 4640
Leu Arg Thr Gly Leu Ser Gly Gly Gly Ala Pro Pro Pro Ala Thr Val
4645 4650 4655
Ile Ser Gly Asn Ala Glu Ala Leu Thr Gln Ala Leu Glu His Tyr Arg
4660 4665 4670
Asp Gln Gly Val Asp Ala Lys Arg Ile Pro Val Asp Tyr Ala Ser His
4675 4680 4685
Cys Pro His Ile Gln Ala Val Glu Gln Glu Leu Ser Arg Leu Leu Arg
4690 4695 4700
Gly Ile Thr Pro Arg Ala Ala Thr Thr Pro Phe Tyr Ser Thr Thr Asp
4705 4710 4715 4720
Asn Gln Trp Thr Asp Thr Thr Thr Leu Asn Ala His Tyr Trp Tyr Arg
4725 4730 4735
Asn Leu Arg Gln Pro Val His Leu Ala Asp Ala Ile Thr Asn Leu Thr
4740 4745 4750
His Gln Gly His His Thr Phe Ile Glu Ile Ser Pro His Pro Thr Leu
4755 4760 4765
Thr Pro Ala Ile Gln Glu Thr Thr Asp Thr Thr His Thr Pro Thr Thr
4770 4775 4780
Val Ile Ser Thr Leu Arg Arg Asn His Asn Asp Thr His Gln Ile Leu
4785 4790 4795 4800
His Ala Leu Ala His Ala His Thr Thr Gly His Pro Ile Asn Trp His
4805 4810 4815
Thr Thr His Gln His His Thr Pro Thr Pro Gln His Ile Asp Leu Pro
4820 4825 4830
Thr Tyr Pro Phe Gln His His His Tyr Trp Leu Asn Thr Pro Thr Gln
4835 4840 4845
Thr Gly Asp Ala Ala Ala Val Gly Leu Asp Pro Ala His His Pro Leu
4850 4855 4860
Leu Gly Ala Ala Val Ala Val Ala Glu Gly Glu Gly Tyr Leu Leu Thr
4865 4870 4875 4880
Gly Arg Leu Ala Leu Ser Thr His Pro Trp Leu Ala Asp His Thr Ile
4885 4890 4895
Ala Gly Ala Val Val Leu Pro Gly Thr Ala Leu Leu Glu Ile Ala Leu
4900 4905 4910
Gln Ala Gly His Arg Val Asp Cys Trp Arg Ile Glu Glu Leu Thr Leu
4915 4920 4925
Gln Ser Pro Leu Phe Ile Pro Glu Glu Gly Ala Val Gln Val Gln Ala
4930 4935 4940
Trp Val Ala Ala Pro Asp Glu Asn Gly Cys Arg Ser Leu Thr Val Ser
4945 4950 4955 4960
Ser Arg Arg Glu Gly Thr Tyr Glu Asp Ala Thr Trp Val Arg His Ala
4965 4970 4975
Thr Gly Arg Val Gly Pro Ala Pro Ala Asp Gln Asp Glu Ala Ile Ala
4980 4985 4990
Arg Leu Thr Asp Pro Gln Gly Asp Gly Ala Ala Ala Ala Val Trp Pro
4995 5000 5005
Pro Gln Gly Ala Val Ala Phe Thr Ala Asp Asp Leu Glu Gly Leu Tyr
5010 5015 5020
Asp Gly Tyr Ala Ala Arg Gly Phe Glu Tyr Gly Pro Val Phe Arg Gly
5025 5030 5035 5040
Leu Arg Ala Ala Trp Arg Arg Gly Glu Asp Ile Phe Ala Glu Val Arg
5045 5050 5055
Leu Pro Asp Thr Ala Asp Gly Asp Ala Ser Gln Phe Ser Val His Pro
5060 5065 5070
Ala Leu Leu Asp Ala Ala Leu His Ala Ala Ala Phe Arg Pro Ala Asp
5075 5080 5085
Lys Leu Pro His Gly Ala Leu Pro Phe Ser Phe Ser Gly Val Arg Leu
5090 5095 5100
His Gly Pro Gly Ala Ser Thr Leu Arg Val Arg Leu Thr Pro Asp Gly
5105 5110 5115 5120
Gln Ala Arg Asp Thr His Ala Trp Ser Val Ala Val Val Asp Gly Glu
5125 5130 5135
Gly Arg Pro Val Ala Ser Ile Ala Ser Leu Ala Val Arg Pro Val Ser
5140 5145 5150
Thr Gln Glu Leu Leu Ala Ala Ser Gly Thr Ala Arg Arg Asp Ser Leu
5155 5160 5165
Phe Ala Val Glu Trp Val Thr Ala Leu Ala Pro Thr Ser Ser Ser Val
5170 5175 5180
Pro Gln Arg Leu Ala Thr Val Gly Pro Ser Asp Arg Leu Pro Ser Ala
5185 5190 5195 5200
Asp Ala Tyr Ala Asn Leu Ala Asp Leu Ala Ala Ala Val Leu Glu Ala
5205 5210 5215
Gly Ala Pro Ala Pro Asp Ala Val Val Val Asp Cys Gly Arg Arg Asp
5220 5225 5230
Ala Arg Ala Thr Ala Val Pro Glu Asp Val Arg Thr Leu Thr Arg Arg
5235 5240 5245
Ile Leu Gly Leu Leu Gln Glu Trp Leu Ala Asp Glu Arg Pro Ala Ser
5250 5255 5260
Ser Arg Met Val Val Leu Thr Arg Gly Ala Val Ala Thr Thr Pro Gly
5265 5270 5275 5280
Glu Asp Val Ala Asp Leu Ala Gly Ala Ala Val Cys Gly Met Val Arg
5285 5290 5295
Ser Ala Gln Ser Glu His Pro Gly Arg Phe Val Leu Leu Asp Leu Asp
5300 5305 5310
Pro Asp Pro Asp Leu Asp Gly Gly Glu Val Pro Pro Thr Val Val Pro
5315 5320 5325
Ala Ala Leu Ala Cys Gly Glu Pro Gln Ile Ala Val Arg Ala Asn Arg
5330 5335 5340
His Leu Val Pro Arg Leu Thr Arg Val Pro Ala Ser Val Pro Val Pro
5345 5350 5355 5360
Gly Arg Val Pro Val Pro Ala Ala Glu Ala Ala Asp Pro Asp Thr Thr
5365 5370 5375
Pro Thr Ala Phe Asp Pro Asp Gly Thr Val Val Ile Thr Gly Gly Thr
5380 5385 5390
Gly Thr Leu Gly Ala Met Leu Ala Arg His Leu Val Ser Arg His Gly
5395 5400 5405
Val Arg His Leu Leu Leu Ala Ser Arg Arg Gly Pro Asp Ala Pro Gly
5410 5415 5420
Ala Thr Glu Leu Arg Ala Glu Leu Ala Glu Leu Gly Ala Glu Val Thr
5425 5430 5435 5440
Val Arg Ala Cys Asp Thr Gly Asp Arg Gly Ala Leu Ala Asp Leu Ile
5445 5450 5455
Ala Gly Ile Pro Thr Gly His Pro Leu Thr Gly Val Val His Ala Ala
5460 5465 5470
Gly Val Leu Asp Asp Ala Thr Val Ala Ser Leu Thr Pro Arg His Leu
5475 5480 5485
Asp Thr Ala Leu Thr Pro Lys Ala Asp Ala Ala Phe His Leu His Glu
5490 5495 5500
Leu Thr Arg His Ala Arg Pro Arg Ala Phe Val Leu Phe Ser Ser Ala
5505 5510 5515 5520
Ala Gly Val Leu Gly Ala Ala Gly Gln Gly Asn Tyr Ala Ala Ala Asn
5525 5530 5535
Ala Phe Leu Asp Ala Leu Ala Glu His Arg Arg Ala Gln Gly Leu Pro
5540 5545 5550
Ala Leu Ser Leu Ala Trp Gly Leu Trp Glu Gln Gly Ser Gly Met Thr
5555 5560 5565
Gly His Leu Asp Arg Thr Asp Arg Ala Arg Ile Asn Arg Ser Gly Leu
5570 5575 5580
Ala Pro Leu Ala Thr Glu Asp Ala Leu Ala Leu Phe Asp Ala Ala Leu
5585 5590 5595 5600
Ala Gly Asp Arg Pro Phe Leu Val Pro Ala Arg Leu Asp Leu Arg Gly
5605 5610 5615
Ser Ser Ala Ala Glu Thr Pro Ala Pro Leu Phe Ser Arg Ile Ala Pro
5620 5625 5630
Ala Arg Thr Thr Arg Gly Arg Ser Pro Gly Ala Glu Gly Ala Ala Asp
5635 5640 5645
Leu Arg Thr Arg Leu Ala Ala Gln Asp Ala Ala Glu Gln Arg Asp Thr
5650 5655 5660
Leu Leu Thr Ile Val Arg Thr His Thr Ala Ala Val Leu Gly His Asp
5665 5670 5675 5680
Thr Ala Ala Ala Val Arg Pro Asp Gly Ala Phe Arg Glu Leu Gly Phe
5685 5690 5695
Asp Ser Leu Ala Ala Val Glu Leu Arg Asn Arg Leu Gln Thr Thr Thr
5700 5705 5710
Ala Leu Thr Leu Pro Ala Thr Thr Val Phe Asp His Pro Thr Pro Ala
5715 5720 5725
Ala Leu Ala Asp His Leu Arg Thr Gln Leu Cys Gln Asp Ala Gln Ser
5730 5735 5740
Ser Ala Ala Ala Thr Ala Met Ala Ala Met Ala Glu Leu Ala Arg Leu
5745 5750 5755 5760
Glu Ser Ala Val Ser Asp Ser Val Ala Leu Asp Asp Asp Thr Arg Ser
5765 5770 5775
Gly Leu Ala Glu Arg Leu Arg Ser Leu Ala Arg Lys Met Ser Ser Gly
5780 5785 5790
Arg Val Val Asp His Asp Gly Gly Gly Ala Ala Asp Leu Asp Leu Gln
5795 5800 5805
Ser Val Thr Asp Asp Glu Met Phe Glu Leu Ile Asp Lys Glu Val Ser
5810 5815 5820
Arg Asp
5825
<210> 79
<211> 15800
<212> DNA
<213> Artificial Sequence
<220>
<223> insert region of pCR2.1-Adu-milA1-Ac2 (FIG. 6)
<400> 79
tctagagatg acggaaatcc cctggctgag ccggcatctg ccggccgagt gagcggcgcg 60
tcgacagcgt tgccgcatca ctcccccggg cgccgggcgg tcagcacgac atagccgatg 120
tccggcgtcc aggggagcgt cgtgcactgt gcaaccacct tccgcatctc cgcgacgccg 180
atcccgaagg ccgccgcgta ctcgtcgagg tgctcgttca cgtggtcacg cagcgcggcg 240
aaagaggggc cgacgacctg atcgccgatg tcggtcagct catgcagttc cagcccggcg 300
tcggcgatca ttccggcgta ctcgtcgatg tgcaccaggg ccgggaccgc gagcaactgg 360
gacgtgcact ccccgcgctt catgccggtc cgcccgaagg cgcgcagcgt cacgtccgtg 420
acggccagcc ggccgccggg gcggagcacc cgggcgatct cccggatcac ctgtgcgggg 480
ctgggcatgt gcaggagaca ctccagggcc cacgccgcgt cgaaggaccc gtcgggaaac 540
ggcagttcca tcgcgtcggc acgggtgaac acgacccggt ccgccacgtg cgactgcttc 600
gcgagagcgg tcgccagccc gacctgaacc tcgctcaccg tcacgccgac gacatcgacg 660
ggcgcgctca gggcgagccg caccgccggc tttccggaac cgcagccgac gtccaggacc 720
cggcggcccg tgatgcctct cagcttgccg atgaggagat cggtgagccg gtcggcggcc 780
ttgcccggtg aactgccgtc ccccggctgc ggccagtatc cgaggtgggt gttcccaccc 840
