專利名稱：麥迪霉素高產(chǎn)菌株的制作方法
技術(shù)領(lǐng)域：
本發(fā)明涉及提高麥迪霉素產(chǎn)率的麥迪霉素高產(chǎn)菌株以及利用該菌株生產(chǎn)麥迪霉素的方法，所述麥迪霉素是大環(huán)內(nèi)酯抗生素的一個成員。
背景技術(shù)：
大環(huán)內(nèi)酯抗生素是有效抗革蘭氏陽性細菌，支原體，衣原體等的抗細菌劑并且由于其可以口服施用以及毒性較低被劃分為臨床上重要的抗細菌劑。其中，可以市售獲得的16-元環(huán)大環(huán)內(nèi)酯抗生素由于具有難以誘導(dǎo)抗性，與14-元環(huán)大環(huán)內(nèi)酯抗生素相比與其它藥物的相互作用較少而且也較少影響腸道的優(yōu)點，所以在包括亞洲國家的世界范圍內(nèi)被廣泛使用。
麥迪霉素(圖1)是由Streptomyces mycarofaciens(ATCC 21454)以及放線菌的類似種產(chǎn)生的16-元環(huán)大環(huán)內(nèi)酯抗生素。作為其?；苌锏拿讱W卡霉素(miocamycin)(Omoto，S等，J.Antibiot.，29，536(1976)；Yoshida，T，deng，Jpn.J.Antibiot.，35，1462(1982))在臨床上廣泛使用并且由Streptomyces mycarofaciens的發(fā)酵產(chǎn)物產(chǎn)生。此外，由于Streptomyces mycarofaciens不產(chǎn)生白霉素，通過使用該菌株還具有省去經(jīng)純化等步驟除去白霉素的優(yōu)點。
放線菌長久以來在發(fā)酵工業(yè)領(lǐng)域作為抗生素，生理活性物質(zhì)以及類似的次級代謝產(chǎn)物的生產(chǎn)菌株占據(jù)很重要的位置，而且已經(jīng)通過進行多種菌株育種技術(shù)提高了其產(chǎn)率。也已經(jīng)對由Streptomycesmycarofaciens產(chǎn)生麥迪霉素進行了使用多種誘變劑誘變的菌株育種。根據(jù)這種菌株育種方法，優(yōu)點在于可以很方便獲得具有目的表型的菌株，但是不能闡明向基因中引入了何種類型的突變。由于引入了隨機突變，其后在育種中可能連帶引入沒有用(例如，不提高產(chǎn)率)的突變。
從這個觀點來看，可以通過將低產(chǎn)量菌株的基因組序列與高產(chǎn)量菌株的基因組序列進行比較提取有用的突變，并且可以利用重組DNA技術(shù)構(gòu)建只聚集有用突變的高產(chǎn)菌株。
在產(chǎn)生大環(huán)內(nèi)酯抗生素的微生物中，在很多情況下大多數(shù)的大環(huán)內(nèi)酯生物合成基因一同集中在基因組70到80kb的區(qū)域內(nèi)(Donadio，S.等，Science，252,675(1991)；MacNeil，D，J.等，Gene，115，119(1992)；Schwecke，等，proc.Natl.Acad.Sci.，92，7839(1995))。編碼稱作I型聚酮合成酶(此后也稱作PKS)的巨大多功能蛋白的高度同源基因存在于其基因簇的中心。
PKS基因通常由3到5個基因構(gòu)成，并且其蛋白形成由起始子組件和幾個延伸子組件組成的復(fù)合物。其中每一個在其合成的中間向聚酮鏈添加特異性酰基-CoA前體并特異性修飾β-酮基。因此，通過PKS中這些組件的組成和順序確定聚酮結(jié)構(gòu)。所述組件包含幾個結(jié)構(gòu)域，每一個具有特定的功能。
起始子組件包括結(jié)合前體?；孽；d體蛋白(此后稱作ACP)結(jié)構(gòu)域以及催化向ACP結(jié)構(gòu)域添加?；孽；D(zhuǎn)移酶(此后稱作AT)結(jié)構(gòu)域。根據(jù)這個AT結(jié)構(gòu)域的特異性，確定向其添加的?；?CoA的種類。所有的延伸子組件含有β-酮合成酶(β-酮脂酰?；d體蛋白合成酶，此后也稱作KS)結(jié)構(gòu)域以及AT結(jié)構(gòu)域和ACP結(jié)構(gòu)域，所述β-酮合成酶結(jié)構(gòu)域通過脫羧化縮合向先前存在的聚酮鏈添加新的?；?ACP。
而且，每一延伸子組件含有一些修飾特異性β-酮基的結(jié)構(gòu)域，除此之外，基于所含有的結(jié)構(gòu)域的結(jié)構(gòu)確定β-酮基的修飾。這些包括將β-酮基還原成羥基的β-酮還原酶(此后也稱作KR)結(jié)構(gòu)域，除去羥基并形成雙鍵的脫水酶(此后也稱作DH)結(jié)構(gòu)域以及形成飽和碳鍵的烯?；€原酶(此后也稱作ER)結(jié)構(gòu)域。
最后的延伸子組件通過其從PKS的環(huán)化釋放聚酮由硫酯酶(此后也稱作TE)結(jié)構(gòu)域完成。基于已知的PKS基因的序列信息可以估計PKS的組件，結(jié)構(gòu)域和開放閱讀框架(此后也稱作ORF)的界限。
由PKS形成的聚酮主鏈經(jīng)歷甲基化，?；趸?，還原，特異性糖加成等其它的修飾并最終合成大環(huán)內(nèi)酯抗生素。這些修飾所需的大多數(shù)基因存在于PKS基因的周圍。
編碼脫氧糖生物合成酶的基因已經(jīng)由紅霉素，泰樂霉素等(Sumers，R.G.等，Microbiology，143，3251(1997)；Gaisser，S，等，Mol.Gen.Genet.，256，239(1997)；Merson-Davies，L.A.和Cundliffe，E，Mol.Microbiol.，13，349(1994))得以揭示。這些脫氧糖的合成包括通過添加核苷二磷酸活化葡萄糖以及隨后的脫氧化，還原，表異構(gòu)化，胺化，甲基化等反應(yīng)。這些糖通過活化特異性糖基轉(zhuǎn)移酶被引入大環(huán)內(nèi)酯。
由于麥迪霉素的結(jié)構(gòu)與泰樂霉素的結(jié)構(gòu)類似，因此人們認為它們經(jīng)歷幾乎相同的生物合成途徑。麥迪霉素的生物合成起始于作為聚酮主鏈前體的丙二?；?CoA，甲基丙二?；?CoA，乙基丙二酰基-CoA以及甲氧基丙二?；?CoA的合成。這些前體通過由聚酮合成酶的逐步縮合反應(yīng)進行環(huán)化，并由此合成聚酮主鏈。隨后經(jīng)糖加成，羥基化，甲酰化，乙?；刃揎椃磻?yīng)最終合成麥迪霉素。
為了通過基因重組技術(shù)提高其產(chǎn)率，已經(jīng)進行編碼限速生物合成反應(yīng)的基因的表達加強，調(diào)節(jié)生物合成基因表達的基因的表達加強或者基因破壞，中斷不必要的次級代謝系統(tǒng)等(Kennedy，J.and Turner，G，Mol.Gen.Genet.，253，189(1996)；ReviewBalts，R.H.，Biotechnology ofAntibiotics Second Edition，Revised and Expanded，Marcel Dekker，Inc.，New York，p.49(1997)；ReviewHutchinson，C.R.and Colombo，A.L.，Ind.Microbiol.Biotechnol.，23，647(1999)；ReviewBrakhage，A.A.，Microbiol.Mol.Biol.Rev.，62，547(1998))。因此，當(dāng)生物合成基因被確定時，可以通過基因重組技術(shù)，通過將其與合適的載體連接并導(dǎo)入次級代謝產(chǎn)物生產(chǎn)菌株提高其產(chǎn)率。

發(fā)明內(nèi)容
上述的方法是針對提高限速基因的表達量或者去除限速基因并由此提高或作為基因產(chǎn)物的酶蛋白的量。然而，很難通過精確掌握生物合成中間體的產(chǎn)率確定限速步驟。例如，有一些不能預(yù)測生物合成流程圖，不能檢測所預(yù)測的生物合成中間體，需要生物合成中間體的標(biāo)準(zhǔn)物等等的情況。
本發(fā)明人已經(jīng)通過利用突變處理的育種從產(chǎn)生麥迪霉素的Streptomyces mycarofaciens獲得了高產(chǎn)率菌株，并將來源于這種高產(chǎn)菌株的麥迪霉素生物合成基因的DNA序列與野生型菌株的序列(低產(chǎn)菌株)進行了比較，并且檢驗發(fā)現(xiàn)了特異于高產(chǎn)菌株的突變，即引起高產(chǎn)的突變。
也就是說，本發(fā)明人已經(jīng)成功分離了一包含麥迪霉素高產(chǎn)菌株-來源的基因組DNA的基因組文庫的麥迪霉素生物合成基因的DNA片段，所述基因組文庫利用Streptomyces mycarofaciens-來源的DNA片段作為探針制備而成，所述探針已經(jīng)利用與放線菌-來源的聚酮合成酶基因同源的序列作為引物通過聚合酶鏈?zhǔn)椒磻?yīng)(在下文也稱作PCR)得以擴增。通過將由此獲得的高產(chǎn)菌株來源的麥迪霉素生物合成基因與野生型菌株來源的麥迪霉素生物合成基因(美國專利公開2004-0091975A1，JP-A-2004-49100)的DNA序列進行比較，人們發(fā)現(xiàn)存在聚酮合成酶基因的β酮合成酶結(jié)構(gòu)域的部分取代并由此提高了麥迪霉素的產(chǎn)率?；谏鲜稣J識，我們已經(jīng)繼續(xù)了深入研究并由此完成了本發(fā)明。
因此，本發(fā)明包含下列實施方案。
1.產(chǎn)麥迪霉素的放線菌，其含有麥迪霉素生物合成基因或其同源物，其中麥迪霉素生物合成基因的聚酮合成酶基因中的至少一個組件或至少一個組件的部分序列被取代以便編碼其它組件的相應(yīng)氨基酸序列。
2.根據(jù)項1的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中的KS3β-酮脂酰?；d體蛋白合成酶的氨基酸序列的核苷酸序列被全部或部分取代以便編碼ORF 1中的KS2β-酮脂酰酰基載體蛋白合成酶的相應(yīng)氨基酸序列。
3.根據(jù)項2的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF 2中KS3的第157至第420位的氨基酸序列的核苷酸序列被部分或全部取代以便編碼ORF 1中KS2的第2705至第2968位的部分或全部的相應(yīng)氨基酸序列。
4.根據(jù)項3的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中KS3的第157至第254位的氨基酸序列的核苷酸序列被部分或全部取代以便編碼ORF 1中KS2的第2705至第2802位的部分或全部的相應(yīng)氨基酸序列。
5.根據(jù)項4的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中KS3的第157至第186位的氨基酸序列的核苷酸序列被部分或全部取代以便編碼ORF 1中KS2的第2705至第2734位的部分或全部的相應(yīng)氨基酸序列。
6.根據(jù)項1到5任一項的產(chǎn)麥迪霉素的放線菌，其中的放線菌為Streptomyces mycarofaciens。
7.根據(jù)項6的產(chǎn)麥迪霉素的放線菌，其中的放線菌為保藏號為FERM BP-10501的Streptomyces mycarofaciens 1149-38菌株。
8.產(chǎn)麥迪霉素的放線菌，其是通過對項1到5任一項所述的產(chǎn)麥迪霉素的放線菌進一步突變獲得的。
9.產(chǎn)生麥迪霉素的方法，其包括培養(yǎng)項1到5任一項所述的產(chǎn)麥迪霉素的放線菌；以及分離產(chǎn)生的麥迪霉素。
10.產(chǎn)生基本上不含白霉素的麥迪霉素的方法，其包括培養(yǎng)項6或7所述的產(chǎn)麥迪霉素的放線菌；以及分離產(chǎn)生的麥迪霉素。
本發(fā)明提供了提高麥迪霉素產(chǎn)率的麥迪霉素高產(chǎn)菌株以及利用該菌株生成麥迪霉素的方法，所述麥迪霉素是大環(huán)內(nèi)酯類抗生素的成員。


圖1是顯示麥迪霉素結(jié)構(gòu)的圖。
圖2是Streptomyces mycarofaciens菌株的分類圖。
圖3是顯示Streptomyces mycarofaciens菌株ATCC 21454的麥迪霉素聚酮合成酶的orf 1存在的KS2和ORF2中的KS3的氨基酸序列和Streptomyces mycarofaciens菌株1149-38的麥迪霉素聚酮合成酶的ORF2中存在的KS3(高KS3)的氨基酸序列比較的相似性。圖中的氨基酸數(shù)目通過將每一orf的起始密碼子定義為1進行表示。
具體實施例方式
以下詳細描述了本發(fā)明。
微生物的保藏獲自Meiji Seika Kaisha的Streptomyces mycarofaciens SF8371149-38已經(jīng)在2005年2月16日保藏在獨立行政法人產(chǎn)業(yè)技術(shù)綜合研究所特許生物保藏中心(International Patent Organism Depositary，NationalInstitute of Advanced Industrial Science and Technology(郵政編碼305-8566；Central 6，1-1-1Higashi，Tsukuba，Ibaraki，日本))。保藏號是FERM P-20405(在2006年2月2日轉(zhuǎn)成FERM BP-10501)。
轉(zhuǎn)化有pCOMW1的大腸桿菌已經(jīng)在2002年7月16日保藏在獨立行政法人產(chǎn)業(yè)技術(shù)綜合研究所特許生物保藏中心(郵政編碼305-8566；Central 6，1-1-1 Higashi，Tsukuba，Ibaraki，日本)。保藏號是FERM-18935(在2002年8月21日轉(zhuǎn)成FERM BP-8168)。
轉(zhuǎn)化有pCOMW2的大腸桿菌已經(jīng)在2002年7月16日保藏在獨立行政法人產(chǎn)業(yè)技術(shù)綜合研究所特許生物保藏中心(郵政編碼305-8566；Central 6，1-1-1 Higashi，Tsukuba，Ibaraki，日本)。保藏號是FERM-18936(在2002年8月21日轉(zhuǎn)成FERM BP-8169)。
轉(zhuǎn)化有pCOMW4的大腸桿菌已經(jīng)在2002年7月16日保藏在獨立行政法人產(chǎn)業(yè)技術(shù)綜合研究所特許生物保藏中心(郵政編碼305-8566；Central 6，1-1-1 Higashi，Tsukuba，Ibaraki，日本)。保藏號是FERM-18937(在2002年8月21日轉(zhuǎn)成FERM BP-8170)。
本發(fā)明的麥迪霉素生物合成基因野生型菌株(Streptomyces mycarofaciens ATCC 21454)-來源的麥迪霉素生物合成基因公開在美國專利2004-0091975A1和JP-A-2004-49100中。
根據(jù)本發(fā)明，麥迪霉素的產(chǎn)率可以通過更換麥迪霉素生物合成基因，聚酮合成酶基因中所含的每一組件或者其部分序列得以提高，由此使其編碼其它組件的相應(yīng)的氨基酸序列。置換序列沒有特殊限制，條件是麥迪霉素的產(chǎn)率得以提高，而且聚酮合成酶(麥迪霉素的產(chǎn)率)的酶活性可以得以提高，例如通過利用上游β酮合成酶結(jié)構(gòu)域或者其部分取代相同的下游結(jié)構(gòu)域的相應(yīng)部分，與麥迪霉素高產(chǎn)菌株-來源的聚酮合成酶的情況類似。
待置換的氨基酸序列的長度沒有特殊限制，但優(yōu)選1到300個殘基，更優(yōu)選1到200個殘基，最優(yōu)選1到100個殘基。
待提高的麥迪霉素產(chǎn)率基于未經(jīng)歷置換的菌株為1.2倍或更多，優(yōu)選1.4倍或更多，麥迪霉素的產(chǎn)率由以下實施例1到8所述的方法進行確定，可以通過將其與其他突變結(jié)合獲得與野生型菌株(ATCC 21454)相比10倍或更多，優(yōu)選15倍或更多，更優(yōu)選18倍或更多倍的提高。
在上述的取代中，人們希望用ORE1上KS2結(jié)構(gòu)域的相應(yīng)部分取代ORF2上KS3結(jié)構(gòu)域的氨基酸序列或者其部分，更希望用KS2結(jié)構(gòu)域(從第2705到第2968位置的部分或者全部氨基酸序列)的相應(yīng)部分取代KS3結(jié)構(gòu)域的第157到第420位置的部分或者全部氨基酸序列，還希望用KS2結(jié)構(gòu)域的相應(yīng)部分(從第2705到第2802位置的部分或者全部氨基酸序列)取代從KS3結(jié)構(gòu)域第157到第254位置的部分或者全部氨基酸序列，進一步希望用KS2結(jié)構(gòu)域的相應(yīng)部分(從第2705到第2734位置的部分或者全部氨基酸序列)取代KS3結(jié)構(gòu)域的從157到第186位置的部分或者全部氨基酸序列。氨基酸的數(shù)目通過將翻譯起始密碼子(atg/Met)定義為1進行計數(shù)。
本發(fā)明的麥迪霉素的制備方法可以例如通過以下方法獲得產(chǎn)生包含如下成分的麥迪霉素的放線菌麥迪霉素生物合成基因或者其同源物，其中麥迪霉素生物合成基因，聚酮合成酶基因，或者其部分序列中所含的一或兩個或更多組件被其它組件的相應(yīng)的氨基酸序列取代。
提高麥迪霉素產(chǎn)率的菌株通過對麥迪霉素生產(chǎn)菌株Streptomycesmycarofaciens施加紫外線輻射或者誘變劑(例如，亞硝基胍)處理獲得。制備這種菌株的麥迪霉素生物合成基因并與已經(jīng)確定的生物合成基因相比較尋找引入突變的位置。為了確定突變的效果，通過同源重組將由此獲得的突變基因轉(zhuǎn)入沒有突變的麥迪霉素生產(chǎn)菌株中(例如，Bierman等人的方法，Gene，116，46(1992))，并驗證產(chǎn)率的改變。
聚酮合成酶具有上述的KS，AT以及ACP結(jié)構(gòu)域，但其酶活性(反應(yīng)速度)被認為是不規(guī)則的，人們因此認為代謝中間產(chǎn)物得以聚集。因此，人們認為聚酮合成能力通過明確低反應(yīng)速度組件，以及用高反應(yīng)速度組件對其進行置換得以提高。示范性的置換方法可以利用基因操作領(lǐng)域所通常使用的方法進行構(gòu)建，其實例包括引入限制性內(nèi)切酶識別序列以及在其中插入目的置換位點，或者通過PCR以及連接擴增目的位點的方法。
根據(jù)本發(fā)明，麥迪霉素產(chǎn)率提高的麥迪霉素生物合成基因除了每一組件或者其部分的上述的氨基酸序列的取代，可以包含一或者多個選自缺失，添加，插入以及取代的突變(在下文具有這種突變的基因稱作“類似物”)，在所述麥迪霉素生物合成基因中對應(yīng)于麥迪霉素生物合成基因，聚酮合成酶基因或者其部分序列中所含的每一組件的氨基酸序列被對應(yīng)于其它組件的氨基酸序列取代。一或者多個選自缺失，插入，添加以及取代的突變的數(shù)目沒有特殊限制，條件是保持麥迪霉素的產(chǎn)率，而且優(yōu)選突變從1到10，更優(yōu)選從1到6，最優(yōu)選從1到4個氨基酸殘基。
可以例如根據(jù)下列方法制備這種進一步添加了突變的菌株。
通過引入隨機突變或者位點特異性突變有可能獲得本發(fā)明天然酶的氨基酸序列中實現(xiàn)缺失，添加，插入以及取代一或兩個或更多氨基酸殘基至少之一的基因。由此，也可以獲得編碼具有本發(fā)明酶活性，但是在諸如最適溫度，穩(wěn)定溫度，最適pH值，穩(wěn)定pH，底物特異性等特性稍有不同的本發(fā)明的酶的基因。
對于引入隨機突變的方法，例如在亞硫酸氫鈉作用下引起胞嘧啶轉(zhuǎn)化成尿嘧啶的轉(zhuǎn)換性突變的方法[Weiher，H.等人，Proceedings of theNational Academy of Sciences of the USA，vol.79，1408-1412(1982)]可用作化學(xué)處理DNA的方法，在合成雙鏈過程中在[α-S]dNTP存在下產(chǎn)生堿基取代的方法[Shiraishi，H.等人，Gene，vol.64，313-319(1988)]可用作生化法，以及通過向反應(yīng)系統(tǒng)添加錳實施PCR降低核苷酸摻入的準(zhǔn)確度的方法[Analytical Biochemistry，vol.224，347-353(1995))可用作采用PCR的方法。
對于引入位點特異性突變的方法，例如可以采用如下方法利用琥珀突變的方法[缺口雙鏈體方法，Nucleic Acids Research，vol.12，no.24，pp.9441-9456(1984)]，利用限制性內(nèi)切酶識別位點的方法[AnalyticalBiochemistry，vol.200，81-88(1992)，Ito，W.等人，Gene，vol.102，pp.67-70(1991)]，利用dut(duTpase)以及ung(尿嘧啶DNA糖基化酶)突變的方法[Kunkel method，Kunkel，A.T.，Proceedings of the NationalAcademy of Sciences of the USA，vol.82，pp.488-492(1985)]，利用使用DNA聚合酶以及DNA連接酶的琥珀突變的方法[寡核苷酸-指導(dǎo)的雙重琥珀(ODA)方法的方法，Hashimoto-Goto，T.等人，Gene，vol.152，pp.271-275(1995)，JP-A-7-289262]，利用誘導(dǎo)DNA修復(fù)系統(tǒng)的宿主的方法(JP-A-8-70874)，利用催化DNA鏈交換反應(yīng)的蛋白的方法(JP-A-8-140685)，利用用于添加限制性內(nèi)切酶識別位點的突變誘導(dǎo)應(yīng)用的兩個引物的PCR方法(USPS,512,463)，利用具有滅活耐藥性基因以及兩個引物的雙鏈DNA載體的PCR方法[Shen，T.等人，Gene，vol.103，pp.73-77(1991)]，利用琥珀突變[WO 98/02535]等的PCR方法。
此外，位點特異性突變可以通過利用市售的試劑盒被輕易地引入。作為市售的試劑盒，可以使用如下的試劑盒例如利用缺口雙鏈體方法的Mutan(注冊商標(biāo))-G(由Takara Bio制造)，利用Kunkel法的Mutan(注冊商標(biāo))-K(由Takara Bio制造)，利用ODA方法的Mutan(注冊商標(biāo))-Express Km(由Takara Bio制造)，利用用于突變誘導(dǎo)應(yīng)用的引物以及Pyrococcus furiosus-來源的DNA聚合酶的QuikChangeTM Site-DirectedMutagenesis Kit(由STRATAGENE制造)等等，并且可以使用作為應(yīng)用PCP方法的試劑盒的如下試劑盒TaKaRa LA PCR體外誘變試劑盒(由Takara Bio制造)，Mutan(注冊商標(biāo))-Super Express Km(由Takara Bio制造)等。
由麥迪霉素產(chǎn)率提高的菌株制備麥迪霉素產(chǎn)率提高的麥迪霉素生物合成基因的方法描述在例如從Streptomyces mycarofaciens菌株1149-38進行制備的例子中。
本發(fā)明的麥迪霉素生物合成基因可以例如通過下列方法分離自Streptomyces mycarofaciens菌株1149-38。因為該序列如本發(fā)明公開中所揭示，可以人工合成相關(guān)的基因，但是可以從Streptomycesmycarofaciens菌株1149-38高效地制備。
基因組DNA通過Kieser，T.等人在Practical Streptomyces Genetics，The John Innes Foundation，Norwick，UK(2000)中描述的常規(guī)方法由Streptomyces mycarofaciens strain 1149-38的細胞提取而來。由Streptomyces mycarofaciens的基因組DNA組成的基因組文庫由用合適的限制性內(nèi)切酶消化基因組DNA然后用合適的載體連接所述產(chǎn)物進行制備。
作為載體，例如可以使用質(zhì)粒載體，噬菌體載體，粘粒載體，BAC載體以及類似的多種載體和質(zhì)粒。
可以通過使用合適的探針雜交由基因組文庫獲得包含目的麥迪霉素生物合成基因的DNA片段的制備物。例如，基于通常已知的聚酮合成酶的氨基酸序列的保守區(qū)域合成合適的引物，利用它們以及利用Streptomyces mycarofaciens的基因組DNA作為模板進行PCR，由此擴增的DNA片段可被用作探針。此外，因為麥迪霉素生物合成基因的序列如本發(fā)明公開所揭示，基于序列信息合成合適的引物，利用Streptomyces mycarofaciens的基因組DNA作為模板進行PCR，由此擴增的DNA片段也可以被用作探針。利用由此獲得的DNA片段作為探針進行基因組文庫的篩選。
此外，因為麥迪霉素生物合成基因的序列如本發(fā)明公開被揭示，基于序列信息合成用于目的基因擴增的引物，利用Streptomycesmycarofaciens的基因組DNA作為模板進行PCR，然后由此擴增的DNA片段可通過將其與合適的載體連接被分離。
由此，對于提高麥迪霉素生物合成能力的聚酮合成酶基因，有可能通過在分離之前直接利用突變株產(chǎn)生麥迪霉素，或者可以將麥迪霉素生物合成能力提高的聚酮合成酶基因轉(zhuǎn)入麥迪霉素生產(chǎn)菌株中，但是也可以通過連同總長度的麥迪霉素生物合成簇轉(zhuǎn)化到任選的宿主中獲得目的麥迪霉素。
應(yīng)宿主種類的需要確定所用的載體，但是沒有特別限制，例如pBR322系統(tǒng)以及pUC系統(tǒng)的載體可以被引作大腸桿菌載體，pUB110系統(tǒng)，pPL603系統(tǒng)以及pC194系統(tǒng)的載體可作為枯草芽胞桿菌載體，pYC系統(tǒng)以及pYE系統(tǒng)的載體作為酵母載體，pIJ101系統(tǒng)，pSET152系統(tǒng)，pSG5系統(tǒng)，pSCP2*系統(tǒng)，SAM2系統(tǒng)，pKC1139系統(tǒng)以及C31系統(tǒng)的載體可作為放線菌載體(Kieser，T.等，Practical Streptomyces Genetics，The John Innes Foundation，Norwick，UK(2000))。
然后，利用由此獲得的質(zhì)粒將基因轉(zhuǎn)入宿主。宿主應(yīng)所用載體的種類的需要可任選自放線菌，大腸桿菌，枯草芽胞桿菌，酵母，絲狀真菌及其他微生物。
當(dāng)載體用于放線菌應(yīng)用時作為宿主的尤其優(yōu)選的實例包括Streptomyces mycarofaciens，Streptomyces coelicolor，吸水鏈霉菌(Streptomyces hydroscopicus)，弗雷德氏鏈霉菌(Streptomycesfradiae)，Streptomyces lividans，Streptomyces kitasatoensis，Streptomycesambofaciens以及Streptomyces themotolerans。
作為將載體轉(zhuǎn)入宿主菌株的方法，最有效方法的選擇取決于宿主以及載體的類型。當(dāng)應(yīng)用放線菌載體時，可以進行結(jié)合大腸桿菌的轉(zhuǎn)移，用放線菌噬菌體感染，向宿主菌株引入原生質(zhì)體等(Kieser，T.等人，Practical Streptomyces Genetics，The John Innes Foundation，Norwick，UK(2000))。由所應(yīng)用的載體具有的遺傳指標(biāo)，諸如抗生素抗性，痘痕(pock)形成，黑色素生物合成等等可被用于選擇由轉(zhuǎn)化獲得的重組體。
通過通常已知的方法培養(yǎng)獲得的重組體，可以檢驗它們新近獲得的特性。
麥迪霉素的產(chǎn)生可以通過培養(yǎng)上述包含提高麥迪霉素生物合成能力的麥迪霉素合成基因，或者轉(zhuǎn)入了提高麥迪霉素生物合成能力的麥迪霉素合成基因的重組體的突變株產(chǎn)生麥迪霉素。
對于介質(zhì)，可以使用通常應(yīng)用的組分諸如葡萄糖，蔗糖，淀粉糖漿，糊精，淀粉，甘油，糖漿，動植物油等作為碳源。同樣，大豆粉，麥芽，玉米漿，棉子粉，肉汁，多聚蛋白胨，麥芽汁，酵母抽提物，硫酸銨，硝酸鈉，尿素等可用作氮源。除了這些組分，需要時也可以添加鈉，鉀，鈣，鎂，鈷，氯，磷酸(磷酸氫二鈉等)，硫酸(硫酸鎂等)及其他可以形成離子的無機鹽。同樣，需要時還可以添加硫胺素(鹽酸硫胺素等等)等多種維生素，谷氨酸(谷氨酸鈉等等)，天門冬酰胺(DL-天門冬酰胺等等)等氨基酸，核苷酸等微量養(yǎng)分以及抗生素等選擇藥物。此外，可以任選添加能夠有助于菌株生長以及促進麥迪霉素產(chǎn)生的有機物以及無機物。
介質(zhì)pH例如為大約從pH 5.5到pH 8。對于培養(yǎng)法，可以采用需氧條件下的固體培養(yǎng)，振蕩培養(yǎng)，有氧攪拌培養(yǎng)或者沉浸有氧培養(yǎng)(submerged aeration culture)，其中沉浸有氧培養(yǎng)尤其合適。適合于培養(yǎng)的溫度為從15℃到40℃，但是在大多數(shù)情況下菌株生長在大約從22℃到30℃的溫度下。雖然麥迪霉素的產(chǎn)量依賴于培養(yǎng)基以及培養(yǎng)條件或者所使用的宿主的不同而不同，通過任何一種培養(yǎng)法通常在2天到10天內(nèi)其累積達到最大。當(dāng)培養(yǎng)期間麥迪霉素的量達到最大時，中止培養(yǎng)并分離目的物質(zhì)并由培養(yǎng)的混合物進行純化。
為了從培養(yǎng)的混合物收集麥迪霉素，可以利用諸如溶劑萃取，離子交換樹脂法，吸附或者分配柱層析，凝膠過濾，滲析，沉淀，結(jié)晶等可單獨或者任選組合使用的特性，由常規(guī)分離方式進行提取和純化。例如，用丙酮，甲醇，丁醇，乙酸乙酯，乙酸丁酯等由培養(yǎng)的混合物提取。
為了更進一步純化麥迪霉素，利用硅膠，氧化鋁等吸附劑，Sephadex LH-20(由Amersham Bioscience制造)，Toyopearl HW-40(由Tosoh制造)等實施層析是非常有效的。
通過這種方法或者其任選組合獲得純的麥迪霉素。
實施例以下基于實施例示范性描述了本發(fā)明，雖然這些并不限制本發(fā)明。
實施例1-1分離Streptomyces mycarofaciens(ATCC 21454)-來源的基因組DNA以及制備基因組文庫將冷凍的Streptomyces mycarofaciens(ATCC 21454)菌種接種到50ml的S#14培養(yǎng)基(2％葡萄糖，1％多聚蛋白胨，0.05％K2HPO4，0.05％MgSO4·7H2O，0.3％NaCl，pH 7.0)中并在28℃培養(yǎng)20小時。利用0.22μm的Bottle top過濾器(由Corning制造)過濾培養(yǎng)液體然后用10mMEDTA洗滌過濾器上的細胞兩次回收細胞。用液氮冷凍由此獲得的細胞然后利用研缽和研杵研磨。利用ISOPLANT(由Nippon Gene制造)并根據(jù)附裝的說明書從研磨的細胞分離基因組DNA。
用Sau3AI部分消化由此分離的基因組DNA，然后對其末端脫去磷酸。用已經(jīng)用BamHI和XbaI(單獨XbaI位點的脫磷酸)消化的SuperCosI(由Stratagene制造)連接這種DNA片段制備重組體粘粒載體。利用MaxPlax Packaging Extract(由Epicentre Technologies制造)并根據(jù)附裝說明書在這種重組體粘粒載體上進行體外包裝。其后，用這種重組噬菌體感染大腸桿菌XL1-Blue MR菌株并在板上培養(yǎng)實現(xiàn)集落的形成。
實施例1-2探針的制備下列引物由PKS基因的保守區(qū)域制備而來。
KS-F5′-CGGTSAAGTCSAACATCGG-3′(SEQ ID NO1)KS-R5′-GCRATCTCRCCCTGCGARTG-3′(SEQ ID NO2)使用KS-F和KS-R，利用基因組DNA作為模板進行PCR。使用ExTaqDNA聚合酶(由Takara Bio制造)進行PCR。使用TOPO TA克隆試劑盒(由Invitrogen制造)并根據(jù)其附裝的說明書將由此擴增的DNA片段插入pCR2.1-TOPO質(zhì)粒載體。