agcgcacgat tcatgaggtc ggtcaaacgg tcgtagtagt cccccacttc cagggaagag 900
ggcggggtct gctccgggac ggccatcatg gtcgggaacc tccgcaatcc gggccgggcg 960
gcccagctgt cgtggcgatc tactccagga aacgtagacc tttttctgcc acttgtccga 1020
gctatgcaga caccccgatc ccctaagaaa tgaacaccct tgggaacggc acagcccagg 1080
ggtggatagg ggtattcgcc gccgccgcgc cgtcattagc tttgaagagt tgaagacgtt 1140
caagacattg atgcccggcc gtcagcggat ttctcgcgct cctttcattc ttcgacgctg 1200
cattgcagct ctcatcatgt ccgcacggcc gccgagcatt gcctagcggt gaggacacag 1260
ctcagttgcc caaagcccag aacgagttcg cagtggccgg tcatccgtgg atcctctccg 1320
ggcacaccgg aaccgcgctg cgggcccagg cacgccggct ccacgaccat gtcgccgacc 1380
accctcggct ccgtccggaa gacatcgccc acacgctggc gagcagcggc ccggcgctca 1440
cccatcgcgc ggcggtgatc gcggcggacc gggaaggaca tctccggggg ctcgacgcgg 1500
tggcccgggg tgaggacacc cccggtgtcg tacggggcac ggcggccgcg ggcggcgacg 1560
gggtcgcgtt cgtcttcccc ggccagggca cccagtggcc cggtatggcc gccgatctgc 1620
tgacggtctc ccccgccttc agccgggcgg tcgacgcctg cgccgaggcg ttcgaaccgt 1680
atgtctcctg gtcaccggag gccgtgctgc ggggcgctcc gggcgcgccg cccctggagg 1740
ggaccgatgt ggtgcagccg acgctgttcg ccgtcatggt ggggctggcc gagctgtggc 1800
ggactcttgg ggtgagcccg acgtcgatcg tgggccactg catcggggag atcgcggcag 1860
cccatctctg cggcgccctg tcgctgtccg acgcggcgcg cgtggtgatc gagagcagcc 1920
gggcccaggc gacgctctcc gggtcgggtg cgctgatcgc ggtcgcgcgg tccgaggcgc 1980
agctgcttcc gttgctgcgg cggtggccgg gcaggctgac gatcgccgcg gtcaacggcc 2040
cgatggccac ggtcgtctcc ggcgatcggc cggccgccga cgagctgttg gcggagttcg 2100
cccgtgccgg tgtccgggcc cgcgaggtgg cgatcgacat ccccgcgcac tcgccgttca 2160
tggcccccct cagggacggt ctgctcgact cgctgtcatc ggtcaccgcg ggtgcgtcgc 2220
ggctgccgtt ccactcctcg gtcatcgggg ggccgctgga gacccaaggg ctcgacgcgg 2280
cttactggta ccggaacctc gccgacacgg tccgcttcga aagcgtcgtc acggggctgc 2340
tgcggcaggg cacacgctgc ttcgtggagc tgagcccgca cccgatgctg accatgtgtg 2400
tgcaggccac cgccgaggag gtggtcggcg gtgagcgcgt cgtgatcctg ccgacgctgc 2460
atcgcgggca ggccgccgtc gagtccgttc gcaccacgct ggccgagctg tacgtacggg 2520
gcgcactgga tgaccatcgg gcggcgttct cggtgccggg cggccgcctg atcaccctgc 2580
ctctcgagcc gcccgcggac acgtccgtag agctcgccga cgccccggac ccggcggagg 2640
cctgccggcc ccccttggtg gagcggcttg cccggctctc caccgcggag cggaagcggc 2700
ggctgcgcga gctggtgggc gtcgaggcgg ccaaggtcct cgaggacgtc gccggggcgg 2760
acgcgccggg ccacggcatc gcggagcagg agcacttcgt cacttcgggc ttcgactccg 2820
cggccgcggt cgcgctgcgc aaccgcctga acgacgccac cggtttgctg ctgcccttca 2880
ccctggcctt cgaccatccg acacccgccg ccgtcgccga ccatctgcac tcccggctct 2940
tcgatcacca gggcggcggg cagccgggcg ccgacggccg gcccgacccc gcggcggcgg 3000
ccggtccggc cagggccgac gacgagccga tcgccgtcat cggcatggcg ggccgcttcc 3060
ccgggggcgc ccgtaccccg gaggagctgt gggaactggt cgccgaaggc accgacgccc 3120
tctcgccctt cccggagggc cggggctggg atccgctgcg gctctacgat ccggaccccg 3180
cccggcccgg cacgtactac cagcgcgaag cgggattcct ccacgacgcc gacaagttcg 3240
acgccgagtt cttcggcatc gcgccacgcg aggccaccgc aatggatccc cagcagcggc 3300
tgctcctgga gacctcctgg gaggcgctcg aacgggcgcg gatcgacccg accgcgctgc 3360
gcggcagccg caccggggtg ttcgtcggcg tggccccgct ggactacagc ccccgaatgc 3420
accaggcgtc gccggagctg gagggccatc tgctgaccgg caacatcggc gccgcggcct 3480
cggggcggat ctcctacgta ctcgggcttg aggggcccgc ggtgtccgtg gacacggcgt 3540
gctcgtcgtc cctggtcgcc ctgcatctgg cggcccaggc gctgcgggcc ggggagtgct 3600
cgctggccct ggtcggcggg gcgacggtcc tctcgacccc cggcatgttc atcgagttct 3660
cgcggcagcg cggtctggct ccggacggcc gctgcaaggc gtacgcggcc gccgcggacg 3720
gcaccggctg gtccgagggt gtgggcatgc tgctcgtcga gcggctgtcc gacgcgcgac 3780
ggctcggaca ccaggtgctt gcggtggtac ggggctccgc cgtcaaccag gacggggcga 3840
gcaacggctt cacggcgccc agcggtccat cacagcaaca ggtcatccgg gcggccctgg 3900
ccaatgccgg ggtgtcggct ccggaggtcg acgcggtgga ggggcacggc accggcaccc 3960
ggttgggcga tccgatcgag gcgcaggcgc tgctggccgc ctacgggcag gggcgggcgg 4020
ccgaccggcc gctgtggctg gggtcgatca agtcgaacat cggacacacc cagtgggccg 4080
cgggtgtcat cggggtcatc aaaatggtgc tcgcgctcca gcacggtgtg ctgccgcgca 4140
cgctgcacgt ggacaagccg tcggattacg tggactggtc ggccggggcc gtacggctgt 4200
tgacggagcc ggtgccctgg ccggagcggg gccacccgcg ccgggcgggg gtgtcgtcct 4260
tcggggtgag cggcaccaac gcccatgtca tcctcgagca ggcaacgcca tcgtccacgg 4320
tggctcccgg ggggcatacc gccgaggccg ggcctcccct gccgtgggtg gtctcggcga 4380
agacgcccca ggcactgcgc gaccaggccc gccgcctgca cgaacacctc accgcccagc 4440
cacagctcca accggccgac gtcggccaca ccctcgccac cggccgcgcc accttcgacc 4500
accgggccgt cctcatcggc tccgaccgcg aacaactcct ccacggcctg gacgcgctcg 4560
ccaccggccg gcccgaccca gcggtccacc agacgtcgga ccgtcccgcc accgccgacg 4620
gccgtatcgt cttcgtcttc cccggacaag gcggtcaatg ggcgggcatg ggcctacggc 4680
tgctgaacgc ctcacccgtc ttcaccgagc ggatggccgc ctgcgaacag gccctctccc 4740
cctacgtcga ctggtcactc acggacatcc tccaccggcc ggccgacgac gccgtatggc 4800
aacgcgccga catcgtccag cccgccctgt tctcgatcat ggtgtccctg gccgcgctct 4860
ggcgctcttg cggcatcgaa ccggacgccg tcctcggcca ctcccaaggc gagatcgccg 4920
cggcccacgt ctgcggcgcc ctgacgctcc acgacgcggc caaggtcatc gccctgcgca 4980
gccaggccct ccaagccgta cgcggcgccg ggggcatggc ctccgtaccc ctgcccgcgg 5040
accaggtcac cgaggatctg cgcacccact ggcccgaccg gctatgggtg gccgccacca 5100
actcccccac ggcaaccgtc atctcgggaa acaccgacgc gcttgacgaa gcgctcgacc 5160
actaccacgc ccacgacgta cgggccaagc gcatcccggt cgactacgcc tcccactgcc 5220
cccatatcga cgcggtggcc gagcgactgc ccgacctgct gggcggcatc gtcccgcgcg 5280
ccgccgacat ccccttctac tccacggttg acggccgatg ggccgagccg accgagctcg 5340
acgccgacta ctggtaccgc aacctccgca gcccggtacg gttcgcccac gccgtccacg 5400
ccctcaccga gaccgaccac cgcacctttg tcgaagtcag cccacacccc acgctcaccc 5460
ccgccatcac ggccaccacc gaaaccaccg accgcaccac caccgtcatc gcctcgctcc 5520
accgcgacca cgacgacacc caccacatcc tcaccaacct cgcccaggcc cacatccacg 5580
gccacaccat cgactggcga caccactacc agactctgcg ccccacccca ccccatatcg 5640
acctccccac ctaccccttc caacaccacc actactggct ccacgactcc accgaggaca 5700
aggcggtggg tacggacctc gccgcggccc gcttctggga ggcggtccac ggcgaggaca 5760
ccaacgccgt cgccgcgctc ctcgacgtcg agccgggcac ctcactggac gcgctgctgc 5820
cggccctgtc cgcctggcac ggtcggcgtc gcgaccaggc catcaccgac acctggtgtt 5880
accgcgacat ctggaagccg gccgacctca ccgccgcgcg cccccggccg tccggccgat 5940
ggcttgtcgc gatctccgca gggcgggccg atcacctcca cgtcagtgcc gtcctggacg 6000
ctctggaacg ccagggtctg cccatcgcca ccctcgtcct cgacgacacc cacaccgaac 6060
tccccctgct ggagcggcat ctcgcacagg cgatcgcgag cgatgggccg gccatcggcg 6120
gcgtgctctc gctgctcgcc ctcgacgagg ggccacatcc gcgccacccg gaggtgcccg 6180
tcggcaccgc cctcaccctc agcctgatcc aggcgctcat cgcacgcgag gacatggcgc 6240
cccggctgtg gctggccacc cacgaggccg tcgccacctc gtccgcggat acgctcgatc 6300
accccctcca ggcgatggtc tgggggctgg gacgcaccgc cgcactcgaa caccccgatc 6360
tgtggggcgg cctcatcgac cttccggaca ctctcaccga acgggtcctc cacggcctcg 6420
tcacggcgct gaccacctgt cacgacgagg acgaactcgc gctgcgcgcc accggcccac 6480
gcacccggcg cctgatccgg acgccgtcca ccgccgcagc ggaggacacc ccgccgtgga 6540
cgccccgtgg caccgtcctc atcaccggcg gcaccggggc cctgggctcc cgcgtcgccc 6600
accgcatcgc cgaacgccac cccgactgcc acttgctgct ggtgagccgg cgagggccca 6660
aggcccccgg cgccaccgcg ctccgcgacc agctcatcga actcggcgcc acggtgaccc 6720
tcgccgcatg tgacaccgcc gaccccggcg cgctcgcgga tctcctcgcc gatgtcccct 6780