使用DNA測序試劑盒dRhodamine Terminator Cycle SequencingReady Reaction(由Applied Biosystems制造)以及ABI PRISM GeneticAnalyzer(由Applied Biosystems制造)并根據(jù)其附裝的說明書進行所插入DNA片段的測序。由此證實分離的DNA片段是PKS基因的一部分。
實施例1-3粘粒文庫的篩選使用包含一部分麥迪霉素PKS基因作為模板以及使用引物KS-F以及KS-R由PCR擴增DNA片段，并用作雜交的探針。
將Hybond-N+膜(Amersham Pharmacia Biotech)置于形成基因組文庫的菌落的板上，由此實現(xiàn)集落對其的附著力。通過將這種膜進行堿處理，實現(xiàn)細胞溶解并由此同樣將膜上的重組體粘粒DNA變性成單鏈并粘附到膜上。使用ECL指導(dǎo)的核酸標(biāo)記和檢測系統(tǒng)(由AmershamBioscience制造)并根據(jù)其附裝的說明書檢測膜上的陽性克隆。用這種方式，分離包含與探針同源的區(qū)域的粘?？寺COMW1(FERM P-18935)和pCOMW2(FERM P-18936)。經(jīng)PCR由部分分析的pCOMW1(FERMP-18935)末端序列新近制備探針。使用這種探針再次進行基因組文庫的篩選，并分離pCOMW4(FERM P-18937)。
實施例1-4核苷酸序列的測定用HaeIII部分消化克隆pCOMW1(FEPMP-18935)和pCOMW2(FERMP-18936)，然后由電泳純化約2kb的片段并與已經(jīng)用SmaI消化的pUC19連接。將這種質(zhì)粒引入大腸桿菌XL1-blue菌株，由任選的菌落提取質(zhì)粒，使用-21M13正向引物和M13反向引物作為引物由ABI 3700(由Applied Biosystems制造)并根據(jù)其附裝的說明書進行測序。從由此獲得的結(jié)果，將未充分分析的區(qū)域再次由基于已經(jīng)分析的核苷酸序列新設(shè)計的引物進行測序。其后，基于該分析結(jié)果，由引物waking確定pCOMW4(FERM P-18937)的部分序列。
實施例1-5Streptomyces mycarofaciens菌株1251-2麥迪霉素生物合成基因簇的克隆以及序列分析Streptomyces mycarofaciens菌株1251-2是其麥迪霉素產(chǎn)率由亞硝基胍處理得以提高的菌株(圖2)。因為人們認為這種菌株的產(chǎn)率的提高應(yīng)歸于麥迪霉素生物合成基因的突變，所以克隆該基因簇并與ATCC21454-來源的生物合成基因相比較。
與克隆ATCC 21454-來源的基因同樣的方法進行Streptomycesmycarofaciens菌株1251-2來源的基因組DNA的分離，制備粘粒文庫并克隆。
篩選Streptomyces mycarofaciens菌株1251-2-來源的粘粒文庫獲得pCOM1和pCOM2。由其序列分析的結(jié)果，顯示在其中含有麥迪霉素生物合成基因中的orf 36到11。
實施例1-6各菌株之間比較麥迪霉素生物合成基因簇對于以上述方式獲得的2種麥迪霉素生物合成基因簇的各翻譯區(qū)，比較其DNA序列。Streptomyces mycarofaciens菌株ATCC 21454的麥迪霉素聚酮合成酶的orf 1的核苷酸序列顯示在SEQ ID NO3中，其氨基酸序列顯示在SEQ ID NO4中，ORF2的核苷酸序列顯示在SEQ ID NO5中，其氨基酸序列顯示在SEQ ID NO6中，Streptomyces mycarofaciens菌株1251-2的麥迪霉素聚酮合成酶的ORF2的核苷酸序列顯示在SEQ IDNO7中，其氨基酸序列顯示在SEQ ID NO8中。
結(jié)果顯示在Streptomyces mycarofaciens菌株1251-2來源的麥迪霉素生物合成基因簇中，與苷元生物合成有關(guān)的ORF2的KS3結(jié)構(gòu)域被突變并具有與存在于ORF 1中的KS2結(jié)構(gòu)域部分相同的序列(圖3)。
實施例1-7檢測具有突變體麥迪霉素生物合成基因的麥迪霉素高產(chǎn)菌株為了判斷引入上述特異于Streptomyces mycarofaciens菌株1251-2的聚酮合成酶的突變的繼代培養(yǎng)的菌株的階段，通過PCR法進行突變的檢測。使用W-ORF2-U與ORF2-L以及H-ORF2-U與ORF2-L的組合作為引物并使用Takara Bio生產(chǎn)的LA-PCR試劑盒，在94℃熱變性3分鐘然后重復(fù)由94℃1分鐘以及68℃2.5分鐘組成的步驟的25個循環(huán)，由此擴增ORF2的KS3結(jié)構(gòu)域。
W-ORF2-U5′-GTGATGTATGACGACTACGG-3′(SEQ ID No9)H-ORF2-U5′-AAACCTCGGAAGTGTGGTCT-3′(SEQ ID No10)ORF2-L5′-ATCGAGGGCGTCGGCGGTAC-3′(SEQ ID No11)結(jié)果，在菌株938-15和菌株1149-38之間引入了突變。
實施例1-8
比較Streptomyces mycarofaciens菌株938-15和菌株1149-38(FERMBP-10501)的麥迪霉素的產(chǎn)率。
將每一Streptomyces mycarofaciens菌株938-15和菌株1149-38(FERMBP-10501)的0.1ml份的冷凍菌種接種在內(nèi)徑2cm的試管中所含的10ml種子培養(yǎng)基(2％可溶性淀粉，1％葡萄糖，0.5％多聚蛋白胨，0.3％酵母抽提物，0.6％麥芽，0.2％脫脂大豆餅，0.2％CaCO3，0.02％消泡劑(Silicon KM-72，由Shin-Etsu Chemical制造)滅菌之前pH7.0，2個6mm直徑的小珠)中并在振蕩器上28℃培養(yǎng)22小時，并用作種子培養(yǎng)物。其后，將1.0的種子培養(yǎng)物接種在250ml容量錐形燒瓶中所含的30ml的生產(chǎn)培養(yǎng)基(1％葡萄糖，1％蛋白胨，0.5％肉汁，0.4％植物蛋白胨，3％大豆原油，0.2％NaCl，0.3％CaCO3，0.08％乳化劑(Nikkol CO-20TX，由Nikko Chemical制造)，pH 7.0)中并在振蕩器上28℃培養(yǎng)67到77小時。用50％硫酸將培養(yǎng)液體調(diào)節(jié)到pH4.0或者更少然后過濾，并將其用作分析樣品。對于分析法，在以下條件下進行檢測柱；YMC-PackODS-AM(S-5μm，6.0×150mm，YMC)，流動相；緩沖劑(0.01MCH3COONH4，0.0001M K2HPO4，pH 6.05)∶CH3CN∶C2H5OH＝3∶3∶2，柱溫；35℃，流速；1.20ml/min，檢測波長232nm和280nm。
結(jié)果，在菌株938-15的情況下總麥迪霉素量為798μg/ml，在菌株1149-38(FERM BP-10501)情況下為1127μg/ml。
本申請是基于2005年3月31日提交的日本專利申請JP2005-101836，其全部內(nèi)容在此處引入作為參考，無異于詳細的陳述。
序列表<110>明治制菓株式會社(Meiji Seika Kaihsa，Ltd)<120>麥迪霉素高產(chǎn)菌株(Midecamycin hyper producing strain)<130>SPI060897-66<150>P2005-101836<151>2005-03-31<160>11<170>PatentIn version 3.1<210>1<211>19<212>DNA<213>Artificial<220>
<223>Synthetic primer<400>1cggtsaagtc saacatcgg 19<210>2<211>20<212>DNA<213>Artificial<220>
<223>Synthetic primer<400>2gcratctcrc cctgcgartg 20<210>3<211>13536<212>DNA<213>Streptomyces mycarofaciens<220>
<221>CDS<222>(1)..(13533)<223>
<400>3atg ctg gtg tct gga gat ctc gtg act tcc cga att gac gac cga tcc48
Met Leu Val Ser Gly Asp Leu Val Thr Ser Arg Ile Asp Asp Arg Ser1 5 10 15gat gca att gcc gtt gtc gga atg tcc tgt cga ttt ccc ggc gcc ccg96Asp Ala Ile Ala Val Val Gly Met Ser Cys Arg Phe Pro Gly Ala Pro20 25 30gga gtc gaa gaa ttc tgg aaa ctg ctg acc gac gga acg gaa gcc gtc144Gly Val Glu Glu Phe Trp Lys Leu Leu Thr Asp Gly Thr Glu Ala Val35 40 45agt cgc gcg gcc gat ggc cgt cgg cgc ggc atg atc gag gcg gtc ggc192Ser Arg Ala Ala Asp Gly Arg Arg Arg Gly Met Ile Glu Ala Val Gly50 55 60gac ttc gac gcc acg ttc ttc ggc atg tca ccg cgc gag gcc gcc gag240Asp Phe Asp Ala Thr Phe Phe Gly Met Ser Pro Arg Glu Ala Ala Glu65 70 75 80acc gat ccg cag cag cgc ctg ctg ctc gaa ctc ggc tgg gag gcc ctg288Thr Asp Pro Gln Gln Arg Leu Leu Leu Glu Leu Gly Trp Glu Ala Leu85 90 95gag gac gcc gga atc gtc ccg ggg tcg ctg cgc ggc gag gcg gtc ggc336Glu Asp Ala Gly Ile Val Pro Gly Ser Leu Arg Gly Glu Ala Val Gly100 105 110atc ttc gtc ggt gcc atg cac aac gac tac gcc acc ctg ctg cac cgg384Ile Phe Val Gly Ala Met His Asn Asp Tyr Ala Thr Leu Leu His Arg115 120 125gcc ggc gca ccg gcc ggc gcc cac acc gcc acc ggc ctc cag ccc gcc432Ala Gly Ala Pro Ala Gly Ala His Thr Ala Thr Gly Leu Gln Pro Ala130 135 140atg ctc gcc aac cgg ctc tcc tac gtc ctg gga acg cgc ggc ccc agc480Met Leu Ala Asn Arg Leu Ser Tyr Val Leu Gly Thr Arg Gly Pro Ser145 150 155 160ctg gcg gtt gac acc gcg cag tcg tcg tcg ctg gtc gcc gtg gcc ctc528Leu Ala Val Asp Thr Ala Gln Ser Ser Ser Leu Val Ala Val Ala Leu165 170 175gcg gtc gag agc ctg cgc gcc gga acc tcc cgc atc gcc atc gca ggc576Ala Val Glu Ser Leu Arg Ala Gly Thr Ser Arg Ile Ala Ile Ala Gly180 185 190ggc gtc aac ctg atc ctc gcc gac gag ggc tcg gcc acc atg gag cgg624Gly Val Asn Leu Ile Leu Ala Asp Glu Gly Ser Ala Thr Met Glu Arg195 200 205
ctc ggc gcg ctc tcc ccc gac ggg cgt tgc tac acc ttc gac gcc cgc672Leu Gly Ala Leu Ser Pro Asp Gly Arg Cys Tyr Thr Phe Asp Ala Arg210 215 220gcc aac ggc tat gtg cgt ggc gag ggc ggt gcc gcc gtc gta ctg aag720Ala Asn Gly Tyr Val Arg Gly Glu Gly Gly Ala Ala Val Val Leu Lys225 230 235 240ccc ctc gcc gac gcc ttg gcc gac ggc gac ccg gtg tac tgc gtg gtg768Pro Leu Ala Asp Ala Leu Ala Asp Gly Asp Pro Val Tyr Cys Val Val245 250 255cgc agc gcc gcc act ggc aac gac ggc ggc ggc ccc ggg ctg acc tcc816Arg Ser Ala Ala Thr Gly Asn Asp Gly Gly Gly Pro Gly Leu Thr Ser260 265 270ccc gac cac gaa ggc cag gaa gcc gtg ctc cgg gcg gcc tgc gcc cag864Pro Asp His Glu Gly Gln Glu Ala Val Leu Arg Ala Ala Cys Ala Gln275 280 285gcc gga gtc gac ccc gca aag gtg cgc ttc gtc gaa ctg cac ggc acc912Ala Gly Val Asp Pro Ala Lys Val Arg Phe Val Glu Leu His Gly Thr290 295 300ggc acc ccc gtg ggc gac ccg gtc gag gca cgg gcc ctg ggt gcg gtc960Gly Thr Pro Val Gly Asp Pro Val Glu Ala Arg Ala Leu Gly Ala Val305 310 315 320cac ggc tcc ggg cgg ccg gcg gac gca ccc ctg ctg gtg ggc tcc gtg1008His Gly Ser Gly Arg Pro Ala Asp Ala Pro Leu Leu Val Gly Ser Val325 330 335aag acc aac atc ggc cac ctg gaa ggc gca gcc ggc atc gcg ggg ctg1056Lys Thr Asn Ile Gly His Leu Glu Gly Ala Ala Gly Ile Ala Gly Leu340 345 350gtc aag gcc gca ctc tgc ctg cgg aat cgc acc ctg ccc ggc tcg ctc1104Val Lys Ala Ala Leu Cys Leu Arg Asn Arg Thr Leu Pro Gly Ser Leu355 360 365aac ttc gtc acc ccc cac ccc gcc atc cct ctg gac cgg ctc cgg ctg1152Asn Phe Val Thr Pro His Pro Ala Ile Pro Leu Asp Arg Leu Arg Leu370 375 380aag gtg cag acg acc ccg acc acg ctg cac ccc gat ccg gac ggc tcc1200Lys Val Gln Thr Thr Pro Thr Thr Leu His Pro Asp Pro Asp Gly Ser385 390 395 400ccc ctg ctg gcg ggt gtc agc tcc ttc ggt atc ggc ggc acc aac tgc1248Pro Leu Leu Ala Gly Val Ser Ser Phe Gly Ile Gly Gly Thr Asn Cys405 410 415
cat gtc gtc ctg gag cac ctg ccc gag ccg gcc ccc acc aca agg gaa1296His Val Val Leu Glu His Leu Pro Glu Pro Ala Pro Thr Thr Arg Glu420 425 430gcc cta ccc gcc ccg cac ctg gtc ccg ccc ctg ctg ttg tcg gcc cgt1344Ala Leu Pro Ala Pro His Leu Val Pro Pro Leu Leu Leu Ser Ala Arg435 440 445tcc cac ccg gca ctg ctg gcc cag gcg gcg cgg ctc cgt gac cac ctg1392Ser His Pro Ala Leu Leu Ala Gln Ala Ala Arg Leu Arg Asp His Leu450 455 460agc cgc acc gct gcc gac ccg cag gac gtc gct tac tcc ctg gcc acc1440Ser Arg Thr Ala Ala Asp Pro Gln Asp Val Ala Tyr Ser Leu Ala Thr465 470 475 480aca cgc tcc ctc ttc gag cac cgc gcc gcg ctg ccc tgc ggc aac cgc1488Thr Arg Ser Leu Phe Glu His Arg Ala Ala Leu Pro Cys Gly Asn Arg485 490 495gag gag ttg gtc gcc gcc ctc gac gca ctc gcc cac ggc agg atc acg1536Glu Glu Leu Val Ala Ala Leu Asp Ala Leu Ala His Gly Arg Ile Thr500 505 510gcg ggc gtg cga gtc gac tcg gct gtg tcg ggt ggg acg gct gtg ttg1584Ala Gly Val Arg Val Asp Ser Ala Val Ser Gly Gly Thr Ala Val Leu515 520 525ttt acg ggt cag ggt gcg cag tgg gtt ggt atg ggg cgt gag ttg tat1632Phe Thr Gly Gln Gly Ala Gln Trp Val Gly Met Gly Arg Glu Leu Tyr530 535 540ggg ttg gat ggg gtg ttt gct gcg gcg ttg gat gag gtt ttg ggt gtg1680Gly Leu Asp Gly Val Phe Ala Ala Ala Leu Asp Glu Val Leu Gly Val545 550 555 560gtg ggg gag gtg ggt ggt tgg tct ttg cgt gag gtg atg ttt ggt gag1728Val Gly Glu Val Gly Gly Trp Ser Leu Arg Glu Val Met Phe Gly Glu565 570 575ggt ggt ggt gtt ggg gtg ggg ttg ttg gat ggt acg gag ttt gcg cag1776Gly Gly Gly Val Gly Val Gly Leu Leu Asp Gly Thr Glu Phe Ala Gln580 585 590cct gct ttg ttt gcg ttg gag gtg gcg ttg ttt cgg gct gtg gag gct1824Pro Ala Leu Phe Ala Leu Glu Val Ala Leu Phe Arg Ala Val Glu Ala595 600 605cgg ggg gtg cgg gct tcg gtg gtg ttg ggg cat tcg gtg ggg gag gtt1872Arg Gly Val Arg Ala Ser Val Val Leu Gly His Ser Val Gly Glu Val
610 615 620gct gct gcg tgt gtg gcg ggg gtg ttt tcg ctt gcg gat gcg gcg cgg1920Ala Ala Ala Cys Val Ala Gly Val Phe Ser Leu Ala Asp Ala Ala Arg625 630 635 640ttg gtg gtg gcg cgt ggt cgg ttg atg ggt gcg ttg cct gtg ggt ggg1968Leu Val Val Ala Arg Gly Arg Leu Met Gly Ala Leu Pro Val Gly Gly645 650 655ggg atg ttg tcg gtt cgt gcg tct gag gcc gaa ctt gtt gat gtt gtg2016Gly Met Leu Ser Val Arg Ala Ser Glu Ala Glu Leu Val Asp Val Val660 665 670gct ggg ttg ggt ggt cgg gtg tcg gtg gct gcg gtc aat ggt ccg gcg2064Ala Gly Leu Gly Gly Arg Val Ser Val Ala Ala Val Asn Gly Pro Ala675 680 685tcg gtg gtg ttg tct ggt gag tgt ggt gcg ttg gat gtt gtt gcg gcg2112Ser Val Val Leu Ser Gly Glu Cys Gly Ala Leu Asp Val Val Ala Ala690 695 700cgg ttg ggt ggg cgt ggg gtg gag tgc aag cgg ttg gtg gtg tcg cat2160Arg Leu Gly Gly Arg Gly Val Glu Cys Lys Arg Leu Val Val Ser His705 710 715 720gcg ttt cat tcg gcg ttg atg gat ccg atg ttg gag gag ttt cgt ggg2208Ala Phe His Ser Ala Leu Met Asp Pro Met Leu Glu Glu Phe Arg Gly725 730 735gtt gct gag agt gtg gag tat cgg cgg ccg tgt gtg ccg gtg gtg tcg2256Val Ala Glu Ser Val Glu Tyr Arg Arg Pro Cys Val Pro Val Val Ser740 745 750aat gtg acg ggt ggg gtg gtt ggg ttt gat gag ttg ggt tgt gcc gag2304Asn Val Thr Gly Gly Val Val Gly Phe Asp Glu Leu Gly Cys Ala Glu755 760 765tat tgg gtg cgg cat gcg cgg gag gcg gtg cgt ttc gct gag ggg att2352Tyr Trp Val Arg His Ala Arg Glu Ala Val Arg Phe Ala Glu Gly Ile770 775 780cgg gct gct cgt gct ctt ggt gtg gat acg ttc ctg gag gtg ggt ccg2400Arg Ala Ala Arg Ala Leu Gly Val Asp Thr Phe Leu Glu Val Gly Pro785 790 795 800cat gcg gtt ttg acg gcg atg gct ggt cag tgt ctt gat gct gag gag2448His Ala Val Leu Thr Ala Met Ala Gly Gln Cys Leu Asp Ala Glu Glu805 810 815gct gac ttg gcg ttt gtg ccg gtc ctg cgg cgt gat cgg ccg gca ttg2496
Ala Asp Leu Ala Phe Val Pro Val Leu Arg Arg Asp Arg Pro Ala Leu820 825 830cag acc ttc acc acc gca ctc gcc act ctg cac acc cgt gat gcc gaa2544Gln Thr Phe Thr Thr Ala Leu Ala Thr Leu His Thr Arg Asp Ala Glu835 840 845ctc gac gcc gtg gcg ctc cat tca ggc agc gat gcc cgg cgg atc gac2592Leu Asp Ala Val Ala Leu His Ser Gly Ser Asp Ala Arg Arg Ile Asp850 855 860ctg ccc acc tac ccc ttc caa cgc cgt act cac tgg tcg ccg gcg ctg2640Leu Pro Thr Tyr Pro Phe Gln Arg Arg Thr His Trp Ser Pro Ala Leu865 870 875 880agc cac gga cac gcg gcc ggc gtc gtg cgg gcc tcg acc gct acc gag2688Ser His Gly His Ala Ala Gly Val Val Arg Ala Ser Thr Ala Thr Glu885 890 895atc cgg ggg aac gac gag atc ccg gag agt gcc gag gca ctc ctt cgg2736Ile Arg Gly Asn Asp Glu Ile Pro Glu Ser Ala Glu Ala Leu Leu Arg900 905 910gac ccg gcc gac ggg tcg ctc gcg gca tcc ccg gag ccg gcg aca ccc2784Asp Pro Ala Asp Gly Ser Leu Ala Ala Ser Pro Glu Pro Ala Thr Pro915 920 925gac cag ctc gtc cgg ctg gtc cgc gag acc act gct gcc gtc ctg ggc2832Asp Gln Leu Val Arg Leu Val Arg Glu Thr Thr Ala Ala Val Leu Gly930 935 940cac gac gac ccc gac gag atc gtc ctc gac cgc acc ttc acc tct cag2880His Asp Asp Pro Asp Glu Ile Val Leu Asp Arg Thr Phe Thr Ser Gln945 950 955 960ggc ctg gaa tcg gtg acc gcg gtc gaa ctc cgc gac cta ctg aac cgg2928Gly Leu Glu Ser Val Thr Ala Val Glu Leu Arg Asp Leu Leu Asn Arg965 970 975gcc acg ggg ctg acc ctc gcg gcc acg ctc gtc tac gac ctg ccc acc2976Ala Thr Gly Leu Thr Leu Ala Ala Thr Leu Val Tyr Asp Leu Pro Thr980 985 990ccg cgc gcc gtc gcc gat tac ctg tcg gcc gcg atg ctc gcg acc gac 3024Pro Arg Ala Val Ala Asp Tyr Leu Ser Ala Ala Met Leu Ala Thr Asp995 1000 1005gat gcg aac tcc agc gcg cac caa acc acc gcg gcg gcg acc acc 3069Asp Ala Asn Ser Ser Ala His Gln Thr Thr Ala Ala Ala Thr Thr1010 1015 1020
cgg agc ggt gcg cgg aac gac gac ccg atc gcc atc gtc ggc gtc3114Arg Ser Gly Ala Arg Asn Asp Asp Pro Ile Ala Ile Val Gly Val1025 1030 1035ggc tcc cac ttc ccc ggc ggc gtg gac tcg cgc gcc ggc ctg tgg3159Gly Ser His Phe Pro Gly Gly Val Asp Ser Arg Ala Gly Leu Trp1040 1045 1050gat ctg ctg gcc tcc ggc acc gac gcg atc tcg tcc ttt ccc acc3204Asp Leu Leu Ala Ser Gly Thr Asp Ala Ile Ser Ser Phe Pro Thr1055 1060 1065gac cgt ggt tgg gat ctc aac gag ctg tac gac ccc gag ccc ggc3249Asp Arg Gly Trp Asp Leu Asn Glu Leu Tyr Asp Pro Glu Pro Gly1070 1075 1080atc ccc ggc aag acc tat gtg cgt cag ggc ggc ttc ctg cat cag3294Ile Pro Gly Lys Thr Tyr Val Arg Gln Gly Gly Phe Leu His Gln1085 1090 1095gcg gcc gag ttc gac gcg gag ttc ttc ggc atc tcg ccg cgc gag3339Ala Ala Glu Phe Asp Ala Glu Phe Phe Gly Ile Ser Pro Arg Glu1100 1105 1110gcg acc gcc atg gac ccc cag cag cgg ctg ctg ctg gag acc tcc3384Ala Thr Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Thr Ser1115 1120 1125tgg gag gcg ctg gag gac gcc gga gtg tgc ccc gag tcg ctg cgc3429Trp Glu Ala Leu Glu Asp Ala Gly Val Cys Pro Glu Ser Leu Arg1130 1135 1140ggc acc aac acc ggc gtg ttc atc ggc gca gtc gca ccg gag tac3474Gly Thr Asn Thr Gly Val Phe Ile Gly Ala Val Ala Pro Glu Tyr1145 1150 1155ggc ccg agg ctc cac gag gga gcg gac ggg tac gag ggg tat ctg3519Gly Pro Arg Leu His Glu Gly Ala Asp Gly Tyr Glu Gly Tyr Leu1160 1165 1170ctc acc ggc acc acg gcg agc gtg gcc tcc ggc cgg atc gcc tac3564Leu Thr Gly Thr Thr Ala Ser Val Ala Ser Gly Arg Ile Ala Tyr1175 1180 1185acc ttc ggc acg cgc ggg ccg gcg ctc acg gtg gat acc gcg tgt3609Thr Phe Gly Thr Arg Gly Pro Ala Leu Thr Val Asp Thr Ala Cys1190 1195 1200tcg tcg tcg ttg gtg gcg ttg cac ctg gcg gtg cag tcg ttg cgg3654Ser Ser Ser Leu Val Ala Leu His Leu Ala Val Gln Ser Leu Arg1205 1210 1215