cggaccgccc cctcaccgcg gtcgtccaca ccgcgggcgt cctggacgac agcaccctcg 6840
ccgtacagac cccggaccac ctcgccgccg ttctggggcc caagtcccat gccgcacacc 6900
atctgcacgc cctcgcccag caccaccccc tcgacgcgtt cgtcctcttc tcgtccgtcg 6960
cggcgccctt cggcgccgcg ggccaggcca actacgcggc cgccaacgcc tacctcgacg 7020
ccctcgccca gcaccgccgg gcccaggggc tggccgccac ctccatcgcc tggggcaact 7080
gggacggcga cgggctcgcg agcacacagt ccgcgcagac gtacctgcgc aaccgcggct 7140
ttcctccgat gccgccacac ctggcgctgg ccgccctgga gcgagccatc gtctcgcccc 7200
acgcccagct cgtcgtcgcc gacgtcgact ggaagaagct caagccggcg ccgcacaccc 7260
gcgacatccc gggaagccgc cgcccggccc cggccgccac cgacggcgcg gacaggacgg 7320
ccgacgccac cgcgagcctc cgtacccgcc tcgcgggtca gagcccggcc gaacggcacc 7380
agacgctcct cgacctcatc agctctcata cagccgccgt cctcgggcac gccacgcccc 7440
agacgatccc cacggaccgg gccttccgcg acctgggttt cacctcgctg acggccatcg 7500
agctccgcaa ccgcctcgcg gcggccaccg ggctccgcct gccgaccacc gtcgccttcg 7560
accgtccgac gccggacaag ctcgcggccg acctgctggc gcggtgcgcg ccgacaggcc 7620
cggacggcat cggggtgacg cccgacgcga cggccacgag tggcagttcg cccggtgcgg 7680
cgcatggcgc gccggacccc gccgagccca tcgccatcgt cggcttggcc tgccgctacc 7740
ccggcgggat cggctccccc gaggacctgt gggagttcat caccgcacac cgggacgccg 7800
tcggagactt cccgaccgac cggggctggg acctggcgag gctcttcgac cccgatccgg 7860
accggccggg cacctcgtac agccgacagg gcgccttcct ccgcgacgcg ggcgacttcg 7920
acccggagtt cttcgggatc agcccacggg aggcgacggc gacggacccc cagcagcgac 7980
tgctcctgga ggcgtcctgg gaagccctcg aacgagccgg gatcaacccc cacgatctcc 8040
acggcagtcc gacgggcgtc ttcaccggca gcaacgcgca ggacttcagc gcgcggctgc 8100
ggcagacgcc gtcggagctg gcggagctgt gcgagggcta tgcgctgact ggcagcaaca 8160
acagcgtcgc ctcggggcgc gtctcgtacg cgctcggcct ggaaggcccg gcggtcagca 8220
tcgacaccgc ctgctcgtcc tcgctcgtgg cgctccatct ggcctgccag tcgctccggg 8280
ccggcgaatg ctcgctggcc ctggcgggcg gcgtcacggt catgatgacc ccgttcaact 8340
tcgtggagtt ctcccggcag cggggcctgg cggcggacgg ccggtgcaag gcgttctccg 8400
ccacagccga tggcaccggc tggggcgagg gcgtgggcat ggtggtggtg gagcggctgt 8460
cggacgcgcg gcgcaacggc catcgtgtgc tggccctcgt ccgcggcagc gccgtcaacc 8520
aggacggtgc cagcaatggg ctgactgccc cgaacggccc ctcgcagcag cgggtcatcc 8580
gcgccgccct ggccgccgcc ggggtcgccg cggcagaagt ggacgcggtc gaggcgcacg 8640
gcacggggac gacgctcggc gatccgatcg aggcccaggc cctgctcgcc acctacgggc 8700
aggggcggcc ggcggaccgg gcgctgtggc tcggtacggt caagtccaac atcggacacg 8760
cccagtcggc cgccggtatc gccggggtca tcaagatggt gctggccctg cggcacggga 8820
tgctgccgcg tacgctgcat gtgtccgagc cgtcgccgca tgtggactgg tcggcgggtg 8880
cggtacggct gctgaccgag gaccagccgt ggccggacac cgggcgcccc cggcgggcgg 8940
gggtgtcgtc cttcggcgtg agcggcacca atgcccatgt gatcctggag caggcggagc 9000
cggggccgga cccggcaccg acggcctccg cgccctccct gcccccctgg cccctctccg 9060
ccaggtcggc ggaggccctg cgggcccagg cccgtaggtt gctggcgtac gtggccgagc 9120
acccggatgt cgaccccgcc gacgtggggt actccctcgc gcgcggacgg gccgtgttcg 9180
agcaccgggc cgtgctcctc ggcaccggcc acgacgactt ccggcgcgcc ctggacgccc 9240
tggcgtcggg cgcgcccgac ggcgcggtcg tccagggcgc ggcggtgggg cggcagggca 9300
aggtcgtctt tgtgtgctcg gggcagggca cccagcgccc cggcatgggc cgcgggctct 9360
accgctcgtc cacggcgttc gccggggcgc tggaggaggt gtgcgcccat ctggacccgt 9420
atctggaaca ccctctgatg gaggtgatgt tcgccgatga gaagagcgat acgtcggcgc 9480
tgctgcatct gaccgcctac gcccaaccgg ccctcttcgc cctccagacc gcgctgcacc 9540
gcatggtcac cgaggagttc gggctcaccc ccgactatct ggccggccac tccctgggcg 9600
agctgaccgc cgcccatctg gcgggcatcc tcagcctgcc cgacgccgcg gcgctggtgg 9660
cggcccgcgc ccgcgccatg cgggaccttc cagcgaccgg agccatggtc gccgtcgagg 9720
ccaccgaggc ggagctgcgg ccccggctcg ccgagttggc ggaccgggtc ggcatcgccg 9780
ccgtcaacgc ccccgcgtcc ctggtcatca ccggcgacca cgacgccgtg caccagatcg 9840
ccgacgactt ccgcgggcag ggcaggaagg tcactcccct ccaggtcagc ggcgccttcc 9900
actctcccca tatggagccc ctgctcgacg agatcgggcg caccgccgaa accctcacct 9960
accaccggcc ccacactccc ctcgtcaccg cgtcggccga cggcggcgac gacacgaccg 10020
agccgcgggc cgacgacgac ccgggcacgg ccgcgttctg gcctctccag gcccggcgca 10080
ccgtccacta cgcgcgggcc gtggagcggc tgcgcgcccg cggcgtcacc acgttcctgg 10140
aactcggccc cgactccacc ctcactaccc tcgtccacca caatctcgcc gcgcacgatc 10200
ccgtggccgt ctccctgctc catccggagc ggtgcgagac gcacagcgtc ctcggcgcac 10260
tcgccgcggt ccacgcccac agccgccccg tcgactggac acgccactac accgcacggc 10320
cgcggccgac gccacaccag atcgacgtgc ccacctatgc cttccggcac cggcgctact 10380
ggctgcccgc cccggcggcg gtcggcgatg tgacggccgc ggggctcgac gcggcggagc 10440
acccgctgat cggcgccgcc gtgtggctcg cggagggcga cggctgtctg ctgaccggca 10500
ggatctcgcc gcgtacgcac ccgtggctgg ccgaccatgt catcgccggc actgtgctcc 10560
ttccgggcac cgcgttcgtg gagctggcgc tgcgggccgg ggcgtacgtg ggctgcgacc 10620
gtgtggagga gctgaccctg cacgcgccac tcccgctgcc cgccgacggt gaggtggtgc 10680
tgcaggtggc ggtgggggcc gccgacgagt ccggccgccg tgagctgagc atccacgccc 10740
ggccggcgga cgacggtaca tggacacggc acgccatcgg cacgctggca tcggcccgcg 10800
gcgtcggcct cgacgatggc acggggcaca atggccacgc cccggcgggc gacgagccgt 10860
tcgggtcgtg ggccacggcc tggccgccgc ccggtgccga gcccttggac gtcaccgggg 10920
tctacgaccg gtttgccgac gccgagttca cgtacgggga ggcattccag gggctggtcg 10980
cggcttggcg gcacggcgac gagacgctgg cggaggtccg cctccccgac cagccggccg 11040
gtgacgccct ccgcttcggg ctgcaccccg cgctgctcga cgcggcactg cagaccatgt 11100
ggctcgtgga gcccgacggc acacggccga gcggtggcct gggcggcccc gatcggggcc 11160
tgccgttcgc ctggcagggg gtctcgctgc gtacggcggg cccgtcggcc ctgcgggtac 11220
ggctgcgacg gccggcgccg gacaccgtgg ccgtcgccgt ggccgacgcg gccggccggc 11280
cggtcgcgtc ggtggagtcg ctgacgctgc ggccggtgcc gcggggcgcc ttgcgcggca 11340
ccgagacggc ggtgcgcacc tcgttgtacg gcctggactg gacggatgtg ccgctgccga 11400
cgccgcagac ggccctgccc cggtgtgcgc tgatcggagc ggacacgctc gacctggtcc 11460
ccgcgctcga ggccgcggcg cccgaccgca tcaccgacgg cgtggagcgc tacgccgacc 11520
tggaggagct ggtgcgctcc gtggcggcgg gcgcccccgc cccggacctc gtcatcgccg 11580
gctgccacgc agcccctgaa gccgacggcg cgagcgaaca gccacagccc gagacggtgc 11640
gcacaaggac gggtcaggtg ctggagctgc ttcagcggtg gctcggcgcg gacgggctcg 11700
ccgacgcaca cctggtgctg ttcacctcag gcgcggtcgc cacccggccg ggcgagccgg 11760
tgcgggacct ggcgggggcg gcggtctggg gtctggtgcg ctccggccag tcggagcatc 11820
cggagtgctt caccgtggtg gacatggacg gcgcccagga gtcccgcgcg gcgctgctcg 11880
gcgcgctcgg cctcggcgag ccgcaactgg cggtgcgcgg cggccgggcg ctggcgccgc 11940
gcctggtgcg cccgggtgac gccgacgacg acagcggcct ggccctgccg caggggccgg 12000
aaggctggcg gttggagtgt cccggcacgg gcagcctgga cgggttgacc acgaccgagt 12060
ccccggccgc ggcggtgccg ctcggcccgg gcgaggtacg ggtcgcggtg cgggccgcgg 12120
ggctgaactt ccgcgatgtg ctgatcgcgc tgggcgtggt gcccgggcgg acggcgctgg 12180
gcagtgaggg ggcggggatc gtcctcgagg tcggggcgga ggtccgcgat ctcgcgcccg 12240
gggaccgggt ggtgggtatc ttccccgagg cgttcggccc ggtggccgtg gccgagcggg 12300
cgaccctggc gcgggtcccc gacggctggt cgttcgccca ggccgcgtcg gtccccatcg 12360
tgttcgccac cgcgtaccac ggcctggtcg atctggcgcg cctgcggccg ggggaatcgg 12420
tgctgatcca tgccgcggcc ggcggggtgg gcatggccgc cgtgcaactg gcgcgccatc 12480
tgggggccga ggtgtacgcc acggccggcc ccggcaagtg gcacatcctg cgttcccaag 12540
gcatcgacga cgaccatctg gcctcgtcgc gcacgctgga gttcgagcag cgcttcgccg 12600
cgacccgcgg cgggcggggg atcgatgtcg tcctggactg tctggcccat gagttcgtcg 12660
acgcctcgct gcgcctggtg gcgcgtgacg gcggccggtt cctggagatg ggcaagagcg 12720
acatccgtga cccgcggcag gtggcgctgg accatccggg cgtgctctac cgggcgttcg 12780
acctgctgga ggccgggccg gagcgggtcg ggcagatcct gcgcaccgta ctggacctgt 12840
tcgagcgcgg tgtcctggcg cacctgccga cgacctgctg ggacatccgg caggcggagc 12900
acgccttccg ccatctgcag cagggccgtc acatcggaaa gaacgtgctc accgtcccgg 12960
ccggctggaa cgccgagggc accgtactga tcaccggcgg tatgggcacc ctgggcgccg 13020
ccctcgcccg tcatctggcg ggtaccgggc gcgcccgcca tctgctgctg gccggccgac 13080
gcggccccga cgccccgggc gccgaggagc tgcgagagga gctgaccgag ctgggcgcgc 13140
gggtcaccat cgccgcatgc gatctcggcg accgggcggc ggtcgcccgg ctcctggggg 13200
cgatcccggc cgagcggccg ctgaccgctg tcatccacgc ggcgggtgtc gtcgacgatg 13260
ccaccctcgg gtccctcacc ccccgccacc tggacgccgc cctggccgcc aaggccgacg 13320
ccgcctggca tctgcacacc ctcacccgcc acgccgacgt ggccgcgttc gtcctcttct 13380
cctcggtcgc gggtctgctc ggctcgcccg ggcagggcaa ctacgccgcg gccaacgcct 13440
tcttggacgc gctcgcccac caccggcgcg gctctggcct tccggcggtg tcgctggcgt 13500
gggggctgtg ggagcagacc agcggcatga ccgggcacct ggaccaggcc gaccgcgccc 13560
ggctggcccg gctcggcatc agcccgctca cgaccgggca ggcgctcggc cttttcgacg 13620
ccgccctcgg ccaccaccgc cccgtgctcg tccccgcccg cctcgacgtg cccgatccgc 13680
accccggctc gtcgaccgtg ccgcccctgt accggggcct ggtcggatcc aggacccggc 13740
ggacaccccc cgcggccgcc gccaccgggc cgttccccct gcatacccgc ctcggcggtc 13800
acgccccggc cgagcagcac gagatgctgc tctcgctggt ccgctcccac gccgccctcg 13860
tgctgggccg cgacgatccg gacacggtcc atcccggcgc gcacttccgc ggcctgggct 13920
tcgactccct gaccgcggtc gagctccgca accggctcaa cgccgccacc ggcctccggc 13980
tctccaccac cctcgtcttc gaccacccca cgcccgacga actcgcccgt cacgtccggg 14040
agcaggtgct gggcgacggc gaagcggcgc gggtggcccc cgtgttggcg gagctcgaca 14100
aactcgaatc cgccctctcc gccctcgaca aaaccgacag cgccagcgaa agagtcaccc 14160
tgcggctgaa gtcactcatg ttgaggtgga acgcacccca gcatccgaca gccgaaagcg 14220
ctgatgacga cgagaagttc acatcggcaa cagaggctga gattttcaaa ttcattgaca 14280
acgacctcgg cctgtcctga accggacgcc tgccactccg cccgtatccg ctgggccctg 14340
ctaggacgtg aatgcaattg gcgaatgaag cgaagctcct ggaatacctc aagcgcgtca 14400
ctgcggacct ggaccgcact cgccgtcgcc tgtacgaggt ggtcgagcgt gagcaggagc 14460
cgatcgcgat tgtggggatg gcgtgtcgtt acccaggcgg ggcgacgtca cccacgcgac 14520
tgtggcatct cgtcaagtcc cagacggacg ctatcgggga gttcccgacc gaccgtggat 14580
ggaacctgga gcagctctac gacccggacc ccgaccgctc aggaaccagt tacacgcgca 14640
gcggagggtt tctctatgac gcgggcgact tcgacgccgc gttcttcgag ttgtcaccgc 14700
gtgaggcgct ggcaatggac ccgcagcagc gcctgctgct cgaaaccact tgggaaacgt 14760
tcgaacaggg cggaatcgac ccgaggtcca tgcgcggaag ccggaccggg gttttcgtgg 14820
ggatcaatcc ggaggactac accaccggat acacacatca gccctcaaac gcagtcgagg 14880
gctacctgct cactggcagc gcggcaagca ttgcgtcagg ccgtatctcc tacaacttcg 14940
ggctcgaagg ccctgcgatc actatcgaca ccgcgtgttc ctcctcgctc gtcgccctgc 15000
atctggcctg ccaagcgctc cggtccggtg aatgcaccat ggcgctcgca ggcggcgcct 15060
ccgtcatggc cactcccttc gtcttcaccg agttctctcg ccagcggggc ctggccgcag 15120
acggccggtg caaggcgttt tcggcggcgg cggacgggac cggctggtcc gagggtgtgg 15180
ggatgctgct ggtggagcgg ctctccgacg cccgccgcaa cggtcaccgt gtcctggccg 15240
tcgtccgcgg cagcgccgtc aaccaggacg gcgcaagcaa cggcctgacc gcacccaacg 15300
gtcgttcaca agtcaaggtc atccgccagg ctttggccaa cgcacacctc tcccctgccg 15360
atgtcgatgc ggtggaggcc cacggcacgg ggaccaccct gggcgacccg atcgaggctc 15420
aagccctcgt cgaagcctac ggtcaggacc gccccaacgg ccgccccctc tggctcggaa 15480
ccctcaagtc caacatcggg cactccatgg ccgctgcggg tgtgggcggg gtcatcaaga 15540
tggtgatggc gctgcggaat ggtctgctgc cgcggacgtt gcatgtggat gagccgtcgc 15600
cgcatgtgga ctggtccgcg ggtgcggtgc agctgctgac ggagacggtg ccctggcccg 15660
gcggggaggg gcggctacgg cgggcaggag tgtcatcatt cggcgtcagc ggcaccaacg 15720
cccacgtcat cctcgaggaa gcacccgccc acaacatccc gtcagacaca cccgccgacg 15780
acgtcccggg agaatcagcc 15800
<210> 80
<211> 22878
<212> DNA
<213> Artificial Sequence
<220>
<223> nucleotide sequence of pD7K9milA3
<400> 80
ctatgcaggt ccccggggat cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc 60
cgcgaagtcg ctcttcttga tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc 120
ccggccgttt tagcggctaa aaaagtcatg gctctgccct cgggcggacc acgcccatca 180
tgaccttgcc aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg 240
catcaccgaa ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc aggccgccca 300
ggcggcccag gtcgccattg atgcgggcca gctcgcggac gtgctcatag tccacgacgc 360
ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc cgacaggctc atgccggcca 420
gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa 480
ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat 540
acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga 600
tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagat 660
ccgtcgacct gcatactagc tgctcaaggt cgcccgcaac cggcgcatca agcccgccgt 720
agctggtctc gcccgcgccc tggccgacgc cgccgcccgc gccgccgccg acgacgaacc 780
gtggacctgg accatcagcg cctaggtttc tgcactttct gcactgccca acgcgggcgc 840
gcgtgtgcgc cggccaccgc ccggggtgta acggaaatcg ccagcagacg tagtccttgg 900
tgagactccg ccccaacgcc ctgatctgcg aagaggaccg ttcgtggaac tgatgagacc 960
gctctacgcg agcgcgcgcg gctgtaacgg atcacgaggt cactccgtcc ataggccgaa 1020
gaggctcacg acgaggaggt cctgacgcac aggaagcccc gcccaggccc gggcggggct 1080
tcttccgacc tatcaaatcc caccagcacc gggcgtatcg gacgtctatc ggcgcgtcag 1140
gagcgagcag gggcaatgaa cggccacacc aatgccagga tcaacaggac ccagaaatgg 1200
cacgagcccg ggattggcct cccgggctcg ccgcccccga accggcgcag catgccgagt 1260
gaggacgttt gctctgtgac gagcgtacga gagaacagcg cgtcgacacc accgcgcccc 1320
gccggaccga agtgccactg cggcaccccg atcgtccaca tccccgggaa gcgccccaaa 1380
atctactgct cggacgcctg ccgaaagcgc gcaaagcgtc tcattgcgga ccagtcacgc 1440
agcgtagcaa gcacgactac agcgggtaat ccggacgcgg gctttagtag taagggaaaa 1500
tctccgtcca ctgcaggtca gagcgctgta actggaacgg gttctacccc aaacgagggt 1560
gccgcgacag ccgggcggga ggacccgcgc gacgagcggt tcgcccggcg ggaccggcac 1620
cagacggtgt cgctgaacga ggcgttcaag ggctgcggga cccggctcac tgccgggacg 1680
gccgagctga tgtggaagcc gggcgaggcg acgtggggca acacgtgccg ctgcaacaac 1740
gtccacacct gcccgtggtg catgagccgc atcctggccg tacggggctc gaacgtgcag 1800
ctcgccgccg acggtctcgc ggacgccggg tacgggctgc accttggcac gaacacgttg 1860
cgccacttcg agcgcatggc gttcggcacc gtccgcaagg gaatgcggca cggcctggtc 1920
gcggtcttgc acgacggctg gaagggcgcg tacggctcgt ccggccgccg ctggcgcacg 1980
atgcgcgacg acttcgggat catcggctac gagcgggcct tcgaggacac cttcggctgg 2040
ggctccggct ggcacctgca ctggcacacg ctctgggtca cccgcgaggt gctcggcccg 2100
gacgcccagg ccgcgttccg cgacgcactc gccggagcct gggccgccgg ggtcgaatcc 2160
gcgggcggct acaccgtcag cgagacgtgc gaccggcccg gttgctcctg tgagggcaag 2220
ggccacggca ccgacgtgcg cccactcaac ggcgcggacg ccgctgacgg cgatgcgggc 2280
aagcaggccc gctacctcta caaggacggc gacaagacca agggcggcgt cgccaagatc 2340
ggcctggagc tggccggaca gaacttcaag gccggtcgcg gcgacgaccg catgggaccg 2400
ctcgacctgg gcgacgcggc ggccgccgag ctgcagcggc ttcgtcggcc cggcccgttc 2460
gtcgagaagt accgcgagcg cgagttcggc gtcttccagg tccgcaagca ctaccggtcg 2520
cagaacctga accggctgat caaggaactc ggcatccagc aggacgtccg caccgaggag 2580
gagatcaccg acgacaccga gggcctggtc gccatcgcgg tcatcccggc ctacatctgg 2640
taccgataca tcgcccgcgt cgccggacgt cgcctcgacc tcatcaaggt cgccgagacg 2700