cgg ggt gag tgt gat atg gcg ttg gcc ggc gga gcc acg gtg atg3699Arg Gly Glu Cys Asp Met Ala Leu Ala Gly Gly Ala Thr Val Met1220 1225 1230tcc ggc ccc ggc atg ttc gtg gag ttc tcc cgg cag cgt ggg ttg3744Ser Gly Pro Gly Met Phe Val Glu Phe Ser Arg Gln Arg Gly Leu1235 1240 1245gcg tcg gat ggg cgg tgc aag gcg ttc tcc gcc gat gcc gac ggc3789Ala Ser Asp Gly Arg Cys Lys Ala Phe Ser Ala Asp Ala Asp Gly1250 1255 1260acg gcc tgg tcc gag ggc gtc gcc gtt ctg gcg ctg gag cgt ctc3834Thr Ala Trp Ser Glu Gly Val Ala Val Leu Ala Leu Glu Arg Leu1265 1270 1275tcc gac gcc cgc cgc gcc ggt cac cgg gtg ctg gcg ctg gtc cgg3879Ser Asp Ala Arg Arg Ala Gly His Arg Val Leu Ala Leu Val Arg1280 1285 1290ggc agc gcg gtc aac cag gac ggc gcc agc aac ggt ctc acc gcg3924Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala1295 1300 1305ccc agc ggt ccc gcg cag gag agt gtc atc cgt gag gcg ttg gcg3969Pro Ser Gly Pro Ala Gln Glu Ser Val Ile Arg Glu Ala Leu Ala1310 1315 1320gat gcc ggg ttg ggg ccg ggt gat gtg gat gtg gtg gag gcg cat4014Asp Ala Gly Leu Gly Pro Gly Asp Val Asp Val Val Glu Ala His1325 1330 1335ggt acg ggt acg gcg ttg ggt gat ccg atc gag gct ggt gcg ttg4059Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile Glu Ala Gly Ala Leu1340 1345 1350ctg gcc acg tat gga tgt gag cgg gtg ggt gat ccg ttg tgg ttg4104Leu Ala Thr Tyr Gly Cys Glu Arg Val Gly Asp Pro Leu Trp Leu1355 1360 1365ggg tcg ctg aag tcc aac atc ggg cac act cag gcc gcc gcg ggt4149Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gln Ala Ala Ala Gly1370 1375 1380gtc gcc ggt gtc atc aag atg gtg gag gcc ctg cgc cat ggc acg4194Val Ala Gly Val Ile Lys Met Val Glu Ala Leu Arg His Gly Thr1385 1390 1395ctg ccg cgg acg ctc cac gcc gac cgc ccc agc aca cac gtc gac4239Leu Pro Arg Thr Leu His Ala Asp Arg Pro Ser Thr His Val Asp
1400 1405 1410tgg tct tcg ggg ggc gtg gag ttg ctg acc gag gcg cgc ccg tgg4284Trp Ser Ser Gly Gly Val Glu Leu Leu Thr Glu Ala Arg Pro Trp1415 1420 1425ccg gag cgg gag ggc cgg ccg cgg cgg gcc gcg gtg tcg gcc ttc4329Pro Glu Arg Glu Gly Arg Pro Arg Arg Ala Ala Val Ser Ala Phe1430 1435 1440ggt gtc agc ggt acc aac gct cac ctg gtc att gaa gag ccc ccc4374Gly Val Ser Gly Thr Asn Ala His Leu Val Ile Glu Glu Pro Pro1445 1450 1455gtg gag ttg cct gct ggt gct ggt gct ggt gct ggt gct ggt gct4419Val Glu Leu Pro Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Ala1460 1465 1470ggg gtg tct tcg gtt gtg gcg tgg ccg ttg tcg gct cgt tcg ggt4464Gly Val Ser Ser Val Val Ala Trp Pro Leu Ser Ala Arg Ser Gly1475 1480 1485gag gcg ttg cgg gcg cag gcg gtg cgg ttg cgt gag cat gtg gag4509Glu Ala Leu Arg Ala Gln Ala Val Arg Leu Arg Glu His Val Glu1490 1495 1500cgt gtt ggg gct gat ccg gtt gat gtt gcc ttt tcg ttg gcg gtg4554Arg Val Gly Ala Asp Pro Val Asp Val Ala Phe Ser Leu Ala Val1505 1510 1515acg cgt gcg tcg ttc ggt gag cgt gcg gtg gtc gtt ggt ggt gac4599Thr Arg Ala Ser Phe Gly Glu Arg Ala Val Val Val Gly Gly Asp1520 1525 1530cgt gcg gag ttg ctg gcg ggg ctt gat gcg ctt gct ggg ggg cgt4644Arg Ala Glu Leu Leu Ala Gly Leu Asp Ala Leu Ala Gly Gly Arg1535 1540 1545cgg ggg ccg ggg gtt gtc cgg ggc tcg gct gtg tcg ggt ggg acg4689Arg Gly Pro Gly Val Val Arg Gly Ser Ala Val Ser G1y Gly Thr1550 1555 1560gct gtg ttg ttt acg ggt cag ggt gcg cag tgg gtt ggt atg ggg4734Ala Val Leu Phe Thr Gly Gln Gly Ala Gln Trp Val Gly Met Gly1565 1570 1575cgt gag ttg tat ggg ttg gat ggg gtg ttt gct gcg gcg ttg gat4779Arg Glu Leu Tyr Gly Leu Asp Gly Val Phe Ala Ala Ala Leu Asp1580 1585 1590gag gtg ttg ggt gtg gtg ggg gag gtg ggt ggt tgg tct ttg cgt4824
Glu Val Leu Gly Val Val Gly Glu Val Gly Gly Trp Ser Leu Arg1595 1600 1605gag gtg atg ttt ggt gag ggt ggt ggt gtt ggg gtg ggg ttg ttg4869Glu Val Met Phe Gly Glu Gly Gly Gly Val Gly Val Gly Leu Leu1610 1615 1620gat ggt acg gag ttt gcg cag cct gct ttg ttt gcg ttg gag gtg4914Asp Gly Thr Glu Phe Ala Gln Pro Ala Leu Phe Ala Leu Glu Val1625 1630 1635gcg ttg ttt cgg gct gtg gag gct cgg ggg gtg cgg gct tcg gtg4959Ala Leu Phe Arg Ala Val Glu Ala Arg Gly Val Arg Ala Ser Val1640 1645 1650gtg ttg ggg cat tcg gtg ggg gag gtt gct gct gcg tgt gtg gcg5004Val Leu Gly His Ser Val Gly Glu Val Ala Ala Ala Cys Val Ala1655 1660 1665ggg gtg ttt tcg ctt gcg gat gcg gcg cgg ttg gtg gtg gcg cgt5049Gly Val Phe Ser Leu Ala Asp Ala Ala Arg Leu Val Val Ala Arg1670 1675 1680ggt cgg ttg atg ggt ggg ttg cct gtg ggt ggg ggg atg ttg tcg5094Gly Arg Leu Met Gly Gly Leu Pro Val Gly Gly Gly Met Leu Ser1685 1690 1695gtt cgt gcg tct gag gcc gaa ctt gct gat gtt gtg gct ggg ttg5139Val Arg Ala Ser Glu Ala Glu Leu Ala Asp Val Val Ala Gly Leu1700 1705 1710ggt ggt cgg gtg tcg gtg gct gcg gtc aat ggt ccg gcg tcg gtg5184Gly Gly Arg Val Ser Val Ala Ala Val Asn Gly Pro Ala Ser Val1715 1720 1725gtg ttg tct ggt gag tgt ggt gcg ttg gat gtt gtt gcg gcg cgg5229Val Leu Ser Gly Glu Cys Gly Ala Leu Asp Val Val Ala Ala Arg1730 1735 1740ttg ggt ggg cgt ggg gtg gag tgc aag cgg ttg gtg gtg tcg cat5274Leu Gly Gly Arg Gly Val Glu Cys Lys Arg Leu Val Val Ser His1745 1750 1755gcg ttt cat tcg gcg ttg atg gag ccg atg ttg gag gag ttt cgt5319Ala Phe His Ser Ala Leu Met Glu Pro Met Leu Glu Glu Phe Arg1760 1765 1770ggg gtt gct gag agt gtg gag tat cgg cgg ccg tgt gtg ccg gtg5364Gly Val Ala Glu Ser Val Glu Tyr Arg Arg Pro Cys Val Pro Val1775 1780 1785
gtg tcg aat gtg acg ggt ggg gtg gtt ggg ttt gat gag ttg ggt5409Val Ser Asn Val Thr Gly Gly Val Val Gly Phe Asp Glu Leu Gly1790 1795 1800tgt gcc gag tat tgg gtg cgg cat gcg cgg gag gcg gtg cgt ttc5454Cys Ala Glu Tyr Trp Val Arg His Ala Arg Glu Ala Val Arg Phe1805 1810 1815gct gag ggg ata cgg gct gct cgt gct ctt ggt gtg gat acg ttc5499Ala Glu Gly Ile Arg Ala Ala Arg Ala Leu Gly Val Asp Thr Phe1820 1825 1830ctg gag gtt ggt ccg cat gcg gtt ttg acg gcg atg gct ggt cag5544Leu Glu Val Gly Pro His Ala Val Leu Thr Ala Met Ala Gly Gln1835 1840 1845tgt ctt gat gga gag gag gct gac ttg gcg ttt gtg ccg gtc ctg5589Cys Leu Asp Gly Glu Glu Ala Asp Leu Ala Phe Val Pro Val Leu1850 1855 1860cgg cgt gat cgg ccg gca tcg cag acc ttc acc acc gca ctc gcc5634Arg Arg Asp Arg Pro Ala Ser Gln Thr Phe Thr Thr Ala Leu Ala1865 1870 1875acg ctt tgt gtt cgg ggc act gag gtc gat tgg gcc acg ccg cac5679Thr Leu Cys Val Arg Gly Thr Glu Val Asp Trp Ala Thr Pro His1880 1885 1890cgg aag agt ggt gca caa cgc att gac ctg ccc acg tac ccc ttc5724Arg Lys Ser Gly Ala Gln Arg Ile Asp Leu Pro Thr Tyr Pro Phe1895 1900 1905cag cgc gcc cga tac tgg ctt gac ccc gcc cct gca atg gcg ctc5769Gln Arg Ala Arg Tyr Trp Leu Asp Pro Ala Pro Ala Met Ala Leu1910 1915 1920act acc gtg gcc gcc agt tcg gcc gag gcc gcg gcg acg gcc act5814Thr Thr Val Ala Ala Ser Ser Ala Glu Ala Ala Ala Thr Ala Thr1925 1930 1935gag ggg aca gcc ctg gaa acg gcc ggg ctc cgc tac cgc atc gcc5859Glu Gly Thr Ala Leu Glu Thr Ala Gly Leu Arg Tyr Arg Ile Ala1940 1945 1950tgg cag gcc gcc gcc acg gac cgc ggc acc tct cgc tcg gcg ggg5904Trp Gln Ala Ala Ala Thr Asp Arg Gly Thr Ser Arg Ser Ala Gly1955 1960 1965cac gtg gtg cta ctc acc tcg gac gac gac gcg acc gaa tcc gga5949His Val Val Leu Leu Thr Ser Asp Asp Asp Ala Thr Glu Ser Gly1970 1975 1980
ctt gcc gcc gcg att acc cgc gaa ctc gcc gtg cgc ggc gcc gag5994Leu Ala Ala Ala Ile Thr Arg Glu Leu Ala Val Arg Gly Ala Glu1985 1990 1995gta cgc acc gcg atc ctg cca gtc ggc acc gac cgc gag acg gcc6039Val Arg Thr Ala Ile Leu Pro Val Gly Thr Asp Arg Glu Thr Ala2000 2005 2010gca gac ctg cta cga acc tcc ggt gac ggc gcc gca cgc agc acg6084Ala Asp Leu Leu Arg Thr Ser Gly Asp Gly Ala Ala Arg Ser Thr2015 2020 2025cac gtc ctg tgg ctc gcc ccg gcc gag ccc gac acg gcc gac gcc6129His Val Leu Trp Leu Ala Pro Ala Glu Pro Asp Thr Ala Asp Ala2030 2035 2040gtc gcg ctg atc cag gcc ctg ggc gag gca ggg cac gac gcc cca6174Val Ala Leu Ile Gln Ala Leu Gly Glu Ala Gly His Asp Ala Pro2045 2050 2055ctg tgg atc gcc acg cgt gac gcg gtg gcc gtc cag ccg ggc gag6219Leu Trp Ile Ala Thr Arg Asp Ala Val Ala Val Gln Pro Gly Glu2060 2065 2070aag ctg tcc gtc gcc gga gcg cag ctc tgg ggg ctc ggg cag gtc6264Lys Leu Ser Val Ala Gly Ala Gln Leu Trp Gly Leu Gly Gln Val2075 2080 2085gcc gcc ctc gaa ctg ttc cag cgc tgg ggc ggc ctg gtg gac ctg6309Ala Ala Leu Glu Leu Phe Gln Arg Trp Gly Gly Leu Val Asp Leu2090 2095 2100ccc gag aac ccg tcg ccc gct gcg gtc cgc gcg ttc gtc ggg gcg6354Pro Glu Asn Pro Ser Pro Ala Ala Val Arg Ala Phe Val Gly Ala2105 2110 2115ctg ttc gcg gag ggt gac gac aac cag atc gcg gtg cgg ccc tcc6399Leu Phe Ala Glu Gly Asp Asp Asn Gln Ile Ala Val Arg Pro Ser2120 2125 2130ggc gtg tac gtc cgc cgc gtg gcc ccc gcc ccc gcc ccc gct ccc6444Gly Val Tyr Val Arg Arg Val Ala Pro Ala Pro Ala Pro Ala Pro2135 2140 2145gcc ctc atc ggg cag gct gcg cag gac gac cgg tcc ggc ccg tcc6489Ala Leu Ile Gly Gln Ala Ala Gln Asp Asp Arg Ser Gly Pro Ser2150 2155 2160gat gga ctc gat ggg aac aat gga acc gcg ccg gtg aac tgg cac6534Asp Gly Leu Asp Gly Asn Asn Gly Thr Ala Pro Val Asn Trp His
2165 2170 2175ccc tcc ggc acc gta ctg atc acc ggt ggc acc ggg gcc ctc ggc6579Pro Ser Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ala Leu Gly2180 2185 2190gca cag gtg gcc cgc agg ctc gcc cga gcg ggc gcg ccg cat ctg6624Ala Gln Val Ala Arg Arg Leu Ala Arg Ala Gly Ala Pro His Leu2195 2200 2205ctc ctg gtc agc cgc cgt gga ccg gac ggc cct ggt acg ggc gaa6669Leu Leu Val Ser Arg Arg Gly Pro Asp Gly Pro Gly Thr Gly Glu2210 2215 2220ctg gtc ggg gaa ctg aca gcg cac ggc acc gaa gtg acc gtc acg6714Leu Val Gly Glu Leu Thr Ala His Gly Thr Glu Val Thr Val Thr2225 2230 2235gcc tgt gac gcc gcc gac cgc gat gcg ctc gcc gag ctg ctc gcg6759Ala Cys Asp Ala Ala Asp Arg Asp Ala Leu Ala Glu Leu Leu Ala2240 2245 2250agc att ccc gag gat cgc ccc ctc acc gcc gta ctg cac gcg gca6804Ser Ile Pro Glu Asp Arg Pro Leu Thr Ala Val Leu His Ala Ala2255 2260 2265ggt gtg ctc gac gac ggc gtg ctc gac gcg ctc acc ccc gat cgg6849Gly Val Leu Asp Asp Gly Val Leu Asp Ala Leu Thr Pro Asp Arg2270 2275 2280ctc gac gcc gta ctg cgc gcc aag gta acc gtg gcc cgc cac ctg6894Leu Asp Ala Val Leu Arg Ala Lys Val Thr Val Ala Arg His Leu2285 2290 2295gac gag ctc acc gca ggc ata ccg ctg gat gcc ttt gtg ctc ttc6939Asp Glu Leu Thr Ala Gly Ile Pro Leu Asp Ala Phe Val Leu Phe2300 2305 2310tcc tcc atc gtc ggg gtg tgg ggc aac ggc ggc cag ggc ggc tat6984Ser Ser Ile Val Gly Val Trp Gly Asn Gly Gly Gln Gly Gly Tyr2315 2320 2325gcg gcg gcc aac gcc gcg ctc gat gcc ctg gcg cac cgg cgc cgg7029Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu Ala His Arg Arg Arg2330 2335 2340gcc cgg gga cag cgt gcc acg tcg att gcc tgg ggg ccg tgg gcc7074Ala Arg Gly Gln Arg Ala Thr Ser Ile Ala Trp Gly Pro Trp Ala2345 2350 2355ggc gcc gga atg gcg gcc ggc gca ggc tcg aag gcc ttc cag cgg7119
Gly Ala Gly Met Ala Ala Gly Ala Gly Ser Lys Ala Phe Gln Arg2360 2365 2370gat ggc atc cag gct ctg gat ccc gag cgt gca ctc aat gtg ctg7164Asp Gly Ile Gln Ala Leu Asp Pro Glu Arg Ala Leu Asn Val Leu2375 2380 2385gac gac gtg gtt cgc gcc gac gag acg tct gtg gcc gcc gag ccc7209Asp Asp Val Val Arg Ala Asp Glu Thr Ser Val Ala Ala Glu Pro2390 2395 2400tct ttg atc gtc gcc gat gtg gac tgg agc acg ttc gtc ggg cgc7254Ser Leu Ile Val Ala Asp Val Asp Trp Ser Thr Phe Val Gly Arg2405 24l0 2415tcc gtc gcc cga cgc acc tgg gcg ctt ttc gac ggt gtt ccg gcc7299Ser Val Ala Arg Arg Thr Trp Ala Leu Phe Asp Gly Val Pro Ala2420 2425 2430gcc tgc tcc gcg cgt tcc gcc cag gcc gca cag ggc cgt tcc gcg7344Ala Cys Ser Ala Arg Ser Ala Gln Ala Ala Gln Gly Arg Ser Ala2435 2440 2445cac gcc ccg gga gag cgg ccg cac cac ggc ggc att ggt ggg agc7389His Ala Pro Gly Glu Arg Pro His His Gly Gly Ile Gly Gly Ser2450 2455 2460gga gac gga gcg gac gag gac cgc ccc tgg ctc tct gcc ggc ccc7434Gly Asp Gly Ala Asp Glu Asp Arg Pro Trp Leu Ser Ala Gly Pro2465 2470 2475tcc tcg ccg gaa cgg cgg cgg gca ctg ctc gac ttg gtg cgc tcc7479Ser Ser Pro Glu Arg Arg Arg Ala Leu Leu Asp Leu Val Arg Ser2480 2485 2490gag gcc gcc gag atc ctg cgt cac ggt tcg gct gcc gcg gtc gac7524Glu Ala Ala Glu Ile Leu Arg His Gly Ser Ala Ala Ala Val Asp2495 2500 2505ccg gag acc gcg ttc cgg gcc gcc ggg ttc gac tcc ctc acc gtg7569Pro Glu Thr Ala Phe Arg Ala Ala Gly Phe Asp Ser Leu Thr Val2510 2515 2520ctc gaa ctg cgt aat cgt ctg acc gcc gcc atc ggg ctg aac ctg7614Leu Glu Leu Arg Asn Arg Leu Thr Ala Ala Ile Gly Leu Asn Leu2525 2530 2535ccg agc acc ctg ctg ttc gac tat ccg aac ccg aac gcc ctg gcc7659Pro Ser Thr Leu Leu Phe Asp Tyr Pro Asn Pro Asn Ala Leu Ala2540 2545 2550
gac cat ctg cac gac gaa ttg ttc ggt gct gac agc gaa gca ccg7704Asp His Leu His Asp Glu Leu Phe Gly Ala Asp Ser Glu Ala Pro2555 2560 2565ctc gcc gcg aac acg ccc acc cgg gcc tcg gcc gac gac cgc gag7749Leu Ala Ala Asn Thr Pro Thr Arg Ala Ser Ala Asp Asp Arg Glu2570 2575 2580ccg att gcg gtc gtt ggt atg gcc tgt cgt tat ccg ggt ggg gtg7794Pro Ile Ala Val Val Gly Met Ala Cys Arg Tyr Pro Gly Gly Val2585 2590 2595gcg gcg ccg gag gaa ctg tgg gac ctg gtg gcc gga ggc ggg cat7839Ala Ala Pro Glu Glu Leu Trp Asp Leu Val Ala Gly Gly Gly His2600 2605 2610gcg atc tcc ccg ttg cct gcc aac cga ggt tgg gac ctt gag ggg7884Ala Ile Ser Pro Leu Pro Ala Asn Arg Gly Trp Asp Leu Glu Gly2615 2620 2625ctc tac gac ccg gag ccg ggc gtg ccg ggt aag agc tat gtg cgt7929Leu Tyr Asp Pro Glu Pro Gly Val Pro Gly Lys Ser Tyr Val Arg2630 2635 2640gag ggg ggt ttt ctg cac ggg gcg gcc gag ttc gat gcg gag ttc7974Glu Gly Gly Phe Leu His Gly Ala Ala Glu Phe Asp Ala Glu Phe2645 2650 2655ttc ggt gtt tcg ccg cgt gag gcg gcg gcg atg gat ccg cag cag8019Phe Gly Val Ser Pro Arg Glu Ala Ala Ala Met Asp Pro Gln Gln2660 2665 2670cgg ttg ttg ttg gag acg tcg tgg gag gcg ttg gag cgg gcc ggg8064Arg Leu Leu Leu Glu Thr Ser Trp Glu Ala Leu Glu Arg Ala Gly2675 2680 2685atc gtg ccg gct gcg ctg cgc ggc acc cgc acc gga gtc ttc acc8109Ile Val Pro Ala Ala Leu Arg Gly Thr Arg Thr Gly Val Phe Thr2690 2695 2700ggc atc tcc cag cag gac tac gcc gcc cag ttg ggg gac gcg gcc8154Gly Ile Ser Gln Gln Asp Tyr Ala Ala Gln Leu Gly Asp Ala Ala2705 2710 2715gag acc tac ggc ggc cat gtg ctc acc gga aac ctc gga agt gtg8199Glu Thr Tyr Gly Gly His Val Leu Thr Gly Asn Leu Gly Ser Val2720 2725 2730gtc tcc ggc cgg gtt gct tac tcc ttg ggt ttg gag ggg ccg gcg8244Val Ser Gly Arg Val Ala Tyr Ser Leu Gly Leu Glu Gly Pro Ala2735 2740 2745