tacggcctac cgggcgtccg gcggctcgtc gagtcctggg gcctggtgtg gggcaaggac 2760
gtcctggacc cgcccgcccc ggaagcgccg gccgcgccgg gcgaccttga cgctgaccag 2820
atgaggttcg aggtcatgtc cgaggaggaa gctgctttcc gcgaggcgcg ccggaaggcc 2880
aacgaggccc gcacggagga gctggccgct tcgctcgacc gagtgcgtca gccgaagaag 2940
gaggcgatcc gcccgacgat cagccttcgg aagcgcctga agccgaagcc ggtcaccgtc 3000
gacgtcaaga ctcccccgcc cggcgccgcg tcgcccgtgt gccgccgctg caagggcaag 3060
ctcgccccgg tactccagcc ctggggccgg ctccccggag actgcctccg cgtcgacaca 3120
gcggtcgcgt agggccgctg agggaccgcc atgagaccgc caccccggac atccgggtgg 3180
cggtctcatg gcggtcccgg cgcgaaagcc gagggtctgc ctgccgtgag gtggccggcg 3240
gacctgggtt actgctggaa tcccgcgccc agtccgagcg tccgccggcg cttttcacct 3300
ggctccgggt gctcgtcgat gagcaccagg ccgtcgacga cgacgcgttc ggcgggcagg 3360
agccgcgggg ccgacgtcga caacgacgtc ctggtggtgg tcgacggacc ggtgccgagt 3420
gcgcggcgag gggtgctcat gccgatgagt ctgaacttct ccggcgaagg gttctaccaa 3480
gtcctgggtt gtgttcgcct tggtagcgct cctagttggc gaggctggtc ctttaccctg 3540
atgccctcgg gcatcagggt aaaggaccag cctcctcaca tgagtaggct gaaacagcag 3600
ttcgttccgc ctcttcttcc aaagttgaga agccgcagaa tccctttctg gagggaaacc 3660
agcgtacgta cattgcactt tccctccagg cgagggtttt gcagatgctt ggcggtgtgc 3720
actgtctcta gctggcatgg ctttggaaat agctccgctc agctccgagg gcttctctgg 3780
ggtggctccc ctgaggaacc tcgacatcga tacgtcgcgg tgagttcagg ctttttcata 3840
tctcattgcc cccggacgag cgtctgctcc gccattcgcc gtccgccgtg ccaatcggat 3900
cagccgtcca aatgcgggat tttcgttagt cggaggccaa acggcattga gcgtcagcat 3960
atcatcagcg agctgaagaa agacaatccc cgatccgctc cacgtgttgc cccagcaatc 4020
agcgcgacct tgcccctcca acgtcatctc gttctccgct catgagctca gccaatcgac 4080
tggcgagcgg catcgcattc ttcgcatccc gcctctggcg gatgcaggaa gatcaacgga 4140
tctcggccca gttgacccag ggctgtcgcc acaatgtcgc gggagcggat caaccgagca 4200
aaggcatgac cgactggacc ttccttctga aggctcttct ccttgagcca cctgtccgcc 4260
aaggcaaagc gctcacagca gtggtcattc tcgagataat cgacgcgtac caacttgcca 4320
tcctgaagaa tggtgcagtg tctcggcacc ccatagggaa cctttgccat caactcggca 4380
agatgcagcg tcgtgttggc atcgtgtccc acgccgagga gaagtacctg cccatcgagt 4440
tcatggacac gggcgaccgg gcttgcaggc gagtgaggtg gcaggggcaa tggatcagag 4500
atgatctgct ctgcctgtgg ccccgctgcc gcaaaggcaa atggatgggc gctgcgcttt 4560
acatttggca ggcgccagaa tgtgtcagag acaactccaa ggtccggtgt aacgggcgac 4620
gtggcaggat cgaacggctc gtcgtccaga cctgaccacg agggcatgac gagcgtccct 4680
cccggaccca gcgcagcacg cagggcctcg atcagtccaa gtggcccatc ttcgaggggc 4740
cggacgctac ggaaggagct gtggaccagc agcacaccgc cgggggtaac cccaaggttg 4800
agaagctgac cgatgagctc ggcttttcgc cattcgtatt gcacgacatt gcactccacc 4860
gctgatgaca tcagtcgatc atagcacgat caacggcact gttgcaaata gtcggtggtg 4920
ataaacttat catccccttt tgctgatgga gctgcacatg aaccaaaagg atctaggtga 4980
agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 5040
cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 5100
tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 5160
agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 5220
ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 5280
acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 5340
ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 5400
gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 5460
gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 5520
gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 5580
tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 5640
caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 5700
tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 5760
gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg 5820
agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt 5880
ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc 5940
gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc 6000
ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct 6060
atgacatgat tacgaattca tacggctacg aggatggggc cgatgaagcg gctcgcgccc 6120
tgtgcgccct tgctccgggt gaaatcgccc gatccggagg agaccagcat ggtccgcacc 6180
aggtccgggt gcgtggcgag gtaggcggtt ttcggcccga ggcggagctt gagcaggtct 6240
ccgtgatctg ctgcggagcg aaggaactcc aggggctgcc gcatcaacgg cggcacatgg 6300
ccgacgaccg gccaggcgcc gggcgcctcg ggaatgctgc tcgtcgactg ggacatcacg 6360
agtgctcctt tgcggggtga aggggggtgg ctgggagggg aacgacagtg acgagtgaag 6420
ggggaggtgt gggggttggc gtcggcccgg gggtgagcgt ggacatggga gtgggaggga 6480
gtgaagtgag ctcggagtgg tttctgggct tcattgagat tcgaatccga cttccctgtc 6540
gatgagagcg aacatctcct cgtccgatgt ctctgcgagg tcgggggcgg tgtcgtcgcc 6600
gttcaacttc tgggcgaggg aatgcagtcg ggaggccagc cgcgtgcgcg cgccgtcgtc 6660
cagcggggca gcggaggatg tggtggagga gagcactacc gcctccagcc gctcgagctc 6720
cgagagcagc gagggcagcc ccgggggtgt cgttgcggcg tccggctcgg ccgcggcggt 6780
gagtcccttg ctgacgagtt gtgtgtggag gtggtgggtg agggtggtgg ggttggggtg 6840
gtcgaaggcg agggtggtgg ggaggcggag tccggtggtg tgggagagcc ggttgcgtag 6900
ttcgacggcg gtgagggagt cgaagccgag gtcgcggaac gcgcggtcgg gggggatggt 6960
gtcgggggtg gtgtggccca ggacggtggc gatgtgggag cggaccaggg cgaggagggt 7020
ggtgtgctgt tgttcgtgtg tctggccggc cagccggccg tgcagctggg cgccgttgtc 7080
cgcaccaccg gtagtggtgg tgcgggtggt gcggcggcgg gtggcgggca ggaggtcctg 7140
cagcaggggc ggcaggggcg gggcgggacg caggtcggcg ggcagcagga ccggccggtc 7200
cagagccagg gccgcatcga agagggccag tgcgtccggg gtcgacatgg gatgcaggcc 7260
tgagcggttg atgcgggccc ggtcggtgcg gtcgagatgc ccggtcatgc cgctgccctg 7320
ctcccacagg ccccacgcga gcgacaaggc cggcaggccc tgcgccctgc ggtgttcggc 7380
gagggcgtcg aggaaagcgt tggcggccgc atagttgccc tgcccggctg cgccgaggac 7440
accggcggcc gaggagaaca ggacgaaggc gcgcggccgg gcgtggcggg tgagctcgtg 7500
cagatggaag gcggcgtcgg ccttgggtgt cagcgcggtg tccaggtgtc ggggggtgag 7560
cgaggcgacg gtggcgtcgt ccaggacgcc cgcagcgtgg accacaccgg tcaaagggtg 7620
gccggtggga atccccgcga tgagatccgc cagcgcgcct cggtcaccgg tgtcacaagc 7680
gcgcaccgtc acctcggcgc cgagctcggc cagttccgcc cgcagctcgg tggcgccggg 7740
tgcgtcgggt ccgcgtcgcg atgccagcag gaggtgtcgt acaccgtgac ggctgaccag 7800
atggcgcgcg agcatcgcgc caagggtgcc ggtgccgccg gtgatcacta cggtgccgtc 7860
ggggtcgaac gccgtgggcg tggtgtccgg gtcggctgcc tcggcggcgg gaacgggaac 7920
acgcccgggg acggggacgg acgccggaac gcgggtcagc cggggcacca ggtgccggtt 7980
cgcacgcacc gcgatctgcg gctcaccaca ggcgagagcc gccggaacga cggtcggtgg 8040
cacttccccg ccgtcgaggt ccgggtcggg gtcgaggtcc agcaggacga accggccggg 8100
atgttccgac tgcgcggagc gcaccatgcc gcacaccgcc gcgcccgcca ggtccgccac 8160
gtcctccccc ggagtggtgg ccaccgcacc acgggtcagt acgaccatcc ggctcgaggc 8220
cggcctctcg tccgccagcc actcctgcag cagacccagg atgcgccggg tgagggtcct 8280
tacgtcctcc ggcacggcgg tggcgcgcgc atcgcggcgg ccgcagtcga ccacgaccgc 8340
atcgggcgcc ggggcccccg cctccagcac tgcggcggcc aggtcggcga ggttcgcgta 8400
cgcgtctgcc gaggggaggc ggtcgctggg ccccaccgtg gccaggcgtt gcggaacgga 8460
cgacgaggtc ggcgccaggg cggtcaccca ctcgaccgcg aagagcgagt cccgccgcgc 8520
cgtaccggag gccgccagca actcctgcgt cgacaccggg cggaccgcga gcgatgcgat 8580
cgaggccacc ggccgcccct cgccgtcgac caccgcgacc gaccatgcgt gcgtgtcccg 8640
cgcctggccg tccggggtga ggcgcacccg cagggtcgac gctccgggcc cgtgcagcct 8700
caccccgctg aaggagaacg gcagggcgcc gtgcgggagt ttgtcggccg ggcggaaggc 8760
cgcggcgtgc agtgcggcgt ccagcagggc ggggtgtacg gagaactggg aggcgtcgcc 8820