ctc acg gtg gat acc gcg tgt tcg tcg tcg ttg gtg gcg ttg cat8289Leu Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His2750 2755 2760ctg gcg gtg cag tcg ttg cgg cgg ggt gag tgc gat atg gcg ttg8334Leu Ala Val Gln Ser Leu Arg Arg Gly Glu Cys Asp Met Ala Leu2765 2770 2775gcc ggt ggt gtg acg gtg atg gcg acg ccg acg gtg ttt gtg gag8379Ala Gly Gly Val Thr Val Met Ala Thr Pro Thr Val Phe Val Glu2780 2785 2790ttt tcc cgg cag cgt ggg ttg gcg tcg gat ggg cgg tgc aag gcg8424Phe Ser Arg Gln Arg Gly Leu Ala Ser Asp Gly Arg Cys Lys Ala2795 2800 2805ttt gcg gag ggt gct gat ggt act gct tgg ggt gag ggt gtt ggt8469Phe Ala Glu Gly Ala Asp Gly Thr Ala Trp Gly Glu Gly Val Gly2810 2815 2820gtg ctg ttg gtg gag cgg ctg tcc gat gcc cgt cgc ctt ggt cac8514Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu Gly His2825 2830 2835tcg gtg ttg gcg gtg gtg cgg ggg agt gcg gtt aat cag gac ggt8559Ser Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly2840 2845 2850gcc agt aat ggt ttg acg gcg ccc agt ggt ccg gct cag cag agg8604Ala Ser Asn Gly Leu Thr Ala Pro Ser Gly Pro Ala Gln Gln Arg2855 2860 2865gtg atc cgt gag gcg ttg gcg gat gcc ggg ttg ggg tcg ggt gat8649Val Ile Arg Glu Ala Leu Ala Asp Ala Gly Leu Gly Ser Gly Asp2870 2875 2880gtg gat gtg gtg gag gcg cat ggt acg ggt acg gcg ttg ggt gat8694Val Asp Val Val Glu Ala His Gly Thr Gly Thr Ala Leu Gly Asp2885 2890 2895ccg atc gag gct ggt gcg ttg ctg gcc acg tat ggg cgt gag cgg8739Pro Ile Glu Ala Gly Ala Leu Leu Ala Thr Tyr Gly Arg Glu Arg2900 2905 2910gtg ggt gat ccg ttg tgg ttg ggg tcg ctg aag tcc aac atc ggg8784Val Gly Asp Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly2915 2920 2925cac act cag gcc gcc gcg ggt gtg ggt ggt gtc atc aag atg gtg8829His Thr Gln Ala Ala Ala Gly Val Gly Gly Val Ile Lys Met Val
2930 2935 2940gag gcg ctg cgt cat ggc acg ttg cct cgc act ctc cac gtc gat8874Glu Ala Leu Arg His Gly Thr Leu Pro Arg Thr Leu His Val Asp2945 2950 2955gct ccc tct tcg aag gtc gag tgg ggt tcg ggt gcg gtg gag ctg8919Ala Pro Ser Ser Lys Val Glu Trp Gly Ser Gly Ala Val Glu Leu2960 2965 2970ttg acc gag gct cga gcc tgg ccc cgg cgg gcg gat cgc aag cgc8964Leu Thr Glu Ala Arg Ala Trp Pro Arg Arg Ala Asp Arg Lys Arg2975 2980 2985cgt gcg gcc gtc tcc gcc ttc ggc gtc agc ggc acc aac gct cat9009Arg Ala Ala Val Ser Ala Phe Gly Val Ser Gly Thr Asn Ala His2990 2995 3000gtc gtc atc gag gaa ccg ccc gcc gag gtg tcg gcc gag tcg ctg9054Val Val Ile Glu Glu Pro Pro Ala Glu Val Ser Ala Glu Ser Leu3005 3010 3015gtc gag ttg cct gct ggt gct ggt gct ggt gct ggt gct ggt gct9099Val Glu Leu Pro Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Ala3020 3025 3030ggt gct ggt gct ggt gct ggg gtg tct tcg gtt gtg gcg tgg tcg9144Gly Ala Gly Ala Gly Ala Gly Val Ser Ser Val Val Ala Trp Ser3035 3040 3045ttg tcg gct cgt tcg ggt gag gcg ttg cgg gcg cag gcg gtg cgg9189Leu Ser Ala Arg Ser Gly Glu Ala Leu Arg Ala Gln Ala Val Arg3050 3055 3060ttg cgt gag cat gtg gag cgt gtt ggg gct gat ccg gtt gat gtt9234Leu Arg Glu His Val Glu Arg Val Gly Ala Asp Pro Val Asp Val3065 3070 3075gcc ttt tcg ttg gcg gtg acg cgt gcg tcg ttc ggt gag cgt gcg9279Ala Phe Ser Leu Ala Val Thr Arg Ala Ser Phe Gly Glu Arg Ala3080 3085 3090gtg gtc gtt ggt ggt gac cgt gcg gag ttg ttg gcg ggg ctg ggg9324Val Val Val Gly Gly Asp Arg Ala Glu Leu Leu Ala Gly Leu Gly3095 3100 3105gct gtt gct gct ggg gat gcg ctg tcg ggc gtg gtg cgt ggt tcg9369Ala Val Ala Ala Gly Asp Ala Leu Ser Gly Val Val Arg Gly Ser3110 3115 3120gcg gtg cgg ggg cga aag gtt gcg gct ttg ttt acg ggt cag ggt9414
Ala Val Arg Gly Arg Lys Val Ala Ala Leu Phe Thr Gly Gln Gly3125 3130 3135gcg cag tgg gtt ggt atg ggg cgt gag ttg tat ggg ttg gat ggg9459Ala Gln Trp Val Gly Met Gly Arg Glu Leu Tyr Gly Leu Asp Gly3140 3145 3150gtg ttt gct gcg gcg ttg gat gag gtt ttg ggt gtg gtg ggg gag9504Val Phe Ala Ala Ala Leu Asp Glu Val Leu Gly Val Val Gly Glu3155 3160 3165gtg ggt ggt tgg tct ttg cgt gag gtg atg ttt ggt gag ggt ggt9549Val Gly Gly Trp Ser Leu Arg Glu Val Met Phe Gly Glu Gly Gly3170 3175 3180ggt gtt ggg gtg ggg ttg ttg gat ggt acg gag ttt gcg cag cct9594Gly Val Gly Val Gly Leu Leu Asp Gly Thr Glu Phe Ala Gln Pro3185 3190 3195gct ttg ttt gcg ttg gag gtg gcg ttg ttt cgg gct gtg gag gct9639Ala Leu Phe Ala Leu Glu Val Ala Leu Phe Arg Ala Val Glu Ala3200 3205 3210cgg ggg gtg cgg gct tcg gtg gtg ttg ggg cat tcg gtg ggg gag9684Arg Gly Val Arg Ala Ser Val Val Leu Gly His Ser Val Gly Glu3215 3220 3225gtt gct gct gcg tgt gtg gcg ggg gtg ttt tcg ctt gcg gat gcg9729Val Ala Ala Ala Cys Val Ala Gly Val Phe Ser Leu Ala Asp Ala3230 3235 3240gcg cgg ttg gtg gtg gcg cgt ggt cgg ttg atg ggt ggg ttg cct9774Ala Arg Leu Val Val Ala Arg Gly Arg Leu Met Gly Gly Leu Pro3245 3250 3255gtg ggt ggg ggg atg ttg tcg gtt cgt gcg tct gag gcc gaa ctt9819Val Gly Gly Gly Met Leu Ser Val Arg Ala Ser Glu Ala Glu Leu3260 3265 3270gct gat gtt gtg gct ggg ttg ggt ggt cgg gtg tcg gtg gct gcg9864Ala Asp Val Val Ala Gly Leu Gly Gly Arg Val Ser Val Ala Ala3275 3280 3285gtc aat ggt ccg gcg tcg gtg gtg ttg tct ggt gag tgt ggt gcg9909Val Asn Gly Pro Ala Ser Val Val Leu Ser Gly Glu Cys Gly Ala3290 3295 3300ttg gat gtt gtt gcg gcg cgg ttg ggt ggg cgt ggg gtg gag tgc9954Leu Asp Val Val Ala Ala Arg Leu Gly Gly Arg Gly Val Glu Cys3305 3310 3315
aag cgg ttg gtg gtg tcg cat gcg ttt cat tcg gcg ttg atg gag9999Lys Arg Leu Val Val Ser His Ala Phe His Ser Ala Leu Met Glu3320 3325 3330ccg atg ttg gag gag ttt cgt ggg gtt gct gag agt gtg gag tat10044Pro Met Leu Glu Glu Phe Arg Gly Val Ala Glu Ser Val Glu Tyr3335 3340 3345cgg cgg ccg tgt gtg ccg gtg gtg tcg aat gtg acg ggt ggg gtg10089Arg Arg Pro Cys Val Pro Val Val Ser Asn Val Thr Gly Gly Val3350 3355 3360gtt ggg ttt gat gag ttg ggt tgt gcc gag tat tgg gtg cgg cat10134Val Gly Phe Asp Glu Leu Gly Cys Ala Glu Tyr Trp Val Arg His3365 3370 3375gcg cgg gag gcg gtg cgt ttc gct gag ggg ata cgg gct gct cgt10179Ala Arg Glu Ala Val Arg Phe Ala Glu Gly Ile Arg Ala Ala Arg3380 3385 3390gct ctt ggt gtg gat acg ttc ctg gag gtg ggt ccg cat gcg gtt10224Ala Leu Gly Val Asp Thr Phe Leu Glu Val Gly Pro His Ala Val3395 3400 3405ttg acg gcg atg gct ggt cag tgt ctt gat gga gag gag gct gac10269Leu Thr Ala Met Ala Gly Gln Cys Leu Asp Gly Glu Glu Ala Asp3410 3415 3420ttg gcg ttt gtg ccg gtc ctg cgg cgt gat cgg ccg gca ttg cag10314Leu Ala Phe Val Pro Val Leu Arg Arg Asp Arg Pro Ala Leu Gln3425 3430 3435acc ttc acc acc gca ctc gcc act ctg cac acc cgt gat gcc gaa10359Thr Phe Thr Thr Ala Leu Ala Thr Leu His Thr Arg Asp Ala Glu3440 3445 3450ctc gac gcc gtg gcg ctc cat tca ggc agc gat gcc cgg cgg atc10404Leu Asp Ala Val Ala Leu His Ser Gly Ser Asp Ala Arg Arg Ile3455 3460 3465gac ctg ccc acc tac ccc ttc caa cgc cgt agc tac tgg gcg acc10449Asp Leu Pro Thr Tyr Pro Phe Gln Arg Arg Ser Tyr Trp Ala Thr3470 3475 3480ggt tcg gtg cct ggt gcc acc ggc acc tcg gcc gcg gcc cgc ttc10494Gly Ser Val Pro Gly Ala Thr Gly Thr Ser Ala Ala Ala Arg Phe3485 3490 3495ggg ctc gta tgg aag gac cac ccg ttc ctc agc ggc gcg acg ccg10539Gly Leu Val Trp Lys Asp His Pro Phe Leu Ser Gly Ala Thr Pro3500 3505 3510
ata gcc ggc tcc gat tcg ctg ctc ctc acc ggc agg gtg gcg cct10584Ile Ala Gly Ser Asp Ser Leu Leu Leu Thr Gly Arg Val Ala Pro3515 3520 3525tcc gca tac ccg tgg ctg gcc gat cac gcc att tcc ggc acg gtg10629Ser Ala Tyr Pro Trp Leu Ala Asp His Ala Ile Ser Gly Thr Val3530 3535 3540ctg ctc cct ggg acg gcg atc gcc gac ctg ctg ctg cgg gcc gcc10674Leu Leu Pro Gly Thr Ala Ile Ala Asp Leu Leu Leu Arg Ala Ala3545 3550 3555gac gag gtg ggc gcg ggc ggt gtc gag gaa ttc atg ctc cac gcg10719Asp Glu Val Gly Ala Gly Gly Val Glu Glu Phe Met Leu His Ala3560 3565 3570ccc ctg ctc ctc ccc gaa cag ggc gga ctt cag ctc cag gtg ctg10764Pro Leu Leu Leu Pro Glu Gln Gly Gly Leu Gln Leu Gln Val Leu3575 3580 3585gtc gag gcg gcc gat gaa cga ggc tgt cgc acc gtc tcg ctc gcc10809Val Glu Ala Ala Asp Glu Arg Gly Cys Arg Thr Val Ser Leu Ala3590 3595 3600gca cgt ccc gag aat ccg ggg cgc gat ggc gag gcg ccg gag tgg10854Ala Arg Pro Glu Asn Pro Gly Arg Asp Gly Glu Ala Pro Glu Trp3605 3610 3615acc agg cac gcg gag ggt gtg ctc gcg ccc gaa ggc ccg atc gca10899Thr Arg His Ala Glu Gly Val Leu Ala Pro Glu Gly Pro Ile Ala3620 3625 3630ccg gag acc gca tgg gcc gtt ggg atc tgg ccg ccg ccc ggg gct10944Pro Glu Thr Ala Trp Ala Val Gly Ile Trp Pro Pro Pro Gly Ala3635 3640 3645gag ccg gtc gac gtc gag gag ctg tac gag ggg ttc gcc gcg gac10989Glu Pro Val Asp Val Glu Glu Leu Tyr Glu Gly Phe Ala Ala Asp3650 3655 3660ggc tac ggc tac ggc ccg gcc ttc acc gga ctg tcc ggg gtg tgg11034Gly Tyr Gly Tyr Gly Pro Ala Phe Thr Gly Leu Ser Gly Val Trp3665 3670 3675cgc cgt ggt gag gag ctc ttc gcc gag gtg cag ctg ccc gac ggg11079Arg Arg Gly Glu Glu Leu Phe Ala Glu Val Gln Leu Pro Asp Gly3680 3685 3690gtg gcg aac ggg gat aat ttc ggc att cat ccg gcc ctc ttc gac11124Val Ala Asn Gly Asp Asn Phe Gly Ile His Pro Ala Leu Phe Asp
3695 3700 3705gcg gct ctc cat cca tgg cgt gcc ggc ggg ctg gtg ccc gac acg11169Ala Ala Leu His Pro Trp Arg Ala Gly Gly Leu Val Pro Asp Thr3710 3715 3720ggc ggc acg acg ctg gtg ccg ttc tcc tgg cag ggc att ggt ctc11214Gly Gly Thr Thr Leu Val Pro Phe Ser Trp Gln Gly Ile Gly Leu3725 3730 3735cac gcc acc gga gcc gag aca ctg cgg gtc cgg ctg gcg acg gcg11259His Ala Thr Gly Ala Glu Thr Leu Arg Val Arg Leu Ala Thr Ala3740 3745 3750ggt gac ggt gcc gac gcc gcc ttc tcg gtg cag gcc gcc gac ccg11304Gly Asp Gly Ala Asp Ala Ala Phe Ser Val Gln Ala Ala Asp Pro3755 3760 3765gcc ggc cgg ccc gtc ctc acc ctg gac gcg cta ctg ctt cgc ccg11349Ala Gly Arg Pro Val Leu Thr Leu Asp Ala Leu Leu Leu Arg Pro3770 3775 3780gtg gcc ctg ggt acg gac aac gcg tcg gcg tcg ggg ctg ctg tac11394Val Ala Leu Gly Thr Asp Asn Ala Ser Ala Ser Gly Leu Leu Tyr3785 3790 3795cac gtc gac tgg cag ccg gtg ccg cgg cag gca gtt gcc ccc ggc11439His Val Asp Trp Gln Pro Val Pro Arg Gln Ala Val Ala Pro Gly3800 3805 3810tcc cgt ggc tgg acg gtt ctc ggg ccc gcc gcg agc gaa acg gcg11484Ser Arg Gly Trp Thr Val Leu Gly Pro Ala Ala Ser Glu Thr Ala3815 3820 3825acg gtg gag gtg gca cag gag gag agc gcg acc cta cga gcc ctg11529Thr Val Glu Val Ala Gln Glu Glu Ser Ala Thr Leu Arg Ala Leu3830 3835 3840ccc ggc gcg cag ccc gct gtc cac gcc gac ctc acc gct ctg cgc11574Pro Gly Ala Gln Pro Ala Val His Ala Asp Leu Thr Ala Leu Arg3845 3850 3855gcc gcc ctg gcc gcc gga acc gcc gtt ccc ggg ctg gta gtg gtg11619Ala Ala Leu Ala Ala Gly Thr Ala Val Pro Gly Leu Val Val Val3860 3865 3870ccg ccc acc ggc acc cac ctc gtc gag ccg ggc gcg ggt acg ggc11664Pro Pro Thr Gly Thr His Leu Val Glu Pro Gly Ala Gly Thr Gly3875 3880 3885ggg ggc gcg gag acg ggt gcc gca ggc tgg ggc gac gac ccc gtg11709
Gly Gly Ala Glu Thr Gly Ala Ala Gly Trp Gly Asp Asp Pro Val3890 3895 3900cgc gcc gcc ctc ggg cgc ggc ctg gcc ctg gta cgg gag tgg acc11754Arg Ala Ala Leu Gly Arg Gly Leu Ala Leu Val Arg Glu Trp Thr3905 3910 3915gag gac gaa cgc ctg gtg ggc gcc cag ctt gcc gtc ctc acc cgg11799Glu Asp Glu Arg Leu Val Gly Ala Gln Leu Ala Val Leu Thr Arg3920 3925 3930ggg gcg gtc gag gcc cgg ccc ggc gac gtg ccg gat ctg gcg ggt11844Gly Ala Val Glu Ala Arg Pro Gly Asp Val Pro Asp Leu Ala Gly3935 3940 3945gca gcc ttg tgg ggg ctg ctc cgc tcc gcg cag tcg gag tac ccc11889Ala Ala Leu Trp Gly Leu Leu Arg Ser Ala Gln Ser Glu Tyr Pro3950 3955 3960gac cgc ttc acc ctc gtc gac ctg gat gac tcc ccc gag tcc tgg11934Asp Arg Phe Thr Leu Val Asp Leu Asp Asp Ser Pro Glu Ser Trp3965 3970 3975gct gcc ctg ccc cag gct ctg gcg tcg gga gag ccg caa ctc gcc11979Ala Ala Leu Pro Gln Ala Leu Ala Ser Gly Glu Pro Gln Leu Ala3980 3985 3990ttg cgc gcc ggg acc gta ctc gct ccg gct ctc gtg ccg atc gcc12024Leu Arg Ala Gly Thr Val Leu Ala Pro Ala Leu Val Pro Ile Ala3995 4000 4005gac cct gcg acg gcc gcg acc tcg gcc gtg gcc tcg atg gcg agt12069Asp Pro Ala Thr Ala Ala Thr Ser Ala Val Ala Ser Met Ala Ser4010 4015 4020ggc gcg tcg aca gcg acc gat gtt ccc gct gcg gac gcc gca ttc12114Gly Ala Ser Thr Ala Thr Asp Val Pro Ala Ala Asp Ala Ala Phe4025 4030 4035gac ccc gac ggg acc gta ctg atc acc ggc gcc acc ggc gcc ctg12159Asp Pro Asp Gly Thr Val Leu Ile Thr Gly Ala Thr Gly Ala Leu4040 4045 4050ggg cgg cgg gtg gtc ccg cac ctg gca cgt cag cac ggc gtg cgg12204Gly Arg Arg Val Val Pro His Leu Ala Arg Gln His Gly Val Arg4055 4060 4065cat atg ctc ctg gtc agc agg cgc ggc ccg gac gcc ccc gaa gcc12249His Met Leu Leu Val Ser Arg Arg Gly Pro Asp Ala Pro Glu Ala4070 4075 4080
gcc ctc ctg gag cgg gag ctc gcc gac ctg cag gtc acc gcg acc12294Ala Leu Leu Glu Arg Glu Leu Ala Asp Leu Gln Val Thr Ala Thr4085 4090 4095ttc gcg atg tgc gac ctc gcc gac ccc gcg gac atc cgg aag gtc12339Phe Ala Met Cys Asp Leu Ala Asp Pro Ala Asp Ile Arg Lys Val4100 4105 4110atc tcc gcg gtg ccg ccg gcg cac ccg ctg acc ggt gtc gtg cac12384Ile Ser Ala Val Pro Pro Ala His Pro Leu Thr Gly Val Val His4115 4120 4125acc gcc ggc atg ctg gac gac gga gcc ctc gcc ggc ctg acg ccg12429Thr Ala Gly Met Leu Asp Asp Gly Ala Leu Ala Gly Leu Thr Pro4130 4135 4140gcg cgg ctc gat acc gtc ctc cgg ccg aaa gcc gac gcc gta cgg12474Ala Arg Leu Asp Thr Val Leu Arg Pro Lys Ala Asp Ala Val Arg4145 4150 4155aac ctg cac gag gcc act ctc gac cag ccg ttg cgc gcg ttc gtc12519Asn Leu His Glu Ala Thr Leu Asp Gln Pro Leu Arg Ala Phe Val4160 4165 4170ctg ttc tct gca gcg gcc ggg ctc ctg ggc cgc ccg ggg cag ggc12564Leu Phe Ser Ala Ala Ala Gly Leu Leu Gly Arg Pro Gly Gln Gly4175 4180 4185tcc tac gcg gcg gcc aac gcg gtc ctc gac gcg ttt gcg cgg gac12609Ser Tyr Ala Ala Ala Asn Ala Val Leu Asp Ala Phe Ala Arg Asp4190 4195 4200cgt cgt gcg gcc ggg ctg cct gct gtg tcc ctg gcc tgg gga ctg12654Arg Arg Ala Ala Gly Leu Pro Ala Val Ser Leu Ala Trp Gly Leu4205 4210 4215tgg gac gaa cgg gca ggc atg gcc ggc ggc ctg gac gac gtg gca12699Trp Asp Glu Arg Ala Gly Met Ala Gly Gly Leu Asp Asp Val Ala4220 4225 4230ctc cgt cgg ctg cgc cgc gag ggc atc gcg gcc atg ccg ccc gag12744Leu Arg Arg Leu Arg Arg Glu Gly Ile Ala Ala Met Pro Pro Glu4235 4240 4245caa gcc ctc gac ctg ctc gac ctg gcc ctg acc acg cac cgg gac12789Gln Ala Leu Asp Leu Leu Asp Leu Ala Leu Thr Thr His Arg Asp4250 4255 4260ggg ccc gcg gtc ctc gtc ccg ctc cta ctc gac ggg gcc gcc ctg12834Gly Pro Ala Val Leu Val Pro Leu Leu Leu Asp Gly Ala Ala Leu4265 4270 4275
cgc cga acg gcc aag gag cac ggc gcg acc gcg gtg cca ccg ttg12879Arg Arg Thr Ala Lys Glu His Gly Ala Thr Ala yal Pro Pro Leu4280 4285 4290ttg cgc ggc ctg ctc ccc gcg gcc ctg cgc cgc ggg agc agc ggc12924Leu Arg Gly Leu Leu Pro Ala Ala Leu Arg Arg Gly Ser Ser Gly4295 4300 4305acc ggt acc gcg gca acg gcc gcc aac cgg cgg ggc aag ggc gcg12969Thr Gly Thr Ala Ala Thr Ala Ala Asn Arg Arg Gly Lys Gly Ala4310 4315 4320gag cct gtc gcc gga cgc gtc gcg cgg atc gtg gcg ctc ctg gca13014Glu Pro Val Ala Gly Arg Val Ala Arg Ile Val Ala Leu Leu Ala4325 4330 4335gat gag agg tcc gcg gcc ctg ctg gac ctg gtc acc gag cag gtc13059Asp Glu Arg Ser Ala Ala Leu Leu Asp Leu Val Thr Glu Gln Val4340 4345 4350gcc gag gta ctc ggt cac gcg tcg gcc gcc gaa gtc gac ccc gaa13104Ala Glu Val Leu Gly His Ala Ser Ala Ala Glu Val Asp Pro Glu4355 4360 4365cgt ccc ttc cgg gac atc ggc ttc gac tcc ctg gcg gcg gtg gag13149Arg Pro Phe Arg Asp Ile Gly Phe Asp Ser Leu Ala Ala Val Glu4370 4375 4380ctg cgc aac cgc ctc ggc cgc ctg gtc gac ctg cgg ctg ccg acc13194Leu Arg Asn Arg Leu Gly Arg Leu Val Asp Leu Arg Leu Pro Thr4385 4390 4395aca ctc gcc ttc gac cgc ccc acg ccg aag gac gtg gcc gag tgg13239Thr Leu Ala Phe Asp Arg Pro Thr Pro Lys Asp Val Ala Glu Trp4400 4405 4410ctc gac ggg gag ttg ccc cgc ccc gcc ggt tcg tca gcc gat tcc13284Leu Asp Gly Glu Leu Pro Arg Pro Ala Gly Ser Ser Ala Asp Ser4415 4420 4425tcc gcg ctg gag ggg atc gac gaa ctc gcc cgg gcc gtc gcc ctg13329Ser Ala Leu Glu Gly Ile Asp Glu Leu Ala Arg Ala Val Ala Leu4430 4435 4440ctg ggc ccg gac gac gcc cgg cga gcc gag gta cgg cag cgg ctc13374Leu Gly Pro Asp Asp Ala Arg Arg Ala Glu Val Arg Gln Arg Leu4445 4450 4455act ggg ctg ctg gcc gag ctc gac acc ccc ggg cac ggc act gcc13419Thr Gly Leu Leu Ala Glu Leu Asp Thr Pro Gly His Gly Thr Ala
4460 4465 4470ggc ccc cga gac cgc acc gcc ccc gcc gat gcc gag agc acc ccg13464Gly Pro Arg Asp Arg Thr Ala Pro Ala Asp Ala Glu Ser Thr Pro4475 4480 4485gcg act gtg gcg ggc cgg ctt gac gag gcg act gac gac gag atc13509Ala Thr Val Ala Gly Arg Leu Asp Glu Ala Thr Asp Asp Glu Ile4490 4495 4500ttc gcc ttc ctg gac gag cag ctg tga 13536Phe Ala Phe Leu Asp Glu Gln Leu4505 4510<210>4<211>4511<212>PRT<213>Streptomyces mycarofaciens<400>4Met Leu Val Ser Gly Asp Leu Val Thr Ser Arg Ile Asp Asp Arg Ser1 5 10 15Asp Ala Ile Ala Val Val Gly Met Ser Cys Arg Phe Pro Gly Ala Pro20 25 30Gly Val Glu Glu Phe Trp Lys Leu Leu Thr Asp Gly Thr Glu Ala Val35 40 45Ser Arg Ala Ala Asp Gly Arg Arg Arg Gly Met Ile Glu Ala Val Gly50 55 60Asp Phe Asp Ala Thr Phe Phe Gly Met Ser Pro Arg Glu Ala Ala Glu65 70 75 80Thr Asp Pro Gln Gln Arg Leu Leu Leu Glu Leu Gly Trp Glu Ala Leu85 90 95Glu Asp Ala Gly Ile Val Pro Gly Ser Leu Arg Gly Glu Ala Val Gly100 105 110Ile Phe Val Gly Ala Met His Asn Asp Tyr Ala Thr Leu Leu His Arg
115 120 125Ala Gly Ala Pro Ala Gly Ala His Thr Ala Thr Gly Leu Gln Pro Ala130 135 140Met Leu Ala Asn Arg Leu Ser Tyr Val Leu Gly Thr Arg Gly Pro Ser145 150 155 160Leu Ala Val Asp Thr Ala Gln Ser Ser Ser Leu Val Ala Val Ala Leu165 170 175Ala Val Glu Ser Leu Arg Ala Gly Thr Ser Arg Ile Ala Ile Ala Gly180 185 190Gly Val Asn Leu Ile Leu Ala Asp Glu Gly Ser Ala Thr Met Glu Arg195 200 205Leu Gly Ala Leu Ser Pro Asp Gly Arg Cys Tyr Thr Phe Asp Ala Arg210 215 220Ala Asn Gly Tyr Val Arg Gly Glu Gly Gly Ala Ala Val Val Leu Lys225 230 235 240Pro Leu Ala Asp Ala Leu Ala Asp Gly Asp Pro Val Tyr Cys Val Val245 250 255Arg Ser Ala Ala Thr Gly Asn Asp Gly Gly Gly Pro Gly Leu Thr Ser260 265 270Pro Asp His Glu Gly Gln Glu Ala Val Leu Arg Ala Ala Cys Ala Gln275 280 285Ala Gly Val Asp Pro Ala Lys Val Arg Phe Val Glu Leu His Gly Thr290 295 300Gly Thr Pro Val Gly Asp Pro Val Glu Ala Arg Ala Leu Gly Ala Val305 310 315 320
His Gly Ser Gly Arg Pro Ala Asp Ala Pro Leu Leu Val Gly Ser Val325 330 335Lys Thr Asn Ile Gly His Leu Glu Gly Ala Ala Gly Ile Ala Gly Leu340 345 350Val Lys Ala Ala Leu Cys Leu Arg Asn Arg Thr Leu Pro Gly Ser Leu355 360 365Asn Phe Val Thr Pro His Pro Ala Ile Pro Leu Asp Arg Leu Arg Leu370 375 380Lys Val Gln Thr Thr Pro Thr Thr Leu His Pro Asp Pro Asp Gly Ser385 390 395 400Pro Leu Leu Ala Gly Val Ser Ser Phe Gly Ile Gly Gly Thr Asn Cys405 410 415His Val Val Leu Glu His Leu Pro Glu Pro Ala Pro Thr Thr Arg Glu420 425 430Ala Leu Pro Ala Pro His Leu Val Pro Pro Leu Leu Leu Ser Ala Arg435 440 445Ser His Pro Ala Leu Leu Ala Gln Ala Ala Arg Leu Arg Asp His Leu450 455 460Ser Arg Thr Ala Ala Asp Pro Gln Asp Val Ala Tyr Ser Leu Ala Thr465 470 475 480Thr Arg Ser Leu Phe Glu His Arg Ala Ala Leu Pro Cys Gly Asn Arg485 490 495Glu Glu Leu Val Ala Ala Leu Asp Ala Leu Ala His Gly Arg Ile Thr500 505 510Ala Gly Val Arg Val Asp Ser Ala Val Ser Gly Gly Thr Ala Val Leu515 520 525
Phe Thr Gly Gln Gly Ala Gln Trp Val Gly Met Gly Arg Glu Leu Tyr530 535 540Gly Leu Asp Gly Val Phe Ala Ala Ala Leu Asp Glu Val Leu Gly Val545 550 555 560Val Gly Glu Val Gly Gly Trp Ser Leu Arg Glu Val Met Phe Gly Glu565 570 575Gly Gly Gly Val Gly Val Gly Leu Leu Asp Gly Thr Glu Phe Ala Gln580 585 590Pro Ala Leu Phe Ala Leu Glu Val Ala Leu Phe Arg Ala Val Glu Ala595 600 605Arg Gly Val Arg Ala Ser Val Val Leu Gly His Ser Val Gly Glu Val610 615 620Ala Ala Ala Cys Val Ala Gly Val Phe Ser Leu Ala Asp Ala Ala Arg625 630 635 640Leu Val Val Ala Arg Gly Arg Leu Met Gly Ala Leu Pro Val Gly Gly645 650 655Gly Met Leu Ser Val Arg Ala Ser Glu Ala Glu Leu Val Asp Val Val660 665 670Ala Gly Leu Gly Gly Arg Val Ser Val Ala Ala Val Asn Gly Pro Ala675 680 685Ser Val Val Leu Ser Gly Glu Cys Gly Ala Leu Asp Val Val Ala Ala690 695 700Arg Leu Gly Gly Arg Gly Val Glu Cys Lys Arg Leu Val Val Ser His705 710 715 720Ala Phe His Ser Ala Leu Met Asp Pro Met Leu Glu Glu Phe Arg Gly725 730 735
Val Ala Glu Ser Val Glu Tyr Arg Arg Pro Cys Val Pro Val Val Ser740 745 750Asn Val Thr Gly Gly Val Val Gly Phe Asp Glu Leu Gly Cys Ala Glu755 760 765Tyr Trp Val Arg His Ala Arg Glu Ala Val Arg Phe Ala Glu Gly Ile770 775 780Arg Ala Ala Arg Ala Leu Gly Val Asp Thr Phe Leu Glu Val Gly Pro785 790 795 800His Ala Val Leu Thr Ala Met Ala Gly Gln Cys Leu Asp Ala Glu Glu805 810 815Ala Asp Leu Ala Phe Val Pro Val Leu Arg Arg Asp Arg Pro Ala Leu820 825 830Gln Thr Phe Thr Thr Ala Leu Ala Thr Leu His Thr Arg Asp Ala Glu835 840 845Leu Asp Ala Val Ala Leu His Ser Gly Ser Asp Ala Arg Arg Ile Asp850 855 860Leu Pro Thr Tyr Pro Phe Gln Arg Arg Thr His Trp Ser Pro Ala Leu865 870 875 880Ser His Gly His Ala Ala Gly Val Val Arg Ala Ser Thr Ala Thr Glu885 890 895Ile Arg Gly Asn Asp Glu Ile Pro Glu Ser Ala Glu Ala Leu Leu Arg900 905 910Asp Pro Ala Asp Gly Ser Leu Ala Ala Ser Pro Glu Pro Ala Thr Pro915 920 925Asp Gln Leu Val Arg Leu Val Arg Glu Thr Thr Ala Ala Val Leu Gly
930 935 940His Asp Asp Pro Asp Glu Ile Val Leu Asp Arg Thr Phe Thr Ser Gln945 950 955 960Gly Leu Glu Ser Val Thr Ala Val Glu Leu Arg Asp Leu Leu Asn Arg965 970 975Ala Thr Gly Leu Thr Leu Ala Ala Thr Leu Val Tyr Asp Leu Pro Thr980 985 990Pro Arg Ala Val Ala Asp Tyr Leu Ser Ala Ala Met Leu Ala Thr Asp995 1000 1005Asp Ala Asn Ser Ser Ala His Gln Thr Thr Ala Ala Ala Thr Thr1010 1015 1020Arg Ser Gly Ala Arg Asn Asp Asp Pro Ile Ala Ile Val Gly Val1025 1030 1035Gly Ser His Phe Pro Gly Gly Val Asp Ser Arg Ala Gly Leu Trp1040 1045 1050Asp Leu Leu Ala Ser Gly Thr Asp Ala Ile Ser Ser Phe Pro Thr1055 1060 1065Asp Arg Gly Trp Asp Leu Asn Glu Leu Tyr Asp Pro Glu Pro Gly1070 1075 1080Ile Pro Gly Lys Thr Tyr Val Arg Gln Gly Gly Phe Leu His Gln1085 1090 1095Ala Ala Glu Phe Asp Ala Glu Phe Phe Gly Ile Ser Pro Arg Glu1100 1105 1110Ala Thr Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Thr Ser1115 1120 1125
Trp Glu Ala Leu Glu Asp Ala Gly Val Cys Pro Glu Ser Leu Arg1130 1135 1140Gly Thr Asn Thr Gly Val Phe Ile Gly Ala Val Ala Pro Glu Tyr1145 1150 1155Gly Pro Arg Leu His Glu Gly Ala Asp Gly Tyr Glu Gly Tyr Leu1160 1165 1170Leu Thr Gly Thr Thr Ala Ser Val Ala Ser Gly Arg Ile Ala Tyr1175 1180 1185Thr Phe Gly Thr Arg Gly Pro Ala Leu Thr Val Asp Thr Ala Cys1190 1195 1200Ser Ser Ser Leu Val Ala Leu His Leu Ala Val Gln Ser Leu Arg1205 1210 1215Arg Gly Glu Cys Asp Met Ala Leu Ala Gly Gly Ala Thr Val Met1220 1225 1230Ser Gly Pro Gly Met Phe Val Glu Phe Ser Arg Gln Arg Gly Leu1235 1240 1245Ala Ser Asp Gly Arg Cys Lys Ala Phe Ser Ala Asp Ala Asp Gly1250 1255 1260Thr Ala Trp Ser Glu Gly Val Ala Val Leu Ala Leu Glu Arg Leu1265 1270 1275Ser Asp Ala Arg Arg Ala Gly His Arg Val Leu Ala Leu Val Arg1280 1285 1290Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala1295 1300 1305Pro Ser Gly Pro Ala Gln Glu Ser Val Ile Arg Glu Ala Leu Ala1310 1315 1320
Asp Ala Gly Leu Gly Pro Gly Asp Val Asp Val Val Glu Ala His1325 1330 1335Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile Glu Ala Gly Ala Leu1340 1345 1350Leu Ala Thr Tyr Gly Cys Glu Arg Val Gly Asp Pro Leu Trp Leu1355 1360 1365Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gln Ala Ala Ala Gly1370 1375 1380Val Ala Gly Val Ile Lys Met Val Glu Ala Leu Arg His Gly Thr1385 1390 1395Leu Pro Arg Thr Leu His Ala Asp Arg Pro Ser Thr His Val Asp1400 1405 1410Trp Ser Ser Gly Gly Val Glu Leu Leu Thr Glu Ala Arg Pro Trp1415 1420 1425Pro Glu Arg Glu Gly Arg Pro Arg Arg Ala Ala Val Ser Ala Phe1430 1435 1440Gly Val Ser Gly Thr Asn Ala His Leu Val Ile Glu Glu Pro Pro1445 1450 1455Val Glu Leu Pro Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Ala1460 1465 1470Gly Val Ser Ser Val Val Ala Trp Pro Leu Ser Ala Arg Ser Gly1475 1480 1485Glu Ala Leu Arg Ala Gln Ala Val Arg Leu Arg Glu His Val Glu1490 1495 1500Arg Val Gly Ala Asp Pro Val Asp Val Ala Phe Ser Leu Ala Val1505 1510 1515
Thr Arg Ala Ser Phe Gly Glu Arg Ala Val Val Val Gly Gly Asp1520 1525 1530Arg Ala Glu Leu Leu Ala Gly Leu Asp Ala Leu Ala Gly Gly Arg1535 1540 1545Arg Gly Pro Gly Val Val Arg Gly Ser Ala Val Ser Gly Gly Thr1550 1555 1560Ala Val Leu Phe Thr Gly Gln Gly Ala Gln Trp Val Gly Met Gly1565 1570 1575Arg Glu Leu Tyr Gly Leu Asp Gly Val Phe Ala Ala Ala Leu Asp1580 1585 1590Glu Val Leu Gly Val Val Gly Glu Val Gly Gly Trp Ser Leu Arg1595 1600 1605Glu Val Met Phe Gly Glu Gly Gly Gly Val Gly Val Gly Leu Leu1610 1615 1620Asp Gly Thr Glu Phe Ala Gln Pro Ala Leu Phe Ala Leu Glu Val1625 1630 1635Ala Leu Phe Arg Ala Val Glu Ala Arg Gly Val Arg Ala Ser Val1640 1645 1650Val Leu Gly His Ser Val Gly Glu Val Ala Ala Ala Cys Val Ala1655 1660 1665Gly Val Phe Ser Leu Ala Asp Ala Ala Arg Leu Val Val Ala Arg1670 1675 1680Gly Arg Leu Met Gly Gly Leu Pro Val Gly Gly Gly Met Leu Ser1685 1690 1695Val Arg Ala Ser Glu Ala Glu Leu Ala Asp Val Val Ala Gly Leu
1700 1705 1710Gly Gly Arg Val Ser Val Ala Ala Val Asn Gly Pro Ala Ser Val1715 1720 1725Val Leu Ser Gly Glu Cys Gly Ala Leu Asp Val Val Ala Ala Arg1730 1735 1740Leu Gly Gly Arg Gly Val Glu Cys Lys Arg Leu Val Val Ser His1745 1750 1755Ala Phe His Ser Ala Leu Met Glu Pro Met Leu Glu Glu Phe Arg1760 1765 1770Gly Val Ala Glu Ser Val Glu Tyr Arg Arg Pro Cys Val Pro Val1775 1780 1785Val Ser Asn Val Thr Gly Gly Val Val Gly Phe Asp Glu Leu Gly1790 1795 1800Cys Ala Glu Tyr Trp Val Arg His Ala Arg Glu Ala Val Arg Phe1805 1810 1815Ala Glu Gly Ile Arg Ala Ala Arg Ala Leu Gly Val Asp Thr Phe1820 1825 1830Leu Glu Val Gly Pro His Ala Val Leu Thr Ala Met Ala Gly Gln1835 1840 1845Cys Leu Asp Gly Glu Glu Ala Asp Leu Ala Phe Val Pro Val Leu1850 1855 1860Arg Arg Asp Arg Pro Ala Ser Gln Thr Phe Thr Thr Ala Leu Ala1865 1870 1875Thr Leu Cys Val Arg Gly Thr Glu Val Asp Trp Ala Thr Pro His1880 1885 1890
Arg Lys Ser Gly Ala Gln Arg Ile Asp Leu Pro Thr Tyr Pro Phe1895 1900 1905Gln Arg Ala Arg Tyr Trp Leu Asp Pro Ala Pro Ala Met Ala Leu1910 1915 1920Thr Thr Val Ala Ala Ser Ser Ala Glu Ala Ala Ala Thr Ala Thr1925 1930 1935Glu Gly Thr Ala Leu Glu Thr Ala Gly Leu Arg Tyr Arg Ile Ala1940 1945 1950Trp Gln Ala Ala Ala Thr Asp Arg Gly Thr Ser Arg Ser Ala Gly1955 1960 1965His Val Val Leu Leu Thr Ser Asp Asp Asp Ala Thr Glu Ser Gly1970 1975 1980Leu Ala Ala Ala Ile Thr Arg Glu Leu Ala Val Arg Gly Ala Glu1985 1990 1995Val Arg Thr Ala Ile Leu Pro Val Gly Thr Asp Arg Glu Thr Ala2000 2005 2010Ala Asp Leu Leu Arg Thr Ser Gly Asp Gly Ala Ala Arg Ser Thr2015 2020 2025His Val Leu Trp Leu Ala Pro Ala Glu Pro Asp Thr Ala Asp Ala2030 2035 2040Val Ala Leu Ile Gln Ala Leu Gly Glu Ala Gly His Asp Ala Pro2045 2050 2055Leu Trp Ile Ala Thr Arg Asp Ala Val Ala Val Gln Pro Gly Glu2060 2065 2070Lys Leu Ser Val Ala Gly Ala Gln Leu Trp Gly Leu Gly Gln Val2075 2080 2085
Ala Ala Leu Glu Leu Phe Gln Arg Trp Gly Gly Leu Val Asp Leu2090 2095 2100Pro Glu Asn Pro Ser Pro Ala Ala Val Arg Ala Phe Val Gly Ala2105 2110 2115Leu Phe Ala Glu Gly Asp Asp Asn Gln Ile Ala Val Arg Pro Ser2120 2125 2130Gly Val Tyr Val Arg Arg Val Ala Pro Ala Pro Ala Pro Ala Pro2135 2140 2145Ala Leu Ile Gly Gln Ala Ala Gln Asp Asp Arg Ser Gly Pro Ser2150 2155 2160Asp Gly Leu Asp Gly Asn Asn Gly Thr Ala Pro Val Asn Trp His2165 2170 2175Pro Ser Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ala Leu Gly2180 2185 2190Ala Gln Val Ala Arg Arg Leu Ala Arg Ala Gly Ala Pro His Leu2195 2200 2205Leu Leu Val Ser Arg Arg Gly Pro Asp Gly Pro Gly Thr Gly Glu2210 2215 2220Leu Val Gly Glu Leu Thr Ala His Gly Thr Glu Val Thr Val Thr2225 2230 2235Ala Cys Asp Ala Ala Asp Arg Asp Ala Leu Ala Glu Leu Leu Ala2240 2245 2250Ser Ile Pro Glu Asp Arg Pro Leu Thr Ala Val Leu His Ala Ala2255 2260 2265Gly Val Leu Asp Asp Gly Val Leu Asp Ala Leu Thr Pro Asp Arg2270 2275 2280
Leu Asp Ala Val Leu Arg Ala Lys Val Thr Val Ala Arg His Leu2285 2290 2295Asp Glu Leu Thr Ala Gly Ile Pro Leu Asp Ala Phe Val Leu Phe2300 2305 2310Ser Ser Ile Val Gly Val Trp Gly Asn Gly Gly Gln Gly Gly Tyr2315 2320 2325Ala Ala Ala Asn Ala Ala Leu Asp Ala Leu Ala His Arg Arg Arg2330 2335 2340Ala Arg Gly Gln Arg Ala Thr Ser Ile Ala Trp Gly Pro Trp Ala2345 2350 2355Gly Ala Gly Met Ala Ala Gly Ala Gly Ser Lys Ala Phe Gln Arg2360 2365 2370Asp Gly Ile Gln Ala Leu Asp Pro Glu Arg Ala Leu Asn Val Leu2375 2380 2385Asp Asp Val Val Arg Ala Asp Glu Thr Ser Val Ala Ala Glu Pro2390 2395 2400Ser Leu Ile Val Ala Asp Val Asp Trp Ser Thr Phe Val Gly Arg2405 2410 2415Ser Val Ala Arg Arg Thr Trp Ala Leu Phe Asp Gly Val Pro Ala2420 2425 2430Ala Cys Ser Ala Arg Ser Ala Gln Ala Ala Gln Gly Arg Ser Ala2435 2440 2445His Ala Pro Gly Glu Arg Pro His His Gly Gly Ile Gly Gly Ser2450 2455 2460Gly Asp Gly Ala Asp Glu Asp Arg Pro Trp Leu Ser Ala Gly Pro