gtccgccgtg tcgggaaggc gcacctcggc gaagatgtcc tcgccacgtc gccaggccgc 8880
ccgcaggcct cggaacaccg ggccgtactc gaatccccgc gccgcgtacc cgtcgtacag 8940
gccctccaga tcgtctgcgg tgaacgcgac agcgccctgc ggtggccaga ccgccgccgc 9000
cgctccgtcg ccttgtgggt cggtgagccg tgcgatggct tcatcctggt cggccggtgc 9060
ggggccgacc cggcccgtgg catggcgcac ccacgtggcg tcctcgtacg taccctcgcg 9120
tcgggaggac accgtcaggc ttcggcaccc gttctcatcc ggtgccgcca cccatgcctg 9180
cacctgtact gctccctctt ccgggatgaa cagcggtgat tggagggtga gttcttcgat 9240
gcgccagcag tccacacgat ggcccgcctg aagggcgatc tcgagaaggg cagttccagg 9300
gaggacaacc gcgccggcga tggtgtgatc ggcgagccag gggtgggtgg acagggcgag 9360
ccgaccggtg agcagatagc cctccccctc ggccaccgcg accgccgcgc ccagcaacgg 9420
gtgatgtgcc gggtccaggc cgacggccgc cgcatcccct gtctgggtgg gggtgttgag 9480
ccagtagtgg tggtgttgga aggggtaggt gggtaggtcg atgtgttggg gggttggggt 9540
gtggtgttgg tgggtggtgt gccagttgat ggggtggccg gtggtgtggg cgtgggcgag 9600
ggcgtggagg atttggtggg tgtcgttgtg gttgcggcgg agtgtgctga tgacggtggt 9660
gggggtgtgg gtggtgtcgg tggtttcttg gatggcgggg gtgagggtgg ggtgggggct 9720
gatttcgatg aaggtgtggt ggccttggtg ggtgaggttg gtgatggcgt cggcgaggtg 9780
gacgggttgg cggaggtttc ggtaccagta gtgggcgttg agggtggtgg tgtcggtcca 9840
ttggttgtcg gtggtggagt agaagggggt ggtggcggcc cgtggggtga tgccccgcaa 9900
cagccgtgac agttcctgtt ccacggcctg gatgtggggg cagtgggagg cgtagtcgac 9960
cgggatccgt ttcgcgtcga cgccttggtc ccggtagtgt tccagcgcct gtgtgagagc 10020
ttccgcgttg ccggagatca ccgtggcggt aggggcgttg acggcggcca cccacagccg 10080
gtccggccac atggtctgca gcagcccggt gacctggtcc gccggcagtg gtacggaggc 10140
catgccgccc gaaccgcgca gggtctgcag ggcccggctg cgcagcgcga cggttttcgc 10200
ggcgtcctcc aggctcagcg ctccgctgat gtgggcggcg gtgatttcgc cttgggagtg 10260
gccgatgacg gcgtcggggt taaggccgtg ggattgccag agggcggcga gggaggtcat 10320
gagggcgaag aggacgggct ggatgacgtc ggcgcgctgc cacagggggt cggtgtgggg 10380
ccggtgcagg atgtcggtca gggtccagtc ggtgtggggg gcgagggctt gttcgcaggc 10440
ctggatcgat tgggcgaaga cgggtgaggt cttaagcagc cgcagtccca tcccggccca 10500
ctgaccgccc tgcccgggga acacgaacac caccttgccc tggtcaccgg ccttcctcgc 10560
gatccccgtg acgacggcgg tgtgggcctc gtcggccgcc agagcatcca ggccacccag 10620
gaactccgct cggtcggagc cgatgaccac agcacgatgc tcgaaagagg cgcggcccgt 10680
cgcgagggtg tggccgacgt cggcggggtt cgcctcgggg gcgttggtca ggtacgtgtg 10740
cagttggcgc gcctgggcac gtagcgcttg cgggctcttc gcggagatcg cccagggcag 10800
tgggcctggc gcggtgttgg gctctgccgg ttcgtgggtc tccgcggcag gcggtgcttg 10860
ttccaggatg acgtgggcgt tggtaccgct gacaccgaaa gaggagacgc ccgcacggcg 10920
catccgcccc tgctcgggcc aggccaccga ctcggtcagc aaccgcaccg tcccgcccga 10980
ccagtccaca tgaggtgagg gctcctcggc gtgcagggtc tgcgggagca gcccgtggcg 11040
catggccatc accatcttga tgacgccgcc cacccccgcg gcggcctggg tgtgtccgac 11100
gttggacttg atcgatccca accacaacgg ccgatcggcc gaccgtccct gtccgtacgt 11160
ccccagcagc gcctgggcct cgatggggtc accgagcgtc gtccccgtgc cgtgtccctc 11220
caccgcgtcg atgtccatcg cggccacacc cgcgtttgcc cacgcctggc ggatcacccg 11280
ctcttgcgac gggccgttcg gcgccgtcag cccattgctc gccccgtcct ggttgaccgc 11340
actcccccgt accaccgcca ggacccggtg gccgttgcgc cgggcgtccg aaagccgttc 11400
caccaggagc atccccgccc cctccgccca gcctgttccg tccgcgccgg ccccgaagga 11460
cttgcaccgc ccgtccgggg ccagtccccg ctggcgcgag aactcgacaa aagtggtggg 11520
ggtggacatg acggtgaccc cgccggccag cgccatggag cactcccctg accgcagcgc 11580
ctggcaggcc agatgcaggg cgaccaggga cgacgaacac gccgtgtcca cggtgaccgc 11640
cggcccctcc aggccgaagg tgtaggcgac acgtcccgac gcgacgctgc ccgcgtttcc 11700
attgccgata tagccctcgt agccctctgg gatctcgcgc aggcgggtgc cgtagtcgtg 11760
gtacatcacc ccagcgaaca ccccggtcga actcccccgc agcgacgccg ggtcgatccc 11820
cgcacgttcc agcgcctccc aggagatctc cagcagcagc cgctgctgtg gatccatggc 11880
taccgcctca cgcgggccga tcccgaagaa tgccgcgtcg aactcacccg cgtcgtaaag 11940
gaacccgcct tgccgggtgt aggaggtgcc ggttcggtcc gggtcggggt cgtagagccg 12000
ttccacctcc cagcctcggt ctgcggggaa gtcacccacc gcgtccatac cggaggcgac 12060
cagatcccac aggccctcgg ccgaccgcac acctcccgga aaccggcatg ccatgccgac 12120
gatggccacc ggctcgccgg ctgccccgcc gacggccgca acgggtgggg cggccgtggc 12180
cgccgacgta ccgccggcgc cggccaactc gtcacacagg aaacgggcga gcacgtccgg 12240
acgcggccgg tcgaagatca agccggccgg caggcgcagc ccggtggcct gctgcaaccg 12300
gttccgcagg tccaccgcgg cgagggagtc gaaccccagg tcgcggaacg ccgagtccgc 12360
cgggaccgat tcggccgtgc cgtgccccag caccgcagcc gcccgggagc gcaccagccg 12420
cagcacctcg cgtagccgtt cttccggccc gaggccggtc aaccgccgcc gcagggaggt 12480
cggctcaccg gccgaggcgt cgcgggtccg ggacggccgg gccggagcga cacccgtcag 12540
gtggcgcagg aatggcggga cggatccgga tgtctgaagc tccctcgtgc tgagtgcggc 12600
gggcaccacc agggcgtgtc cacagctcag ggccttgtcg aagagctcca ggccctcgct 12660
gatggccagc ggcctcaggc ccgagcggcg gagccgcgcg tggtccgtgg cctccaactg 12720
cccggtcatg ccctcggtct gctcccaaac tccccaggcc aaggacaccg ccggcagccg 12780
agcggccctc cggtgactcg ccagcgcgtc cagggctgcg ttggcggcgg cgtagttgcc 12840
ctgccctgcc gaacccatca ccccggccag cgaggagaag agcacaaacg cggacagcgg 12900
caggtcgcgt gtgagctggt ggagattgag cgccgaatcg accttggccc gcaggacatg 12960
ggtcaggccc tccggtgtga gggatgtgat caccccgtcg tccagtacgc ctgccgcatg 13020
caccacagcg gtcagcggat ggtcgacgga gatctccgac aatacgttcg caagttgcct 13080
ttggtctgcg gcatcgcacg cggtgatcgt cacctgcgcc ccgagctccg caagttcgac 13140
ccccagtcgc ctggcccccg gggcgtcgag gccacgtcgg ctgatcagca tcagatgccg 13200
tacgccatgg gtggccacca agtggcgggc aagcatggcg ccgaggctgc cggtgccgcc 13260
ggtgatgagt acggtgccgt cctgatccca gggatgctcg tccatggcct cggtggcggt 13320
gccggccagc gggctcagcc ggggtgtgcg cagggctcca tcgcgtacgg ccacttccgg 13380
ttccccgcag gccagggcca tcgacagcat tcggtcgtgg tctttcgcgt cagcgggacg 13440
gtcgtccagg tcgagcagga cgaatcggtc agggtgctcc gactgggcgg agcggatcag 13500
accccgcgcc gccgctcccg ccaggtcttc tacctcctcg tccggcgcag tggccaccgc 13560
ggctcgggtg aggatcacca ggcgactgtc ggcgaaccgg ctgtccgcca accactgctg 13620
cagtaggccc agcatgtggt gtgtggcctg gcgcagcgcc tcactcgact cgtcttcgct 13680
cccggctggg gaccaggcca cgaccgtccg gggaacgggc gcgtcatcgg cgattgccac 13740
gcacagcgcg gccaggtcgg cgtacgtccg ctccggcgga gtgacgaagt cctcgaccgg 13800
gcgcagcgtc ggcccggtcc cgatcatcac ccatggggcg gcttcttcgg ccgctgaagg 13860
ctgggggacg ggttcccatc gcacctcgaa gagcgagcca tggggggcaa ccggggcggc 13920
tgtgcgcaac tcggccgtat gcaactcggc cctggtcacc ggccgcaggg ccaacgaccg 13980
gaccgatgct acgggctcac ccgcctcgtc ggacacgtcg accgacaccg cccccgggcc 14040
atggccgtcc ggcctcagcc ggacgcgcag agccgtcgcc ccggatgcac gcagggtgac 14100
tcctgtccac gtgaacggca gcccgctgtc ctgttgtccc cgtagggagg gcaaaagtgg 14160
aatggtgtgc agtgcggaat ccagcagcgc ggggtgcagc gcgaaccgag ccgcagccgt 14220
gtcgcccagc tgatcctctg acatatacgc ggtcgcgaag acctcgtccc cgcgttgcca 14280
ggcggcgtcc aggcaacgga aggccgcacc gtagtggaaa cccccctcgg ccagttgctc 14340
gtacagtccc tccagatcca cggcgaccgc acccggcggc ggccatgcgc ccgcgggcat 14400
gacaccatcc gccgagtcac cgcagtctgg gtcaggcttg tcggcaaggt acccggcggc 14460
atggcgcgtc cacggctcgc cagccgaatc ggcggtgagg ccgccgacat cgattcgcga 14520
gtagatatcc accgcccggt ttcctgcttc gttcggcgcg gccacagtga cctgcagcac 14580
tacgccggcg ctatccggta cgacgagcgg ggtgtggagc gtaagctcct cgacccgacg 14640