2465 2470 2475Ser Ser Pro Glu Arg Arg Arg Ala Leu Leu Asp Leu Val Arg Ser2480 2485 2490Glu Ala Ala Glu Ile Leu Arg His Gly Ser Ala Ala Ala Val Asp2495 2500 2505Pro Glu Thr Ala Phe Arg Ala Ala Gly Phe Asp Ser Leu Thr Val2510 2515 2520Leu Glu Leu Arg Asn Arg Leu Thr Ala Ala Ile Gly Leu Asn Leu2525 2530 2535Pro Ser Thr Leu Leu Phe Asp Tyr Pro Asn Pro Asn Ala Leu Ala2540 2545 2550Asp His Leu His Asp Glu Leu Phe Gly Ala Asp Ser Glu Ala Pro2555 2560 2565Leu Ala Ala Asn Thr Pro Thr Arg Ala Ser Ala Asp Asp Arg Glu2570 2575 2580Pro Ile Ala Val Val Gly Met Ala Cys Arg Tyr Pro Gly Gly Val2585 2590 2595Ala Ala Pro Glu Glu Leu Trp Asp Leu Val Ala Gly Gly Gly His2600 2605 2610Ala Ile Ser Pro Leu Pro Ala Asn Arg Gly Trp Asp Leu Glu Gly2615 2620 2625Leu Tyr Asp Pro Glu Pro Gly Val Pro Gly Lys Ser Tyr Val Arg2630 2635 2640Glu Gly Gly Phe Leu His Gly Ala Ala Glu Phe Asp Ala Glu Phe2645 2650 2655
Phe Gly Val Ser Pro Arg Glu Ala Ala Ala Met Asp Pro Gln Gln2660 2665 2670Arg Leu Leu Leu Glu Thr Ser Trp Glu Ala Leu Glu Arg Ala Gly2675 2680 2685Ile Val Pro Ala Ala Leu Arg Gly Thr Arg Thr Gly Val Phe Thr2690 2695 2700Gly Ile Ser Gln Gln Asp Tyr Ala Ala Gln Leu Gly Asp Ala Ala2705 2710 2715Glu Thr Tyr Gly Gly His Val Leu Thr Gly Asn Leu Gly Ser Val2720 2725 2730Val Ser Gly Arg Val Ala Tyr Ser Leu Gly Leu Glu Gly Pro Ala2735 2740 2745Leu Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His2750 2755 2760Leu Ala Val Gln Ser Leu Arg Arg Gly Glu Cys Asp Met Ala Leu2765 2770 2775Ala Gly Gly Val Thr Val Met Ala Thr Pro Thr Val Phe Val Glu2780 2785 2790Phe Ser Arg Gln Arg Gly Leu Ala Ser Asp Gly Arg Cys Lys Ala2795 2800 2805Phe Ala Glu Gly Ala Asp Gly Thr Ala Trp Gly Glu Gly Val Gly2810 2815 2820Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu Gly His2825 2830 2835Ser Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly2840 2845 2850
Ala Ser Asn Gly Leu Thr Ala Pro Ser Gly Pro Ala Gln Gln Arg2855 2860 2865Val Ile Arg Glu Ala Leu Ala Asp Ala Gly Leu Gly Ser Gly Asp2870 2875 2880Val Asp Val Val Glu Ala His Gly Thr Gly Thr Ala Leu Gly Asp2885 2890 2895Pro Ile Glu Ala Gly Ala Leu Leu Ala Thr Tyr Gly Arg Glu Arg2900 2905 2910Val Gly Asp Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly2915 2920 2925His Thr Gln Ala Ala Ala Gly Val Gly Gly Val Ile Lys Met Val2930 2935 2940Glu Ala Leu Arg His Gly Thr Leu Pro Arg Thr Leu His Val Asp2945 2950 2955Ala Pro Ser Ser Lys Val Glu Trp Gly Ser Gly Ala Val Glu Leu2960 2965 2970Leu Thr Glu Ala Arg Ala Trp Pro Arg Arg Ala Asp Arg Lys Arg2975 2980 2985Arg Ala Ala Val Ser Ala Phe Gly Val Ser Gly Thr Asn Ala His2990 2995 3000Val Val Ile Glu Glu Pro Pro Ala Glu Val Ser Ala Glu Ser Leu3005 3010 3015Val Glu Leu Pro Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Ala3020 3025 3030Gly Ala Gly Ala Gly Ala Gly Val Ser Ser Val Val Ala Trp Ser3035 3040 3045
Leu Ser Ala Arg Ser Gly Glu Ala Leu Arg Ala Gln Ala Val Arg3050 3055 3060Leu Arg Glu His Val Glu Arg Val Gly Ala Asp Pro Val Asp Val3065 3070 3075Ala Phe Ser Leu Ala Val Thr Arg Ala Ser Phe Gly Glu Arg Ala3080 3085 3090Val Val Val Gly Gly Asp Arg Ala Glu Leu Leu Ala Gly Leu Gly3095 3100 3105Ala Val Ala Ala Gly Asp Ala Leu Ser Gly Val Val Arg Gly Ser3110 3115 3120Ala Val Arg Gly Arg Lys Val Ala Ala Leu Phe Thr Gly Gln Gly3125 3130 3135Ala Gln Trp Val Gly Met Gly Arg Glu Leu Tyr Gly Leu Asp Gly3140 3145 3150Val Phe Ala Ala Ala Leu Asp Glu Val Leu Gly Val Val Gly Glu3155 3160 3165Val Gly Gly Trp Ser Leu Arg Glu Val Met Phe Gly Glu Gly Gly3170 3175 3180Gly Val Gly Val Gly Leu Leu Asp Gly Thr Glu Phe Ala Gln Pro3185 3190 3195Ala Leu Phe Ala Leu Glu Val Ala Leu Phe Arg Ala Val Glu Ala3200 3205 3210Arg Gly Val Arg Ala Ser Val Val Leu Gly His Ser Val Gly Glu3215 3220 3225Val Ala Ala Ala Cys Val Ala Gly Val Phe Ser Leu Ala Asp Ala
3230 3235 3240Ala Arg Leu Val Val Ala Arg Gly Arg Leu Met Gly Gly Leu Pro3245 3250 3255Val Gly Gly Gly Met Leu Ser Val Arg Ala Ser Glu Ala Glu Leu3260 3265 3270Ala Asp Val Val Ala Gly Leu Gly Gly Arg Val Ser Val Ala Ala3275 3280 3285Val Asn Gly Pro Ala Ser Val Val Leu Ser Gly Glu Cys Gly Ala3290 3295 3300Leu Asp Val Val Ala Ala Arg Leu Gly Gly Arg Gly Val Glu Cys3305 3310 3315Lys Arg Leu Val Val Ser His Ala Phe His Ser Ala Leu Met Glu3320 3325 3330Pro Met Leu Glu Glu Phe Arg Gly Val Ala Glu Ser Val Glu Tyr3335 3340 3345Arg Arg Pro Cys Val Pro Val Val Ser Asn Val Thr Gly Gly Val3350 3355 3360Val Gly Phe Asp Glu Leu Gly Cys Ala Glu Tyr Trp Val Arg His3365 3370 3375Ala Arg Glu Ala Val Arg Phe Ala Glu Gly Ile Arg Ala Ala Arg3380 3385 3390Ala Leu Gly Val Asp Thr Phe Leu Glu Val Gly Pro His Ala Val3395 3400 3405Leu Thr Ala Met Ala Gly Gln Cys Leu Asp Gly Glu Glu Ala Asp3410 3415 3420
Leu Ala Phe Val Pro Val Leu Arg Arg Asp Arg Pro Ala Leu Gln3425 3430 3435Thr Phe Thr Thr Ala Leu Ala Thr Leu His Thr Arg Asp Ala Glu3440 3445 3450Leu Asp Ala Val Ala Leu His Ser Gly Ser Asp Ala Arg Arg Ile3455 3460 3465Asp Leu Pro Thr Tyr Pro Phe Gln Arg Arg Ser Tyr Trp Ala Thr3470 3475 3480Gly Ser Val Pro Gly Ala Thr Gly Thr Ser Ala Ala Ala Arg Phe3485 3490 3495Gly Leu Val Trp Lys Asp His Pro Phe Leu Ser Gly Ala Thr Pro3500 3505 3510Ile Ala Gly Ser Asp Ser Leu Leu Leu Thr Gly Arg Val Ala Pro3515 3520 3525Ser Ala Tyr Pro Trp Leu Ala Asp His Ala Ile Ser Gly Thr Val3530 3535 3540Leu Leu Pro Gly Thr Ala Ile Ala Asp Leu Leu Leu Arg Ala Ala3545 3550 3555Asp Glu Val Gly Ala Gly Gly Val Glu Glu Phe Met Leu His Ala3560 3565 3570Pro Leu Leu Leu Pro Glu Gln Gly Gly Leu Gln Leu Gln Val Leu3575 3580 3585Val Glu Ala Ala Asp Glu Arg Gly Cys Arg Thr Val Ser Leu Ala3590 3595 3600Ala Arg Pro Glu Asn Pro Gly Arg Asp Gly Glu Ala Pro Glu Trp3605 3610 3615
Thr Arg His Ala Glu Gly Val Leu Ala Pro Glu Gly Pro Ile Ala3620 3625 3630Pro Glu Thr Ala Trp Ala Val Gly Ile Trp Pro Pro Pro Gly Ala3635 3640 3645Glu Pro Val Asp Val Glu Glu Leu Tyr Glu Gly Phe Ala Ala Asp3650 3655 3660Gly Tyr Gly Tyr Gly Pro Ala Phe Thr Gly Leu Ser Gly Val Trp3665 3670 3675Arg Arg Gly Glu Glu Leu Phe Ala Glu Val Gln Leu Pro Asp Gly3680 3685 3690Val Ala Asn Gly Asp Asn Phe Gly Ile His Pro Ala Leu Phe Asp3695 3700 3705Ala Ala Leu His Pro Trp Arg Ala Gly Gly Leu Val Pro Asp Thr3710 3715 3720Gly Gly Thr Thr Leu Val Pro Phe Ser Trp Gln Gly Ile Gly Leu3725 3730 3735His Ala Thr Gly Ala Glu Thr Leu Arg Val Arg Leu Ala Thr Ala3740 3745 3750Gly Asp Gly Ala Asp Ala Ala Phe Ser Val Gln Ala Ala Asp Pro3755 3760 3765Ala Gly Arg Pro Val Leu Thr Leu Asp Ala Leu Leu Leu Arg Pro3770 3775 3780Val Ala Leu Gly Thr Asp Asn Ala Ser Ala Ser Gly Leu Leu Tyr3785 3790 3795His Val Asp Trp Gln Pro Val Pro Arg Gln Ala Val Ala Pro Gly3800 3805 3810
Ser Arg Gly Trp Thr Val Leu Gly Pro Ala Ala Ser Glu Thr Ala3815 3820 3825Thr Val Glu Val Ala Gln Glu Glu Ser Ala Thr Leu Arg Ala Leu3830 3835 3840Pro Gly Ala Gln Pro Ala Val His Ala Asp Leu Thr Ala Leu Arg3845 3850 3855Ala Ala Leu Ala Ala Gly Thr Ala Val Pro Gly Leu Val Val Val3860 3865 3870Pro Pro Thr Gly Thr His Leu Val Glu Pro Gly Ala Gly Thr Gly3875 3880 3885Gly Gly Ala Glu Thr Gly Ala Ala Gly Trp Gly Asp Asp Pro Val3890 3895 3900Arg Ala Ala Leu Gly Arg Gly Leu Ala Leu Val Arg Glu Trp Thr3905 3910 3915Glu Asp Glu Arg Leu Val Gly Ala Gln Leu Ala Val Leu Thr Arg3920 3925 3930Gly Ala Val Glu Ala Arg Pro Gly Asp Val Pro Asp Leu Ala Gly3935 3940 3945Ala Ala Leu Trp Gly Leu Leu Arg Ser Ala Gln Ser Glu Tyr Pro3950 3955 3960Asp Arg Phe Thr Leu Val Asp Leu Asp Asp Ser Pro Glu Ser Trp3965 3970 3975Ala Ala Leu Pro Gln Ala Leu Ala Ser Gly Glu Pro Gln Leu Ala3980 3985 3990Leu Arg Ala Gly Thr Val Leu Ala Pro Ala Leu Val Pro Ile Ala
3995 4000 4005Asp Pro Ala Thr Ala Ala Thr Ser Ala Val Ala Ser Met Ala Ser4010 4015 4020Gly Ala Ser Thr Ala Thr Asp Val Pro Ala Ala Asp Ala Ala Phe4025 4030 4035Asp Pro Asp Gly Thr Val Leu Ile Thr Gly Ala Thr Gly Ala Leu4040 4045 4050Gly Arg Arg Val Val Pro His Leu Ala Arg Gln His Gly Val Arg4055 4060 4065His Met Leu Leu Val Ser Arg Arg Gly Pro Asp Ala Pro Glu Ala4070 4075 4080Ala Leu Leu Glu Arg Glu Leu Ala Asp Leu Gln Val Thr Ala Thr4085 4090 4095Phe Ala Met Cys Asp Leu Ala Asp Pro Ala Asp Ile Arg Lys Val4100 4105 4110Ile Ser Ala Val Pro Pro Ala His Pro Leu Thr Gly Val Val His4115 4120 4125Thr Ala Gly Met Leu Asp Asp Gly Ala Leu Ala Gly Leu Thr Pro4130 4135 4140Ala Arg Leu Asp Thr Val Leu Arg Pro Lys Ala Asp Ala Val Arg4145 4150 4155Asn Leu His Glu Ala Thr Leu Asp Gln Pro Leu Arg Ala Phe Val4160 4165 4170Leu Phe Ser Ala Ala Ala Gly Leu Leu Gly Arg Pro Gly Gln Gly4175 4180 4185
Ser Tyr Ala Ala Ala Asn Ala Val Leu Asp Ala Phe Ala Arg Asp4190 4195 4200Arg Arg Ala Ala Gly Leu Pro Ala Val Ser Leu Ala Trp Gly Leu4205 4210 4215Trp Asp Glu Arg Ala Gly Met Ala Gly Gly Leu Asp Asp Val Ala4220 4225 4230Leu Arg Arg Leu Arg Arg Glu Gly Ile Ala Ala Met Pro Pro Glu4235 4240 4245Gln Ala Leu Asp Leu Leu Asp Leu Ala Leu Thr Thr His Arg Asp4250 4255 4260Gly Pro Ala Val Leu Val Pro Leu Leu Leu Asp Gly Ala Ala Leu4265 4270 4275Arg Arg Thr Ala Lys Glu His Gly Ala Thr Ala Val Pro Pro Leu4280 4285 4290Leu Arg Gly Leu Leu Pro Ala Ala Leu Arg Arg Gly Ser Ser Gly4295 4300 4305Thr Gly Thr Ala Ala Thr Ala Ala Asn Arg Arg Gly Lys Gly Ala4310 4315 4320Glu Pro Val Ala Gly Arg Val Ala Arg Ile Val Ala Leu Leu Ala4325 4330 4335Asp Glu Arg Ser Ala Ala Leu Leu Asp Leu Val Thr Glu Gln Val4340 4345 4350Ala Glu Val Leu Gly His Ala Ser Ala Ala Glu Val Asp Pro Glu4355 4360 4365Arg Pro Phe Arg Asp Ile Gly Phe Asp Ser Leu Ala Ala Val Glu4370 4375 4380
Leu Arg Asn Arg Leu Gly Arg Leu Val Asp Leu Arg Leu Pro Thr4385 4390 4395Thr Leu Ala Phe Asp Arg Pro Thr Pro Lys Asp Val Ala Glu Trp4400 4405 4410Leu Asp Gly Glu Leu Pro Arg Pro Ala Gly Ser Ser Ala Asp Ser4415 4420 4425Ser Ala Leu Glu Gly Ile Asp Glu Leu Ala Arg Ala Val Ala Leu4430 4435 4440Leu Gly Pro Asp Asp Ala Arg Arg Ala Glu Val Arg Gln Arg Leu4445 4450 4455Thr Gly Leu Leu Ala Glu Leu Asp Thr Pro Gly His Gly Thr Ala4460 4465 4470Gly Pro Arg Asp Arg Thr Ala Pro Ala Asp Ala Glu Ser Thr Pro4475 4480 4485Ala Thr Val Ala Gly Arg Leu Asp Glu Ala Thr Asp Asp Glu Ile4490 4495 4500Phe Ala Phe Leu Asp Glu Gln Leu4505 4510<210>5<211>5835<212>DNA<213>Streptomyces mycarofaciens<220>
<221>CDS<222>(1)..(5832)<223>
<400>5atg acc gcc gag aac gac aag atc cgc agc tat ctg aag cgt gcc acc48Met Thr Ala Glu Asn Asp Lys Ile Arg Ser Tyr Leu Lys Arg Ala Thr1 5 10 15
gcc gaa ctg cac aag acc aag tcc cgc ctg gcc gag gtc gag tcg gcg96Ala Glu Leu His Lys Thr Lys Ser Arg Leu Ala Glu Val Glu Ser Ala20 25 30agc cgg gag ccg att gcg gtc gtt ggt atg gct tgt cgt tat ccg ggt144Ser Arg Glu Pro Ile Ala Val Val Gly Met Ala Cys Arg Tyr Pro Gly35 40 45ggg gtg gcg gcg ccg gag gat ttg tgg gat ctg gtg gtc gcg ggt acg192Gly Val Ala Ala Pro Glu Asp Leu Trp Asp Leu Val Val Ala Gly Thr50 55 60gac gcg atc tcc ccg ttc ccc gcc gac cgt ggc tgg gac gtc gag ggg240Asp Ala Ile Ser Pro Phe Pro Ala Asp Arg Gly Trp Asp Val Glu Gly65 70 75 80ctg tat gac ccg gac ccc gat gcg gtg ggt cgc agc tat gtg cgt gag288Leu Tyr Asp Pro Asp Pro Asp Ala Val Gly Arg Ser Tyr Val Arg Glu85 90 95ggg ggt ttt ctg cac ggg gcg gcc gag ttc gat gcg gag ttc ttc ggt336Gly Gly Phe Leu His Gly Ala Ala Glu Phe Asp Ala Glu Phe Phe Gly100 105 110gtt tcg ccg cgt gag gcg gcg gcg atg gat ccg cag cag cgg ttg ttg384Val Ser Pro Arg Glu Ala Ala Ala Met Asp Pro Gln Gln Arg Leu Leu115 120 125ttg gag acg tcg tgg gag gcg ttg gag cgg gcc ggg atc gtg ccg gct432Leu Glu Thr Ser Trp Glu Ala Leu Glu Arg Ala Gly Ile Val Pro Ala130 135 140gcg ctg cgc ggc acc cgc acc gga gtc ttc acc ggc gtg atg tat gac480Ala Leu Arg Gly Thr Arg Thr Gly Val Phe Thr Gly Val Met Tyr Asp145 150 155 160gac tac gga tcg cag ttc gat tcc gca ccg ccg gag tac gag ggc tac528Asp Tyr Gly Ser Gln Phe Asp Ser Ala Pro Pro Glu Tyr Glu Gly Tyr165 170 175ctc gtg aat ggc agc gcg ggc agc atc gca tcc ggc cgg gtt gct tac576Leu Val Asn Gly Ser Ala Gly Ser Ile Ala Ser Gly Arg Val Ala Tyr180 185 190tcc ttg ggt ttg gag ggg ccg gcg ctc acg gtg gat acc gcg tgt tcg624Ser Leu Gly Leu Glu Gly Pro Ala Leu Thr Val Asp Thr Ala Cys Ser195 200 205tcg tcg ttg gtg gcg ttg cat ctg gcg gtg cag tcg ttg cgg cgg ggt672Ser Ser Leu Val Ala Leu His Leu Ala Val Gln Ser Leu Arg Arg Gly
210 215 220gag tgc gat atg gcg ttg gcc ggt ggt gtg acg gtg atg gcg acg ccg720Glu Cys Asp Met Ala Leu Ala Gly Gly Val Thr Val Met Ala Thr Pro225 230 235 240acg gtg ttt gtg gag ttt tcc cgg cag cgt ggg ttg gct ccc gac ggg768Thr Val Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Pro Asp Gly245 250 255cgg tgc aag gcg ttt gcg gag ggt gct gat ggt act gct tgg ggt gag816Arg Cys Lys Ala Phe Ala Glu Gly Ala Asp Gly Thr Ala Trp Gly Glu260 265 270ggt gtt ggt gtg ctg ttg gtg gag cgg ctg tcc gat gcc cgt cgc ctt864Gly Val Gly Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu275 280 285ggt cac tcg gtg ttg gcg gtg gtg cgg ggg agt gcg gtt aat cag gac912Gly His Ser Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp290 295 300ggt gcc agt aat ggt ttg acg gcg ccc agt ggt ccg gct cag cag agg960Gly Ala Ser Asn Gly Leu Thr Ala Pro Ser Gly Pro Ala Gln Gln Arg305 310 315 320gtg atc cgt gag gcg ttg gcg gat gcc ggg ttg ggg tcg ggt gat gtg1008Val Ile Arg Glu Ala Leu Ala Asp Ala Gly Leu Gly Ser Gly Asp Val325 330 335gat gtg gtg gag gcg cat ggt acg ggt acg gcg ttg ggt gat ccg atc1056Asp Val Val Glu Ala His Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile340 345 350gag gct ggt gcg ttg ctg gcc acg tat ggg cgt gag cgg gtg ggt gat1104Glu Ala Gly Ala Leu Leu Ala Thr Tyr Gly Arg Glu Arg Val Gly Asp355 360 365ccg ttg tgg ttg ggg tcg ctg aag tcc aac atc ggg cac act cag gcc1152Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gln Ala370 375 380gcc gcg ggt gtg ggt ggt gtc atc aag atg gtg gag gcg ctg cgt cat1200Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Glu Ala Leu Arg His385 390 395 400ggc acg ttg cct cgc act ctc cac gtc gat gct ccc tct tcg aag gtc1248Gly Thr Leu Pro Arg Thr Leu His Val Asp Ala Pro Ser Ser Lys Val405 410 415gag tgg ggt tgg ggc gcg gtg gag ctg ttg acc gag gct cga gcc tgg1296