gcagtcggca tgggcggcgg cttggagggc gagttcgaga aaggccgtac cggggagcaa 14700
cacggcgccg ccgacggcgt ggtcggccag ccagggatgc gtacggagag aaatccgccc 14760
agtgaacagc aacccctcac catcggccaa cccgaccgcc gcgcccaaca ggggatgccg 14820
cgccggctca aggcctgcag ccgacatatc accggtggca gtggtggtgt tgagccagta 14880
gtggtggtgt tggaaggggt aggtggggag gtttgggtgg tagtgggtgg gggtgtgatg 14940
gtggtgccag gtggtggggt ggccgtgggt gtggaggtgg gcgagggcgg tgtgggtggt 15000
gtgggtctcg gattggtcgg ggtggagaag cgggacggct agcggggtgt ggtcggggag 15060
gttgtggtgg gtgagggcgg tgagggtgtg gtcggggccg agttcgaggt aggcggtgac 15120
gccgtgggtg tggagttgtt gggtggtgtg ggcgtagtgg accgtctcgc gcgcttgccg 15180
gacccagtag tggggtgtgg tggggtcgtg gtcggtgggg ttggtggtga tgaggggggt 15240
gtggggtggg tggtaggtga gggtttctgc cgtgttgcgg agttcgttga ggagtgggtc 15300
gatgtggtgt gagtggaaag cgccgctgac cttcagcgca gtggtcttgc gtcccatggc 15360
gcggaggttt tcggccgtag cgtctacgtc gtggcgatca ccgctgatga ccacggaccc 15420
gggcgcgttc acggccgcga gtgagacccg tccttcgcat cgtgtgaggt gttcgtggag 15480
ttcgtcgggg tctgcttgga gggtggtcat ggcgccggtg gcggggagag attgcatgag 15540
gcgggcgcgg gtggtgacca ggcgggccgc gtcggggagg gtgaggatcc cggcgaggtg 15600
ggccgcggtg atctctccga gggaatggcc ggcgtagtag tggggggtaa ggccgtggtg 15660
ttcggtgacc aggcggtgca gggcgatctg gagggcgaac agggcgggct gggcgtagcg 15720
ggtctggtgg agcagctggg cggtgtcggt gcccggctcg gcgaacatga catcccgcag 15780
gggatggtcg aggtgtgggt ccaggtgtgc gcaggcttcg tccatggctg cggcgaacag 15840
cgggaaggcg tagtacagcc cgtgtcccat acggggacgt tgcgttccct gcccggaaca 15900
cacaaacgcc agcttccccg cacccgtcgc cacgccccgt accagcccgg cgtgttcctc 15960
accggcctcc agagctccca gctcttcgac gaaggcttcg cggtccgggc ccaggagcac 16020
ggcacggtgg ccgaaggccg tgcggccgga ggccagcgca tacccgacat cagccagccc 16080
cagctcagga tgtgccgcca gatacgcgcg caactgtgct gcctgcgcct gtagtgcctg 16140
ttcggacttg gccgacacca tccacggaaa cgacccggac gagacgtccg ccggatcgtc 16200
gtgctcctca ggcggcgcct gttcgaggat gatgtgcgcg ttggtgccac tgaccccgaa 16260
cgacgacacc cccgcacgcc gcgcacgccc cgtctccggc cacggcaccg cctccgtcag 16320
cagccgcacc ccgcccgagg accagtccac atgcggcgac ggctcctgca catgcaacgt 16380
ccgcggcaac accccatacc gcatcgccat caccatcttg atgacacccg caacccccgc 16440
cgcggcctgc gcatgcccga tattcgactt caccgacccc aaccacaacg ggccgtcccc 16500
cacccgctcc tgaccgtagg tcgccagcaa cgcctgcgcc tcgatcggat cacccagccg 16560
cgtccccgta ccgtgcccct ccaccacatc cacatccgcc ccgccaaccc gcgcactggc 16620
caacgcctgc cggatcaccc gctgctgcga cggaccattc ggcgccgtca aaccattcga 16680
cgcaccatcc tgattcaccg cagaaccccg caccaccgcc aacacccgat gccccaaccg 16740
ccgcgcatca gacaaccgct ccaccaacag catccccacc ccctccgccc accccgtacc 16800
atccgcaccc accccaaacg ccttgcaccg cccatccacc gacaaccccc gctgccgcgc 16860
aaactccacg aaggcaccag gcgtcgccat caccgtcacc ccgccggcca gcgccatgga 16920
gcactccccc gcacgcagcg cctgacacgc cagatgcaac gccaccagag acgacgaaca 16980
cgccgtgtcc accgacaccg cggggccttc caggccgaag gtgtaggcga tacggcccga 17040
ggcgacgctg ccggagtttc cggtgaggac atagccctcg acctcggcca cggtgtcttg 17100
cagccggggg ccgtagtccg tcggcatcac accggcgaag actccggtcc ggctccgccg 17160
taccgacgag gggtcgattc ccgcgtgttc gaaaacctcc caggaggtct ccagcagcag 17220
ccgctgctgc ggatccatcg ccagcgcctc acgtgggccg atgccgaaga atcccgcgtc 17280
gaacttgccc gcgtcgtgga ggaagccccc gtggcgggtg tatgtggtgc ccggcacgtc 17340
ctggtcctgg ggggcgtaga ggcggtcgag gtcccagccg cggtcgctgg ggaatccgct 17400
catggcgtcg gtgccggagg cgaccagatc ccacagcccc tcggccgacc gcacgccccc 17460
cggataccgg caggccatgc cgatgatcgc gatggcctca tccgccacca tcgccacctt 17520
gctcggagca ggagccgcag ccagggtgac cgtatcggcg ccgccgatga ggccgtcccg 17580
caggtgctgg gcgagagcgg caggagtggg atggtcgaag acgatggtgg cggggagccg 17640
caggccggtt gcctccttga ggcggttgcg gagctccacg gcggtgaggg agtcgaagcc 17700
gaggtcacgg aaggccgcct cgcgggcgat ggcgtcgggg ctcgggtggc cgaggaccgt 17760
ggcggcgtgg ccgcggacga gggtgagcag gacctcgtgc tgtgcggcgg ggtcgagccc 17820
ggcgagccgg gctcggaggg aggagcggcc gtcgggtgtc gcggggcgtg gccggtggcc 17880
ggtggtgggg cgtacgaggc tctggaacag gggcggtgcg gtggtgcccc gtagcgcctt 17940
ggtgtcgata cgggccggga gcagcagcgg tgggccgtcg acgagagcgt tgtcgaacag 18000
cgccagggcc tcgtccgtgg ccagtggccg gaggccgacg cgggccatgc gggtgcggtc 18060
ggtctggtcg aggtggccgg tcattccgct ggcctcctcc cacagtcccc atgccaggga 18120
cacggcgggg agtcccatcg cccgtcgctg gaaggcgagc gcgtcgagaa aggcgttggc 18180
ggcggcgtag ttgccctgcc ccggcgagcc caggacaccc gcggcggacg agaacagcac 18240
gaacgcggcc agatccagcc ggcaggtcag gtcgtgcaga tgccaggcgg catcggcctt 18300
cgcgggcaga acggtgtcga ggtggcgcgc ggtgagggag ccgagcgtgg cgtcgtcgag 18360
ggtgcccgcg gcgtggacga cggcggtcag cgggtgctcg gcggggatcc gcccgagcag 18420
ttcggcgagc gcggcacggt cgccgacgtc gcaggtggcg acggtcgccg tggcgcccaa 18480
ctcgtcaatc tcagcgatga gttcgggagc gcctggcgca tcgaggccac ggcggctgat 18540
cagtagcaga tgccgtgctc ggccggtggc ggcgagatgg cgggcgagcg cggcgccgag 18600
ggtaccggtg ccaccggtga tgaggacggt gccgtcgggg tcgggggtgc ggtccagcgt 18660
gagcacattc tttccgacgg tgcggccctg ctggagttgg cggaaggcat ccctggcctg 18720
ccggatgtcc cagcaggtga gcggcagcgg ggtgagcacg ccctcgtcga acagccgcaa 18780
cacggtgcgc aggatctccc cgatgcgctc gggtccggcc tccatcaggt cgtacgcccg 18840
gtaggcgaca ccggggtacg cctgcgccac ctcctccggg tcgcggaggt cggtcttgcc 18900
catctccagg aaccggccgc tgtcgccgga caccaggcgc agcgaggcgt cgacgtactc 18960
atgggccagg gagttgagga cgacgtcgat gttccggccg gtgaaccggc tctcgaagtc 19020
gagggtgcgg gacgaggcga tgcgctccgg ggggatgccg tgggcgcgta gggtgtccca 19080
tttgccgggg ctcgccgtgg cgtagacctc agcgcccagg tggcgggcca gttggacagc 19140
ggccagtccc actccgccgg ccgcagcgtg caccagcacc gactcacccg ggcgcagtcc 19200
tgccaggtcc accagcccgt aataggcggt ggcgaacacg acggggacgg cggcggcctg 19260
gccgaaggtc cagtggtcgg ggacgcggat cacggtggcc cggtcggcga tcgcgaacgg 19320
gccgaacgca tccgcgaaca cgcccatcac ccggtccccg ggcgccaggt cggccacttc 19380
aggcccggct tcgagcacca ccccggcgcc ctcgctgccc agcccccgct gcccagggac 19440
catccccagg gccaccagga catcgcggaa gttcagcccg gccgcacgca cctcgatccg 19500
tacctggtgc gggcccagcg ggtcgcggtc ggccgaggac gccacaggcg ccagcccctg 19560
gaccgtgccc gcgtcggcgc agtcgagccg ccagccgccg ctcccctgtg gcggcgtgag 19620
cagggcgccc ccctcggtgt cggctcgaac cagccgggcc atgtgcggct ggccggcccg 19680
gatggccatc tgcggttcgc ccagcgcgac cgcccgtggt agcgccgccc acgactcctc 19740
ggccgcgtcc aggtccacca ggaggatacg gtcgggctcc tcggcctgcg cggatcgcac 19800
cagtccgcag atcgccgcgc ccgccaggtc catcactccc tccgccggcc cggtggccac 19860
cgcgccgcag gtgaccagca ccagcctcgc gccggccagc cgcgcttcgc cgagccagcc 19920
ctgaagcagg tccagcgctt gtcgggtacg gacacgggta cgggcggtgt ggacgtcggc 19980
gaccgtcgtg tcgtcgggcc cggcggcagc cggagggcag cccacgaaca ccaggtccgg 20040
aggcggatcg cccgcgtcca gggccgtcac cagtccggca aggtccgggt accgccgtac 20100
gggccccgct gccgccgtct ccagcgcggc agcccatcca ggaatggctt ggccaatcac 20160
agcgcaccgc ccgagggccg tcgccggggg ctcgggcgcg gcggtccact ccagggcgta 20220
gagggtgtcg cccgcctggc cgccgagggc ccgcaggtcc tccagggcca ccggccgcag 20280
cgtgagcgat tcggccgagg ccaccggcgc gcccgtctcg tcggccacgt cgacggcaat 20340
cgagtccggg ccctgcggcc gcagccgtac gcgcagcgtc ggtgtgcccg gggagcgcag 20400
gctgacgccg cgccaggcga acggcagcag gccatgtccc gctggattct cctcgcttcg 20460
gccgctgtcg agtgccagcg ggtggagcgc ggcgtcgagc agcgcggggt ggaggcagaa 20520