Glu Trp Gly Trp Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala Trp420 425 430ccc cgg cgg gcg gat cgc aag cgc cgt gcg gcc gtc tcc gcc ttc ggc1344Pro Arg Arg Ala Asp Arg Lys Arg Arg Ala Ala Val Ser Ala Phe Gly435 440 445gtc agc ggc acc aac gct cat gtc gtc atc gag gaa ccg ccc gcc gag1392Val Ser Gly Thr Asn Ala His Val Val Ile Glu Glu Pro Pro Ala Glu450 455 460gtg tcg gcc gag tcg ctg gtc gag ttg cct gct ggt gct ggt gct ggt1440Val Ser Ala Glu Ser Leu Val Glu Leu Pro Ala Gly Ala Gly Ala Gly465 470 475 480gct ggt gct ggt gct ggt gct ggg gtg tct tcg gtt gtg gcg tgg tcg1488Ala Gly Ala Gly Ala Gly Ala Gly Val Ser Ser Val Val Ala Trp Ser485 490 495ttg tcg gct cgt tcg ggt gag gcg ttg cgg gcg cag gcg gtg cgg ttg1536Leu Ser Ala Arg Ser Gly Glu Ala Leu Arg Ala Gln Ala Val Arg Leu500 505 510cgt gag cat gtg gag cgt gtt ggg gct gat ccg gtt gat gtt gcc ttt1584Arg Glu His Val Glu Arg Val Gly Ala Asp Pro Val Asp Val Ala Phe515 520 525tcg ttg gcg gtg acg cgt gcg tcg ttc ggt gag cgt gcg gtg gtc gtt1632Ser Leu Ala Val Thr Arg Ala Ser Phe Gly Glu Arg Ala Val Val Val530 535 540ggt ggt gac cgt gcg gag ttg ttg gcg ggg ctg ggg gct gtt gct gct1680Gly Gly Asp Arg Ala Glu Leu Leu Ala Gly Leu Gly Ala Val Ala Ala545 550 555 560ggg gat gcg ctg tcg ggc gtg gtg cgc ggt tcg gcg gtg cgg ggg cga1728Gly Asp Ala Leu Ser Gly Val Val Arg Gly Ser Ala Val Arg Gly Arg565 570 575aag gtt gcg gct ttg ttt acg ggt cag ggt gcg cag tgg gtt ggt atg1776Lys Val Ala Ala Leu Phe Thr Gly Gln Gly Ala Gln Trp Val Gly Met580 585 590ggg cgt gag ttg tat ggg ttg gat ggg gtg ttt gct gcg gcg ttg gat1824Gly Arg Glu Leu Tyr Gly Leu Asp Gly Val Phe Ala Ala Ala Leu Asp595 600 605gag gtt ttg ggt gtg gtg ggg gag gtg ggt ggt tgg tct ttg cgt gagl872Glu Val Leu Gly Val Val Gly Glu Val Gly Gly Trp Ser Leu Arg Glu610 615 620
gtg atg ttt ggt gag ggt ggt ggt gtt ggg gtg ggg ttg ttg gat ggt1920Val Met Phe Gly Glu Gly Gly Gly Val Gly Val Gly Leu Leu Asp Gly625 630 635 640acg gag ttt gcg cag cct gct ttg ttt gcg ttg gag gtg gcg ttg ttt1968Thr Glu Phe Ala Gln Pro Ala Leu Phe Ala Leu Glu Val Ala Leu Phe645 650 655cgg gct gtg gag gct cgg ggg gtg cgg gct tcg gtg gtg ttg ggg cat2016Arg Ala Val Glu Ala Arg Gly Val Arg Ala Ser Val Val Leu Gly His660 665 670tcg gtg ggg gag gtt gct gct gcg tgt gtg gcg ggg gtg ttt tcg ctt2064Ser Val Gly Glu Val Ala Ala Ala Cys Val Ala Gly Val Phe Ser Leu675 680 685gcg gat gcg gcg cgg ttg gtg gtg gcg cgt ggt cgg ttg atg ggt ggg2112Ala Asp Ala Ala Arg Leu Val Val Ala Arg Gly Arg Leu Met Gly Gly690 695 700ttg cct gtg ggt ggg ggg atg ttg tcg gtt cgt gcg tct gag gcc gaa2160Leu Pro Val Gly Gly Gly Met Leu Ser Val Arg Ala Ser Glu Ala Glu705 710 715 720ctt gct gat gtt gtg gct ggg ttg ggt ggt cgg gtg tcg gtg gct gcg2208Leu Ala Asp Val Val Ala Gly Leu Gly Gly Arg Val Ser Val Ala Ala725 730 735gtc aat ggt ccg gcg tcg gtg gtg ttg tct ggt gag tgt ggt gcg ttg2256Val Asn Gly Pro Ala Ser Val Val Leu Ser Gly Glu Cys Gly Ala Leu740 745 750gat gtt gtt gcg gcg cgg ttg ggt ggg cgt ggg gtg gag tgc aag cgg2304Asp Val Val Ala Ala Arg Leu Gly Gly Arg Gly Val Glu Cys Lys Arg755 760 765ttg gtg gtg tcg cat gcg ttt cat tcg gcg ttg atg gag ccg atg ttg2352Leu Val Val Ser His Ala Phe His Ser Ala Leu Met Glu Pro Met Leu770 775 780gag gag ttt cgt ggg gtt gct gag agt gtg gag tat cgg cgg ccg tgt2400Glu Glu Phe Arg Gly Val Ala Glu Ser Val Glu Tyr Arg Arg Pro Cys785 790 795 800gtg ccg gtg gtg tcg aat gtg acg ggt ggg gtg gtt ggg ttt gat gag2448Val Pro Val Val Ser Asn Val Thr Gly Gly Val Val Gly Phe Asp Glu805 810 815ttg ggt tgt gcc gag tat tgg gtg cgg cat gcg cgg gag gcg gtg cgt2496Leu Gly Cys Ala Glu Tyr Trp Val Arg His Ala Arg Glu Ala Val Arg820 825 830
ttc gct gag ggg ata cgg gct gct cgt gct ctt ggt gtg gat acg ttc2544Phe Ala Glu Gly Ile Arg Ala Ala Arg Ala Leu Gly Val Asp Thr Phe835 840 845ctg gag gtg ggt ccg cat gcg gtt ttg acg gcg atg gct ggt cag tgt2592Leu Glu Val Gly Pro His Ala Val Leu Thr Ala Met Ala Gly Gln Cys850 855 860ctt gat gga gag gag gct gac ttg gcg ttt gtg ccg gtc ctg cgg cgt2640Leu Asp Gly Glu Glu Ala Asp Leu Ala Phe Val Pro Val Leu Arg Arg865 870 875 880gat cgg ccg gca tcg cag acc ttc acc acc gca ctc gcc act ctg cac2688Asp Arg Pro Ala Ser Gln Thr Phe Thr Thr Ala Leu Ala Thr Leu His885 890 895acc cgg ggc cta ccg gta ccg ccg acg ccc tcg atg cct gcc gcc cgg2736Thr Arg Gly Leu Pro Val Pro Pro Thr Pro Ser Met Pro Ala Ala Arg900 905 910cgg atc gac ctg ccc acc tac ccc ttc caa cgg aac cgc tac tgg ctg2784Arg Ile Asp Leu Pro Thr Tyr Pro Phe Gln Arg Asn Arg Tyr Trp Leu915 920 925gcg gcc ccg ccg cgg ccc acg acc ggc ggg gtg tcg gca gcc ggt cag2832Ala Ala Pro Pro Arg Pro Thr Thr Gly Gly Val Ser Ala Ala Gly Gln930 935 940cgt gcg gtg gag cat ccg ctg ctc gcc gcc gcc gtg gaa ctc ccg ggc2880Arg Ala Val Glu His Pro Leu Leu Ala Ala Ala Val Glu Leu Pro Gly945 950 955 960gcc ggc acc gag gtg tgg acc ggc cgg atc tcc gcc gcg gac ctc ccc2928Ala Gly Thr Glu Val Trp Thr Gly Arg Ile Ser Ala Ala Asp Leu Pro965 970 975tgg ctc gcc gac cac ctg gtg tgg gac cgc gga gtg gtc ccc ggg gct2976Trp Leu Ala Asp His Leu Val Trp Asp Arg Gly Val Val Pro Gly Ala980 985 990gcc ctg ctg gag ttg gtg ctc cag gtg gga agc cgg atc gga ctg ccc 3024Ala Leu Leu Glu Leu Val Leu Gln Val Gly Ser Arg Ile Gly Leu Pro995 1000 1005cgc gtt gcc gaa ctg acc ttt gag acc gcg ctg gcc tgg gcc acg 3069Arg Val Ala Glu Leu Thr Phe Glu Thr Ala Leu Ala Trp Ala Thr1010 1015 1020gac acc ccg ctc cag atc cgg gtc gtc gtg gac gct cct gcc tcc 3114Asp Thr Pro Leu Gln Ile Arg Val Val Val Asp Ala Pro Ala Ser
1025 1030 1035gtc ccc gac ggg gcc cgt gag gtg agc ctt tac tcc cgg ccc gaa3159Val Pro Asp Gly Ala Arg Glu Val Ser Leu Tyr Ser Arg Pro Glu1040 1045 1050ccc gtc gcc cgc acc ccg cac ccc gct gga tcc ccg cac ctg gcg3204Pro Val Ala Arg Thr Pro His Pro Ala Gly Ser Pro His Leu Ala1055 1060 1065gcg gag cac ggc gac aac ggc tgg acc cgg cac gct tcc ggc gtg3249Ala Glu His Gly Asp Asn Gly Trp Thr Arg His Ala Ser Gly Val1070 1075 1080ctc gct ccg gcc gcc gac cat tcc cac gac tcc gac cca gcc gca3294Leu Ala Pro Ala Ala Asp His Ser His Asp Ser Asp Pro Ala Ala1085 1090 1095ccc agc acc ttc gcc gaa ctc acc ggt gcc tgg ccg ccc gcc ggc3339Pro Ser Thr Phe Ala Glu Leu Thr Gly Ala Trp Pro Pro Ala Gly1100 1105 1110gcc gag cct ctc gac atc gcc gag cag tac tcg ctc ttc gca gcg3384Ala Glu Pro Leu Asp Ile Ala Glu Gln Tyr Ser Leu Phe Ala Ala1115 1120 1125gtc gga gtg cgc tac gaa ggc gcc ttc cgt ggg ctg cgc gcg gcg3429Val Gly Val Arg Tyr Glu Gly Ala Phe Arg Gly Leu Arg Ala Ala1130 1135 1140tgg cgc cgc ggc gac gag atc ttc gcc gaa gtg cgg tta ccc gat3474Trp Arg Arg Gly Asp Glu Ile Phe Ala Glu Val Arg Leu Pro Asp1145 1150 1155gtg cac gcc gcc gac gcc acc cgc tac ggg gtg cat ccc gcc ctg3519Val His Ala Ala Asp Ala Thr Arg Tyr Gly Val His Pro Ala Leu1160 1165 1170ctc gac gcg gcc ctg cac ccc atc gcg ctg ctc gac ccg ttg ggc3564Leu Asp Ala Ala Leu His Pro Ile Ala Leu Leu Asp Pro Leu Gly1175 1180 1185gac ggc gga cac ggc ctg ctg ccg ttc tcc tgg acc gac gtt cag3609Asp Gly Gly His Gly Leu Leu Pro Phe Ser Trp Thr Asp Val Gln1190 1195 1200cac tac ggt tcc ggc gga cac gca ctc cgg gta cgg gtg gct gcc3654His Tyr Gly Ser Gly Gly His Ala Leu Arg Val Arg Val Ala Ala1205 1210 1215gcc gac ggc gga gcg gtg tcg atc tcc gtg gtg gac cgc gag ggt3699
Ala Asp Gly Gly Ala Val Ser Ile Ser Val Val Asp Arg Glu Gly1220 1225 1230gcc cct gtc ctc gcc gcc cgc tcc ctg gcg ctg cgc cgc atc gcc3744Ala Pro Val Leu Ala Ala Arg Ser Leu Ala Leu Arg Arg Ile Ala1235 1240 1245gcg gac cgg ctg ccc gcc gcc ccc gcc gct ccc ctg tac cgc atg3789Ala Asp Arg Leu Pro Ala Ala Pro Ala Ala Pro Leu Tyr Arg Met1250 1255 1260gac tgg ttg ccg cta ccc gag cga gtg ccc gcc gcc acg gcc gcg3834Asp Trp Leu Pro Leu Pro Glu Arg Val Pro Ala Ala Thr Ala Ala1265 1270 1275cgc tgg gcc gtc gtc ggg ccg gcg gcc gaa gtc acc gcg gcc ggg3879Arg Trp Ala Val Val Gly Pro Ala Ala Glu Val Thr Ala Ala Gly1280 1285 1290ctg cgc gcc gtc ggc gtc gat gcc cgt gcc cac gtg tcc ccc ctc3924Leu Arg Ala Val Gly Val Asp Ala Arg Ala His Val Ser Pro Leu1295 1300 1305ggc gag ccg ctg ccg ccg gag gcc ggt acg gac gcc gaa gtg tgc3969Gly Glu Pro Leu Pro Pro Glu Ala Gly Thr Asp Ala Glu Val Cys1310 1315 1320ctc ctc gac ctg acc gcg gtc gat ggc acg gcg ccc cac ggc ggg4014Leu Leu Asp Leu Thr Ala Val Asp Gly Thr Ala Pro His Gly Gly1325 1330 1335ctc ctg gac gag gtg cgc gcg acg gtg cgc cgg gcg ctg gaa gcc4059Leu Leu Asp Glu Val Arg Ala Thr Val Arg Arg Ala Leu Glu Ala1340 1345 1350gta cag acc ccg ctc gcc ggc act gat ccc ctg acg gac gcg cgt4104Val Gln Thr Pro Leu Ala Gly Thr Asp Pro Leu Thr Asp Ala Arg1355 1360 1365acg ggc act cct acc ggc ggg ccg cgg ctc gtc gtc ctc acc cgg4149Thr Gly Thr Pro Thr Gly Gly Pro Arg Leu Val Val Leu Thr Arg1370 1375 1380gga gcg gcc ggt ccg gag ggt ggc gcg gcc gat ccg gcg ggc gcc4194Gly Ala Ala Gly Pro Glu Gly Gly Ala Ala Asp Pro Ala Gly Ala1385 1390 1395gcc gtc tgg ggg ctg atc cgg gtc gcc cag acc gag cag ccc ggc4239Ala Val Trp Gly Leu Ile Arg Val Ala Gln Thr Glu Gln Pro Gly1400 1405 1410
cgc ttc acc ctg gtc gac atc gac agg gcg aag acg tcg ctg cgg4284Arg Phe Thr Leu Val Asp Ile Asp Arg Ala Lys Thr Ser Leu Arg1415 1420 1425acc ctg gcc ggg ctg ccg gcc gcg gac gcc gct cag atc gcg gtg4329Thr Leu Ala Gly Leu Pro Ala Ala Asp Ala Ala Gln Ile Ala Val1430 1435 1440cgc gac gga cgg gcc acc gtc ccc cgc ctc gta cgg gtg gtc gac4374Arg Asp Gly Arg Ala Thr Val Pro Arg Leu Val Arg Val Val Asp1445 1450 1455acc gac agc acc ggt gcc ggg gag ctg gtc gag atg ctg gac ccc4419Thr Asp Ser Thr Gly Ala Gly Glu Leu Val Glu Met Leu Asp Pro1460 1465 1470aac ggc act gtg ctg atc acc gga ggt acc gga gca ctg gcc gca4464Asn Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ala Leu Ala Ala1475 1480 1485gag acc gca cgg cac ctg gtg gaa cga cac aag gca ggt cgg ctt4509Glu Thr Ala Arg His Leu Val G1u Arg His Lys Ala Gly Arg Leu1490 1495 1500ctg ctc gtc agc agg cgc ggt gcg gag gcg ccg ggt gcc gcc gaa4554Leu Leu Val Ser Arg Arg Gly Ala Glu Ala Pro Gly Ala Ala Glu1505 1510 1515ctg gtg gcg gaa ctc gcc gcc ttg ggc gcc gag gtc acc gtc cgg4599Leu Val Ala Glu Leu Ala Ala Leu Gly Ala Glu Val Thr Val Arg1520 1525 1530gcc tgt gac gtc gct gac cgc gac gcg ctg cgc cgc ctg ctc ggt4644Ala Cys Asp Val Ala Asp Arg Asp Ala Leu Arg Arg Leu Leu Gly1535 1540 1545gag ttg ccc gcc gag cac ccc ctg agc tgt gtg gtg cac acc gcc4689Glu Leu Pro Ala Glu His Pro Leu Ser Cys Val Val His Thr Ala1550 1555 1560ggt gtg ctc gat gac ggg gtg ctc tcc gcc cag acg acc gag cgg4734Gly Val Leu Asp Asp Gly Val Leu Ser Ala Gln Thr Thr Glu Arg1565 1570 1575atc gac gcc gtg ctg cgt ccc aag gtc gac gcc gcc gtc cac ctg4779Ile Asp Ala Val Leu Arg Pro Lys Val Asp Ala Ala Val His Leu1580 1585 1590gat cag ctg acc cgt gaa ctc ggg ccg gtg cca ttg gtg ttg tac4824Asp Gln Leu Thr Arg Glu Leu Gly Pro Val Pro Leu Val Leu Tyr1595 1600 1605
tcc tcg gtc tct gcc tct ctt ggc agc gcc ggc cag gcc ggg tac4869Ser Ser Val Ser Ala Ser Leu Gly Ser Ala Gly Gln Ala Gly Tyr1610 1615 1620gcc gcg gcc aac gcg ttc ctg gac gcg ttg gcc gcc cgc cgg cgc4914Ala Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala Ala Arg Arg Arg1625 1630 1635gcc gac ggg cac cct gcg ctg tcg ctc ggc tgg ggc tgg tgg gcc4959Ala Asp Gly His Pro Ala Leu Ser Leu Gly Trp Gly Trp Trp Ala1640 1645 1650ggt gcg ggc atg gcc acc ggt ctg gag ggc gcc gac gcc gcg cgc5004Gly Ala Gly Met Ala Thr Gly Leu Glu Gly Ala Asp Ala Ala Arg1655 1660 1665atc cgg cgc tcc ggc atc gtc ccg ctc gac cct gcg gac gcg ctg5049Ile Arg Arg Ser Gly Ile Val Pro Leu Asp Pro Ala Asp Ala Leu1670 1675 1680gag ctg ctc gac cgg gcg ctg gcc cgg ccc gag ccg gcg ctg ctg5094Glu Leu Leu Asp Arg Ala Leu Ala Arg Pro Glu Pro Ala Leu Leu1685 1690 1695ccg gta cgg ctc gac ctg ccc gcc ctg cgc gct gcg gcc cgc gcc5139Pro Val Arg Leu Asp Leu Pro Ala Leu Arg Ala Ala Ala Arg Ala1700 1705 1710acc gcg cca ccg gag gtg ctg cgc gag ctc gcc ggt gtc ccg gcc5184Thr Ala Pro Pro Glu Val Leu Arg Glu Leu Ala Gly Val Pro Ala1715 1720 1725gat tcc ggg gcc gcg ctg ggt gcc ggg gga cgg gtc ggc aac ggc5229Asp Ser Gly Ala Ala Leu Gly Ala Gly Gly Arg Val Gly Asn Gly1730 1735 1740caa cgg ccc gac ccg gcc agc ccg gcc gag gca ctg gcg gcc cgg5274Gln Arg Pro Asp Pro Ala Ser Pro Ala Glu Ala Leu Ala Ala Arg1745 1750 1755ctc gcg ccg cgc tcc gca gcc gag cgc acg gcc ctc ctg ctc gac5319Leu Ala Pro Arg Ser Ala Ala Glu Arg Thr Ala Leu Leu Leu Asp1760 1765 1770ctg gtg cgt gcc gag gtc gcg gcg gtg ctg ggc cac gga gac ccc5364Leu Val Arg Ala Glu Val Ala Ala Val Leu Gly His Gly Asp Pro1775 1780 1785gcc gcg gtg ggc gcc ggc cgg tcc ttc aag gac gcc gga ttc gac5409Ala Ala Val Gly Ala Gly Arg Ser Phe Lys Asp Ala Gly Phe Asp
1790 1795 1800tcc ctc acc gcc gtc gac ctc cgc aac cgg ctg aac gcg cgc act5454Ser Leu Thr Ala Val Asp Leu Arg Asn Arg Leu Asn Ala Arg Thr1805 1810 1815ggg ctg cga ctg ccc gcg acg ctc gtg ttc gac cac ccc aca ccg5499Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp His Pro Thr Pro1820 1825 1830ttg tcc ctc gcc gag ctg ctg cgc gcc gac ctg gag gcg gcc ggc5544Leu Ser Leu Ala Glu Leu Leu Arg Ala Asp Leu Glu Ala Ala Gly1835 1840 1845ctg gtg ggg gcc acc ggt ccg gcg acg ggc gaa cca acc ggc ccc5589Leu Val Gly Ala Thr Gly Pro Ala Thr Gly Glu Pro Thr Gly Pro1850 1855 1860gag gac ctg tcc agc gtg ctg gac cgg ttg gag tcc agc ctc acc5634Glu Asp Leu Ser Ser Val Leu Asp Arg Leu Glu Ser Ser Leu Thr1865 1870 1875gcg acc gac aac ggc gac gcc cgc tcg gcc gcc gcg cgg cgg ttg5679Ala Thr Asp Asn Gly Asp Ala Arg Ser Ala Ala Ala Arg Arg Leu1880 1885 1890tgc agt ctg ctg gcc atg ctc acc gct ggc tcg ggc gag cat ccg5724Cys Ser Leu Leu Ala Met Leu Thr Ala Gly Ser Gly Glu His Pro1895 1900 1905ggg cag ggc tcc ggc gaa agc ccc cgg ggt tcc ggc gat gcg gtg5769Gly Gln Gly Ser Gly Glu Ser Pro Arg Gly Ser Gly Asp Ala Val1910 1915 1920ctc gac cgc ctc caa tcg gcc tcc gac gac gac ttg ttc gac ctt5814Leu Asp Arg Leu Gln Ser Ala Ser Asp Asp Asp Leu Phe Asp Leu1925 1930 1935ttc gac agc gat ttc cag tga 5835Phe Asp Ser Asp Phe Gln1940<210>6<211>1944<212>PRT<213>Streptomyces mycarofaciens<400>6Met Thr Ala Glu Asn Asp Lys Ile Arg Ser Tyr Leu Lys Arg Ala Thr
1 5 10 15Ala Glu Leu His Lys Thr Lys Ser Arg Leu Ala Glu Val Glu Ser Ala20 25 30Ser Arg Glu Pro Ile Ala Val Val Gly Met Ala Cys Arg Tyr Pro Gly35 40 45Gly Val Ala Ala Pro Glu Asp Leu Trp Asp Leu Val Val Ala Gly Thr50 55 60Asp Ala Ile Ser Pro Phe Pro Ala Asp Arg Gly Trp Asp Val Glu Gly65 70 75 80Leu Tyr Asp Pro Asp Pro Asp Ala Val Gly Arg Ser Tyr Val Arg Glu85 90 95Gly Gly Phe Leu His Gly Ala Ala Glu Phe Asp Ala Glu Phe Phe Gly100 105 110Val Ser Pro Arg Glu Ala Ala Ala Met Asp Pro Gln Gln Arg Leu Leu115 120 125Leu Glu Thr Ser Trp Glu Ala Leu Glu Arg Ala Gly Ile Val Pro Ala130 135 140Ala Leu Arg Gly Thr Arg Thr Gly Val Phe Thr Gly Val Met Tyr Asp145 150 155 160Asp Tyr Gly Ser Gln Phe Asp Ser Ala Pro Pro Glu Tyr Glu Gly Tyr165 170 175Leu Val Asn Gly Ser Ala Gly Ser Ile Ala Ser Gly Arg Val Ala Tyr180 185 190Ser Leu Gly Leu Glu Gly Pro Ala Leu Thr Val Asp Thr Ala Cys Ser195 200 205