gcgtggggcg tcggcggcca cctggtcggg gagccgcacc tcggcgtagg tctcctcgcc 20580
ttgccgccag acggcgcgca gcccctggaa ggccgggccg tagccgaagt ctgcggcggc 20640
cagcgtgtcg tagaggccgc tgatgtccag gggctccgcg cccggcggag gccacgcccc 20700
cgccagggtc ccggcaccgt cggcaccgct gatgccatca gtcggggcct cggccgccag 20760
tgtgccggtc gcatgccgtg tccacgcgcc ccgggccgcc ccctcccacg cggggtcgtc 20820
gtcgcgctgt gcgtagacgc tcagtgcgcg gcggccggtc tcgtccggtg ccccgacggc 20880
cacctggagc gccacgccgc cggtgtcggg gaggacgagc ggggcgtgca gggtgagctc 20940
ctccagccgg gggcagccga ggtgtccgcc gacgtgcacg gccagttcca cgtatgcggt 21000
gcccggcagc aggacggtgc cggcgaccgc gtggtcggtg agccaggggt gggtgtgcgg 21060
ggcgagccgg ccggtgagca gatatccgtc gccctcggcg agtccgaccg ccgcgccgag 21120
cagtgggtgt ccgaccgggg ccagccccgc ggccgtcacg tccccggtgg tcgccggggc 21180
gtccagccag tagcggcggc gctggaaggc gtaggtgggc agatctaggt ggtggttgtg 21240
ggggtgggtg tggtggtggg tgtagtggtg gtgccaggtg gtgggtgtgc cgatgccggt 21300
ggtgtgggtg tgggcgaggg cggtgaggaa gcggcgggtg tcgttgtcgc cgcggcggag 21360
gctgccgatc gcggtgacgt cttcggcggt gtcttcggtg gtgtcttcga tggcggggac 21420
gagggtgggg tgggggctga cttcgacgaa gacgcggtgt ccgtcatccg ccagggcctg 21480
gacggcatcg ctgaaacgga caggctggtg caggttgcgg taccagtagg cggcgtccag 21540
ggttgtggtg tccagccagg tgccctccac cgtggagaag aacggcacgc cggacggctg 21600
cgggctgatg tcccccagca gctccagcaa ctcctcccgc aggggctgca catgggggca 21660
gtgcgaggca tagtcgaccg ggatccgccg ggcccgcacc ccggtgccgg cacagtacgc 21720
cagcacctcg tccaccgcct cggcatcccc cgagacggcg gtggagcggg ggccgttgac 21780
cgccgccacc cacaaccgcc ccgcccaccg ctcaccaatg agctgctcca cctcctgggc 21840
aggcagcggc actgaggcca tgccgccccg gccccgcaca gcggccagcg cccggctgcg 21900
cagcgcaaca gtcttcgccg cgtccttcag gctcagcgcc ccacacacat gcgcggccgc 21960
gatctcgccc tgggaatggc caaggaccgc gtcgggttcg ataccgtagg aacgccacag 22020
agcagccaaa gacaccatga cgctgaacag cacaggctgg accacatcgg cccgctccca 22080
caccgcatcc cccgcgtccc ggcgcaggat gtccaccaca gaccagtcca cccacggcgc 22140
cagagcctcc tcgcacgcct gcatccgccg ggcgaacacc ggagaggagg cgagcagacg 22200
cacacccatc ccggcccact gcccaccctg tccgggaaac acgaagacga caccgccccg 22260
gtcaccaccc ggcgcatgac ccgtggtcac ccgccgatcc ggctcacccg ccgccagcgc 22320
ccccaaccct tgcaccagct cctcacggtc cgcggccagg acgaccgcac gatgctccag 22380
cacagcccgc ccacaggcca gacccgcacc cacatcggca agcgaaacgt aagcttggca 22440
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 22500
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 22560
ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt 22620
acgcatctgt gcggtatttc acaccgcata aattccccaa tgtcaagcac ttccggaatc 22680
gggagcgcgg ccgatgcaaa gtgccgatca acataacgat ctttgtagaa accatcggcg 22740
cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat 22800
tcttcgccct gcgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac 22860
ttctcgacag acgtagat 22878
Claims (19)
- 스트렙토마이세스 아베르미틸리스(Streptomyces avermitilis) 균주에서,
아베멕틴 생합성을 수행하는 폴리케타이드 합성효소의 1번 유전자(aveA1)가 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 1번 유전자(milA1)로 치환; 및
아베멕틴 생합성을 수행하는 폴리케타이드 합성효소의 3번 유전자(aveA3) 또는 이의 일부가 밀베마이신 생산 균주에서 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 3번 유전자(milA3) 또는 이의 일부로 치환되고,
상기 aveA3 유전자의 일부가 밀베마이신 생산 균주의 milA3 유전자의 일부의 치환된 것은, aveA3 유전자 중 적어도 모듈 7 코딩 유전자 또는 모듈 7의 DH (dehydratase) 도메인 코딩 유전자가 밀베마이신 생산 균주의 milA3 유전자의 모듈 7 코딩 유전자 또는 모듈 7의 DH (dehydratase) 도메인 및 ER (Enoyl reductase) 도메인 코딩 유전자로 치환된 것을 포함하는 것인,
재조합 스트렙토마이세스 아베르미틸리스 균주. - 삭제
- 제1항에 있어서, 상기 aveA3 유전자의 일부가 밀베마이신 생산 균주의 milA3 유전자의 일부로 치환된 것은,
(i) aveA3 유전자의 모듈 7 코딩 유전자의 KS 도메인 코딩 유전자, AT 도메인 코딩 유전자, KR 도메인 코딩 유전자 및 ACP 도메인 코딩 유전자로 이루어진 군에서 선택된 하나 이상의 유전자가 밀베마이신 생산 균주의 milA3 유전자의 모듈 7 코딩 유전자의 KS 도메인 코딩 유전자, AT 도메인 코딩 유전자, KR 도메인 코딩 유전자 및 ACP 도메인 코딩 유전자로 이루어진 군에서 선택된 하나 이상으로 치환된 것;
(ii) aveA3 유전자의 모듈 8 및 모듈 9의 도메인들 중 하나 이상의 코딩 유전자가 밀베마이신 생산 균주의 milA3 유전자의 모듈 8 및 모듈 9의 도메인들 중 하나 이상의 코딩 유전자로의 치환된 것; 또는
(iii) 치환 (i) 및 (ii)의 조합
을 추가로 포함하는 것인,
재조합 스트렙토마이세스 아베르미틸리스 균주. - 제1항 또는 제3항에 있어서, 상기 밀베마이신 생산 균주는 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus), 스트렙토마이세스 시아네오그리세우스 아속 논시아노게누스(Streptomyces cyaneogriseus subsp. Noncyanogenus), 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis), 또는 스트렙토마이세스 빙첸젠시스(Streptomyces bingchenggensis)인, 재조합 스트렙토마이세스 아베르미틸리스 균주.
- 제1항 또는 제3항에 있어서, 상기 재조합 균주는 밀베마이신 A3, 밀베마이신 A4, 밀베마이신 α2, 밀베마이신 α4, 밀베마이신 G, 및 밀베마이신 D로 이루어진 군에서 선택되는 1종 이상의 밀베마이신을 생산하는 것인, 재조합 스트렙토마이세스 아베르미틸리스 균주.
- 제1항에 있어서, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주는 스트렙토마이세스 아베르미틸리스 DBM-03-A 균주(수탁번호 KCTC12890BP)인, 재조합 스트렙토마이세스 아베르미틸리스 균주.
- 제1항에 있어서, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주는 스트렙토마이세스 아베르미틸리스 DBM-03-B 균주(수탁번호 KCTC12891BP)인, 재조합 스트렙토마이세스 아베르미틸리스 균주.
- 제1항에 있어서, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주는 스트렙토마이세스 아베르미틸리스 DBM-03-C 균주(수탁번호 KCTC13083BP)인, 재조합 스트렙토마이세스 아베르미틸리스 균주.
- 스트렙토마이세스 아베르미틸리스 균주에서, aveA1 유전자가 밀베마이신 생산 균주의 milA1로 치환되고,
상기 밀베마이신 생산 균주는 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus), 스트렙토마이세스 시아네오그리세우스 아속 논시아노게누스(Streptomyces cyaneogriseus subsp. Noncyanogenus), 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis), 또는 스트렙토마이세스 빙첸젠시스(Streptomyces bingchenggensis)이며,
22,23-디하이드로-25-이소프로필-아베멕틴(22,23-dihydro-25-isopropyl-avermectin), 5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin), 5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin) 및 5-메톡시-22,23-디하이드로-25-이소프로필-아베멕틴(5-methoxy-22,23-dihydro-25-isopropyl-avermectin)으로 이루어진 군에서 선택되는 1종 이상의 아베멕틴 유도체를 생산하는,
재조합 스트렙토마이세스 아베르미틸리스 균주. - 제9항에 있어서, 상기 재조합 스트렙토마이세스 아베르미틸리스 균주는, 스트렙토마이세스 아베르미틸리스 DBM-01 균주(수탁번호 KCTC12889BP)인, 재조합 스트렙토마이세스 아베르미틸리스 균주.
- (1) 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 1번 유전자(milA1); 및
(2) 밀베마이신 생산 균주의 밀베마이신 생합성을 수행하는 폴리케타이드 합성효소의 3번 유전자(milA3)의 전부 또는 일부
를 포함하고,
상기 milA3의 일부는 적어도 milA3 유전자 중 모듈 7 코딩 유전자 또는 모듈 7의 DH 도메인 코딩 유전자 및 ER 도메인 코딩 유전자를 포함하는 것인,
재조합 벡터. - 삭제
- 제11항에 있어서, 다음으로 이루어진 군에서 선택된 하나 이상을 추가로 포함하는 재조합 벡터:
밀베마이신 생산 균주의 milA3의 모듈 7의 KS 도메인, AT 도메인, KR 도메인, 및 ACP 도메인 중에서 선택된 하나 이상의 코딩 유전자;
밀베마이신 생산 균주의 milA3의 모듈 8의 코딩 유전자 또는 모듈 8의 도메인들 중 하나 이상의 코딩 유전자; 및
밀베마이신 생산 균주의 milA3의 모듈 9의 코딩 유전자 또는 모듈 9의 도메인들 중 하나 이상의 코딩 유전자. - 제11항에 있어서,
밀베마이신 생산 균주의 milA1 유전자; 및
밀베마이신 생산 균주의 milA3 유전자 중의 모듈 7의 DH 도메인부터 모듈 9의 KR 도메인까지의 영역의 코딩 유전자를 포함하는 milA3 유전자의 일부
를 포함하는, 재조합 벡터. - 제11항, 제13항, 및 제14항 중 어느 한 항에 있어서, 상기 밀베마이신 생산 균주는 스트렙토마이세스 밀베마이시니쿠스 (Streptomyces milbemycinicus), 스트렙토마이세스 시아네오그리세우스 아속 논시아노게누스(Streptomyces cyaneogriseus subsp. noncyanogenus), 스트렙토마이세스 난찬젠시스(Streptomyces nanchangensis) 또는 스트렙토마이세스 빙첸젠시스(Streptomyces bingchenggensis)인, 재조합 벡터.
- 제1항, 제3항, 및 제6항 내지 제8항 중 어느 한 항의 재조합 스트렙토마이세스 아베르미틸리스 균주를 포함하는, 밀베마이신 생산용 조성물.
- 제1항, 제3항, 및 제6항 내지 제8항 중 어느 한 항의 재조합 스트렙토마이세스 아베르미틸리스 균주를 배양하는 단계, 및
상기 배양된 균주 또는 균주의 배양물로부터 밀베마이신을 수득하는 단계를 포함하는,
밀베마이신을 생산하는 방법. - 하기에서 선택되는 아베멕틴 유도체 또는 이의 염:
5-메톡시-22,23-디하이드로-25-메틸-아베멕틴(5-methoxy-22,23-dihydro-25-methyl-avermectin), 및
5-메톡시-22,23-디하이드로-25-에틸-아베멕틴(5-methoxy-22,23-dihydro-25-ethyl-avermectin). - 제9항 또는 제10항 중 어느 한 항의 재조합 스트렙토마이세스 아베르미틸리스 균주 또는 상기 균주의 배양물; 또는
제18항의 아베멕틴 유도체 또는 이의 염
를 포함하는, 해충 방제용 조성물.
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