Ser Ser Leu Val Ala Leu His Leu Ala Val Gln Ser Leu Arg Arg Gly210 215 220Glu Cys Asp Met Ala Leu Ala Gly Gly Val Thr Val Met Ala Thr Pro225 230 235 240Thr Val Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Pro Asp Gly245 250 255Arg Cys Lys Ala Phe Ala Glu Gly Ala Asp Gly Thr Ala Trp Gly Glu260 265 270Gly Val Gly Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu275 280 285Gly His Ser Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp290 295 300Gly Ala Ser Asn Gly Leu Thr Ala Pro Ser Gly Pro Ala Gln Gln Arg305 310 315 320Val Ile Arg Glu Ala Leu Ala Asp Ala Gly Leu Gly Ser Gly Asp Val325 330 335Asp Val Val Glu Ala His Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile340 345 350Glu Ala Gly Ala Leu Leu Ala Thr Tyr Gly Arg Glu Arg Val Gly Asp355 360 365Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gln Ala370 375 380Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Glu Ala Leu Arg His385 390 395 400Gly Thr Leu Pro Arg Thr Leu His Val Asp Ala Pro Ser Ser Lys Val405 410 415
Glu Trp Gly Trp Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala Trp420 425 430Pro Arg Arg Ala Asp Arg Lys Arg Arg Ala Ala Val Ser Ala Phe Gly435 440 445Val Ser Gly Thr Asn Ala His Val Val Ile Glu Glu Pro Pro Ala Glu450 455 460Val Ser Ala Glu Ser Leu Val Glu Leu Pro Ala Gly Ala Gly Ala Gly465 470 475 480Ala Gly Ala Gly Ala Gly Ala Gly Val Ser Ser Val Val Ala Trp Ser485 490 495Leu Ser Ala Arg Ser Gly Glu Ala Leu Arg Ala Gln Ala Val Arg Leu500 505 510Arg Glu His Val Glu Arg Val Gly Ala Asp Pro Val Asp Val Ala Phe515 520 525Ser Leu Ala Val Thr Arg Ala Ser Phe Gly Glu Arg Ala Val Val Val530 535 540Gly Gly Asp Arg Ala Glu Leu Leu Ala Gly Leu Gly Ala Val Ala Ala545 550 555 560Gly Asp Ala Leu Ser Gly Val Val Arg Gly Ser Ala Val Arg Gly Arg565 570 575Lys Val Ala Ala Leu Phe Thr Gly Gln Gly Ala Gln Trp Val Gly Met580 585 590Gly Arg Glu Leu Tyr Gly Leu Asp Gly Val Phe Ala Ala Ala Leu Asp595 600 605Glu Val Leu Gly Val Val Gly Glu Val Gly Gly Trp Ser Leu Arg Glu610 615 620
Val Met Phe Gly Glu Gly Gly Gly Val Gly Val Gly Leu Leu Asp Gly625 630 635 640Thr Glu Phe Ala Gln Pro Ala Leu Phe Ala Leu Glu Val Ala Leu Phe645 650 655Arg Ala Val Glu Ala Arg Gly Val Arg Ala Ser Val Val Leu Gly His660 665 670Ser Val Gly Glu Val Ala Ala Ala Cys Val Ala Gly Val Phe Ser Leu675 680 685Ala Asp Ala Ala Arg Leu Val Val Ala Arg Gly Arg Leu Met Gly Gly690 695 700Leu Pro Val Gly Gly Gly Met Leu Ser Val Arg Ala Ser Glu Ala Glu705 710 715 720Leu Ala Asp Val Val Ala Gly Leu Gly Gly Arg Val Ser Val Ala Ala725 730 735Val Asn Gly Pro Ala Ser Val Val Leu Ser Gly Glu Cys Gly Ala Leu740 745 750Asp Val Val Ala Ala Arg Leu Gly Gly Arg Gly Val Glu Cys Lys Arg755 760 765Leu Val Val Ser His Ala Phe His Ser Ala Leu Met Glu Pro Met Leu770 775 780Glu Glu Phe Arg Gly Val Ala Glu Ser Val Glu Tyr Arg Arg Pro Cys785 790 795 800Val Pro Val Val Ser Asn Val Thr Gly Gly Val Val Gly Phe Asp Glu805 810 815Leu Gly Cys Ala Glu Tyr Trp Val Arg His Ala Arg Glu Ala Val Arg
820 825 830Phe Ala Glu Gly Ile Arg Ala Ala Arg Ala Leu Gly Val Asp Thr Phe835 840 845Leu Glu Val Gly Pro His Ala Val Leu Thr Ala Met Ala Gly Gln Cys850 855 860Leu Asp Gly Glu Glu Ala Asp Leu Ala Phe Val Pro Val Leu Arg Arg865 870 875 880Asp Arg Pro Ala Ser Gln Thr Phe Thr Thr Ala Leu Ala Thr Leu His885 890 895Thr Arg Gly Leu Pro Val Pro Pro Thr Pro Ser Met Pro Ala Ala Arg900 905 910ArgIle Asp Leu Pro Thr Tyr Pro Phe Gln Arg Asn Arg Tyr Trp Leu915 920 925Ala Ala Pro Pro Arg Pro Thr Thr Gly Gly Val Ser Ala Ala Gly Gln930 935 940Arg Ala Val Glu His Pro Leu Leu Ala Ala Ala Val Glu Leu Pro Gly945 950 955 960Ala Gly Thr Glu Val Trp Thr Gly Arg Ile Ser Ala Ala Asp Leu Pro965 970 975Trp Leu Ala Asp His Leu Val Trp Asp Arg Gly Val Val Pro Gly Ala980 985 990Ala Leu Leu Glu Leu Val Leu Gln Val Gly Ser Arg Ile Gly Leu Pro995 1000 1005Arg Val Ala Glu Leu Thr Phe Glu Thr Ala Leu Ala Trp Ala Thr1010 1015 1020
Asp Thr Pro Leu Gln Ile Arg Val Val Val Asp Ala Pro Ala Ser1025 1030 1035Val Pro Asp Gly Ala Arg Glu Val Ser Leu Tyr Ser Arg Pro Glu1040 1045 1050Pro Val Ala Arg Thr Pro His Pro Ala Gly Ser Pro His Leu Ala1055 1060 1065Ala Glu His Gly Asp Asn Gly Trp Thr Arg His Ala Ser Gly Val1070 1075 1080Leu Ala Pro Ala Ala Asp His Ser His Asp Ser Asp Pro Ala Ala1085 1090 1095Pro Ser Thr Phe Ala Glu Leu Thr Gly Ala Trp Pro Pro Ala Gly1100 1105 1110Ala Glu Pro Leu Asp Ile Ala Glu Gln Tyr Ser Leu Phe Ala Ala1115 1120 1125Val Gly Val Arg Tyr Glu Gly Ala Phe Arg Gly Leu Arg Ala Ala1130 1135 1140Trp Arg Arg Gly Asp Glu Ile Phe Ala Glu Val Arg Leu Pro Asp1145 1150 1155Val His Ala Ala Asp Ala Thr Arg Tyr Gly Val His Pro Ala Leu1160 1165 1170Leu Asp Ala Ala Leu His Pro Ile Ala Leu Leu Asp Pro Leu Gly1175 1180 1185Asp Gly Gly His Gly Leu Leu Pro Phe Ser Trp Thr Asp Val Gln1190 1195 1200His Tyr Gly Ser Gly Gly His Ala Leu Arg Val Arg Val Ala Ala1205 1210 1215
Ala Asp Gly Gly Ala Val Ser Ile Ser Val Val Asp Arg Glu Gly1220 1225 1230Ala Pro Val Leu Ala Ala Arg Ser Leu Ala Leu Arg Arg Ile Ala1235 1240 1245Ala Asp Arg Leu Pro Ala Ala Pro Ala Ala Pro Leu Tyr Arg Met1250 1255 1260Asp Trp Leu Pro Leu Pro Glu Arg Val Pro Ala Ala Thr Ala Ala1265 1270 1275Arg Trp Ala Val Val Gly Pro Ala Ala Glu Val Thr Ala Ala Gly1280 1285 1290Leu Arg Ala Val Gly Val Asp Ala Arg Ala His Val Ser Pro Leu1295 1300 1305Gly Glu Pro Leu Pro Pro Glu Ala Gly Thr Asp Ala Glu Val Cys1310 1315 1320Leu Leu Asp Leu Thr Ala Val Asp Gly Thr Ala Pro His Gly Gly1325 1330 1335Leu Leu Asp Glu Val Arg Ala Thr Val Arg Arg Ala Leu Glu Ala1340 1345 1350Val Gln Thr Pro Leu Ala Gly Thr Asp Pro Leu Thr Asp Ala Arg1355 1360 1365Thr Gly Thr Pro Thr Gly Gly Pro Arg Leu Val Val Leu Thr Arg1370 1375 1380Gly Ala Ala Gly Pro Glu Gly Gly Ala Ala Asp Pro Ala Gly Ala1385 1390 1395Ala Val Trp Gly Leu Ile Arg Val Ala Gln Thr Glu Gln Pro Gly1400 1405 1410
Arg Phe Thr Leu Val Asp Ile Asp Arg Ala Lys Thr Ser Leu Arg1415 1420 1425Thr Leu Ala Gly Leu Pro Ala Ala Asp Ala Ala Gln Ile Ala Val1430 1435 1440Arg Asp Gly Arg Ala Thr Val Pro Arg Leu Val Arg Val Val Asp1445 1450 1455Thr Asp Ser Thr Gly Ala Gly Glu Leu Val Glu Met Leu Asp Pro1460 1465 1470Asn Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ala Leu Ala Ala1475 1480 1485Glu Thr Ala Arg His Leu Val Glu Arg His Lys Ala Gly Arg Leu1490 1495 1500Leu Leu Val Ser Arg Arg Gly Ala Glu Ala Pro Gly Ala Ala Glu1505 1510 1515Leu Val Ala Glu Leu Ala Ala Leu Gly Ala Glu Val Thr Val Arg1520 1525 1530Ala Cys Asp Val Ala Asp Arg Asp Ala Leu Arg Arg Leu Leu Gly1535 1540 1545Glu Leu Pro Ala Glu His Pro Leu Ser Cys Val Val His Thr Ala1550 1555 1560Gly Val Leu Asp Asp Gly Val Leu Ser Ala Gln Thr Thr Glu Arg1565 1570 1575Ile Asp Ala Val Leu Arg Pro Lys Val Asp Ala Ala Val His Leu1580 1585 1590Asp Gln Leu Thr Arg Glu Leu Gly Pro Val Pro Leu Val Leu Tyr
1595 1600 1605Ser Ser Val Ser Ala Ser Leu Gly Ser Ala Gly Gln Ala Gly Tyr1610 1615 1620Ala Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala Ala Arg Arg Arg1625 1630 1635Ala Asp Gly His Pro Ala Leu Ser Leu Gly Trp Gly Trp Trp Ala1640 1645 1650Gly Ala Gly Met Ala Thr Gly Leu Glu Gly Ala Asp Ala Ala Arg1655 1660 1665Ile Arg Arg Ser Gly Ile Val Pro Leu Asp Pro Ala Asp Ala Leu1670 1675 1680Glu Leu Leu Asp Arg Ala Leu Ala Arg Pro Glu Pro Ala Leu Leu1685 1690 1695Pro Val Arg Leu Asp Leu Pro Ala Leu Arg Ala Ala Ala Arg Ala1700 1705 1710Thr Ala Pro Pro Glu Val Leu Arg Glu Leu Ala Gly Val Pro Ala1715 1720 1725Asp Ser Gly Ala Ala Leu Gly Ala Gly Gly Arg Val Gly Asn Gly1730 1735 1740Gln Arg Pro Asp Pro Ala Ser Pro Ala Glu Ala Leu Ala Ala Arg1745 1750 1755Leu Ala Pro Arg Ser Ala Ala Glu Arg Thr Ala Leu Leu Leu Asp1760 1765 1770Leu Val Arg Ala Glu Val Ala Ala Val Leu Gly His Gly Asp Pro1775 1780 1785
Ala Ala Val Gly Ala Gly Arg Ser Phe Lys Asp Ala Gly Phe Asp1790 1795 1800Ser Leu Thr Ala Val Asp Leu Arg Asn Arg Leu Asn Ala Arg Thr1805 1810 1815Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp His Pro Thr Pro1820 1825 1830Leu Ser Leu Ala Glu Leu Leu Arg Ala Asp Leu Glu Ala Ala Gly1835 1840 1845Leu Val Gly Ala Thr Gly Pro Ala Thr Gly Glu Pro Thr Gly Pro1850 1855 1860Glu Asp Leu Ser Ser Val Leu Asp Arg Leu Glu Ser Ser Leu Thr1865 1870 1875Ala Thr Asp Asn Gly Asp Ala Arg Ser Ala Ala Ala Arg Arg Leu1880 1885 1890Cys Ser Leu Leu Ala Met Leu Thr Ala Gly Ser Gly Glu His Pro1895 1900 1905Gly Gln Gly Ser Gly Glu Ser Pro Arg Gly Ser Gly Asp Ala Val1910 1915 1920Leu Asp Arg Leu Gln Ser Ala Ser Asp Asp Asp Leu Phe Asp Leu1925 1930 1935Phe Asp Ser Asp Phe Gln1940<210>7<211>5847<212>DNA<213>Streptomyces mycarofaciens<220>
<221>CDS
<222>(1)..(5844)<223>
<400>7atg acc gcc gag aac gac aag atc cgc agc tat ctg aag cgt gcc acc48Met Thr Ala Glu Asn Asp Lys Ile Arg Ser Tyr Leu Lys Arg Ala Thr1 5 10 15gcc gaa ctg cac aag acc aag tcc cgc ctg gcc gag gtc gag tcg gcg96Ala Glu Leu His Lys Thr Lys Ser Arg Leu Ala Glu Val Glu Ser Ala20 25 30agc cgg gag ccg att gcg gtc gtt ggt atg gct tgt cgt tat ccg ggt144Ser Arg Glu Pro Ile Ala Val Val Gly Met Ala Cys Arg Tyr Pro Gly35 40 45ggg gtg gcg gcg ccg gag gat ttg tgg gat ctg gtg gtc gcg ggt acg192Gly Val Ala Ala Pro Glu Asp Leu Trp Asp Leu Val Val Ala Gly Thr50 55 60gac gcg atc tcc ccg ttc ccc gcc gac cgt ggc tgg gac gtc gag ggg240Asp Ala Ile Ser Pro Phe Pro Ala Asp Arg Gly Trp Asp Val Glu Gly65 70 75 80ctg tat gac ccg gac ccc gat gcg gtg ggt cgc agc tat gtg cgt gag288Leu Tyr Asp Pro Asp Pro Asp Ala Val Gly Arg Ser Tyr Val Arg Glu85 90 95ggg ggt ttt ctg cac ggg gcg gcc gag ttc gat gcg gag ttc ttc ggt336Gly Gly Phe Leu His Gly Ala Ala Glu Phe Asp Ala Glu Phe Phe Gly100 105 110gtt tcg ccg cgt gag gcg gcg gcg atg gat ccg cag cag cgg ttg ttg384Val Ser Pro Arg Glu Ala Ala Ala Met Asp Pro Gln Gln Arg Leu Leu115 120 125ttg gag acg tcg tgg gag gcg ttg gag cgg gcc ggg atc gtg ccg gct432Leu Glu Thr Ser Trp Glu Ala Leu Glu Arg Ala Gly Ile Val Pro Ala130 135 140gcg ctg cgc ggc acc cgc acc gga gtc ttc acc ggc atc tcc cag cag480Ala Leu Arg Gly Thr Arg Thr Gly Val Phe Thr Gly Ile Ser Gln Gln145 150 155 160gac tac gcc gcc cag ttg ggg gac gcg gcc gag acc tac ggc ggc cat528Asp Tyr Ala Ala Gln Leu Gly Asp Ala Ala Glu Thr Tyr Gly Gly His165 170 175gtg ctc acc gga aac ctc gga agt gtg gtc tcc ggc cgg gtt gct tac576Val Leu Thr Gly Asn Leu Gly Ser Val Val Ser Gly Arg Val Ala Tyr180 185 190
tcc ttg ggt ttg gag ggg ccg gcg ctc acg gtg gat acc gcg tgt tcg624Ser Leu Gly Leu Glu Gly Pro Ala Leu Thr Val Asp Thr Ala Cys Ser195 200 205tcg tcg ttg gtg gcg ttg cat ctg gcg gtg cag tcg ttg cgg cgg ggt672Ser Ser Leu Val Ala Leu His Leu Ala Val Gln Ser Leu Arg Arg Gly210 215 220gag tgc gat atg gcg ttg gcc ggt ggt gtg acg gtg atg gcg acg ccg720Glu Cys Asp Met Ala Leu Ala Gly Gly Val Thr Val Met Ala Thr Pro225 230 235 240acg gtg ttt gtg gag ttt tcc cgg cag cgt ggg ttg gcg tcg gat ggg768Thr Val Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Ser Asp Gly245 250 255cgg tgc aag gcg ttt gcg gag ggt gct gat ggt act gct tgg ggt gag816Arg Cys Lys Ala Phe Ala Glu Gly Ala Asp Gly Thr Ala Trp Gly Glu260 265 270ggt gtt ggt gtg ctg ttg gtg gag cgg ctg tcc gat gcc cgt cgc ctt864Gly Val Gly Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu275 280 285ggt cac tcg gtg ttg gcg gtg gtg cgg ggg agt gcg gtt aat cag gac912Gly His Ser Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp290 295 300ggt gcc agt aat ggt ttg acg gcg ccc agt ggt ccg gct cag cag agg960Gly Ala Ser Asn Gly Leu Thr Ala Pro Ser Gly Pro Ala Gln Gln Arg305 310 315 320gtg atc cgt gag gcg ttg gcg gat gcc ggg ttg ggg tcg ggt gat gtg1008Val Ile Arg Glu Ala Leu Ala Asp Ala Gly Leu Gly Ser Gly Asp Val325 330 335gat gtg gtg gag gcg cat ggt acg ggt acg gcg ttg ggt gat ccg atc1056Asp Val Val Glu Ala His Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile340 345 350gag gct ggt gcg ttg ctg gcc acg tat ggg cgt gag cgg gtg ggt gat1104Glu Ala Gly Ala Leu Leu Ala Thr Tyr Gly Arg Glu Arg Val Gly Asp355 360 365ccg ttg tgg ttg ggg tcg ctg aag tcc aac atc ggg cac act cag gcc1152Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gln Ala370 375 380gcc gcg ggt gtg ggt ggt gtc atc aag atg gtg gag gcg ctg cgt cat1200Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Glu Ala Leu Arg His
385 390 395 400ggc acg ttg cct cgc act ctc cac gtc gat gct ccc tct tcg aag gtc1248Gly Thr Leu Pro Arg Thr Leu His Val Asp Ala Pro Ser Ser Lys Val405 410 415gag tgg ggt tcg ggt gcg gtg gag ctg ttg acc gag gct cga gcc tgg1296Glu Trp Gly Ser Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala Trp420 425 430ccc cgg cgg gcg gat cgc aag cgc cgt gcg gcc gtc tcc gcc ttc ggc1344Pro Arg Arg Ala Asp Arg Lys Arg Arg Ala Ala Val Ser Ala Phe Gly435 440 445gtc agc ggc acc aac gct cat gtc gtc atc gag gaa ccg ccc gcc gag1392Val Ser Gly Thr Asn Ala His Val Val Ile Glu Glu Pro Pro Ala Glu450 455 460gtg tcg gcc gag tcg ctg gtc gag ttg cct gct ggt gct ggt gct ggt1440Val Ser Ala Glu Ser Leu Val Glu Leu Pro Ala Gly Ala Gly Ala Gly465 470 475 480gct ggt gct ggt gct ggt gct ggt gct ggt gct ggg gtg tct tcg gtt1488Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Val Ser Ser Val485 490 495gtg gcg tgg tcg ttg tcg gct cgt tcg ggt gag gcg ttg cgg gcg cag1536Val Ala Trp Ser Leu Ser Ala Arg Ser Gly Glu Ala Leu Arg Ala Gln500 505 510gcg gtg cgg ttg cgt gag cat gtg gag cgt gtt ggg gct gat ccg gtt1584Ala Val Arg Leu Arg Glu His Val Glu Arg Val Gly Ala Asp Pro Val515 520 525gat gtt gcc ttt tcg ttg gcg gtg acg cgt gcg tcg ttc ggt gag cgt1632Asp Val Ala Phe Ser Leu Ala Val Thr Arg Ala Ser Phe Gly Glu Arg530 535 540gcg gtg gtc gtt ggt ggt gac cgt gcg gag ttg ttg gcg ggg ctg gggl680Ala Val Val Val Gly Gly Asp Arg Ala Glu Leu Leu Ala Gly Leu Gly545 550 555 560gct gtt gct gct ggg gat gcg ctg tcg ggc gtg gtg cgt ggt tcg gcg1728Ala Val Ala Ala Gly Asp Ala Leu Ser Gly Val Val Arg Gly Ser Ala565 570 575gtg cgg ggg cga aag gtt gcg gct ttg ttt acg ggt cag ggt gcg cag1776Val Arg Gly Arg Lys Val Ala Ala Leu Phe Thr Gly Gln Gly Ala Gln580 585 590tgg gtt ggt atg ggg cgt gag ttg tat ggg ttg gat ggg gtg ttt gct1824
Trp Val Gly Met Gly Arg Glu Leu Tyr Gly Leu Asp Gly Val Phe Ala595 600 605gcg gcg ttg gat gag gtt ttg ggt gtg gtg ggg gag gtg ggt ggt tgg1872Ala Ala Leu Asp Glu Val Leu Gly Val Val Gly Glu Val Gly Gly Trp610 615 620tct ttg cgt gag gtg atg ttt ggt gag ggt ggt ggt gtt ggg gtg ggg1920Ser Leu Arg Glu Val Met Phe Gly Glu Gly Gly Gly Val Gly Val Gly625 630 635 640ttg ttg gat ggt acg gag ttt gcg cag cct gct ttg ttt gcg ttg gag1968Leu Leu Asp Gly Thr Glu Phe Ala Gln Pro Ala Leu Phe Ala Leu Glu645 650 655gtg gcg ttg ttt cgg gct gtg gag gct cgg ggg gtg cgg gct tcg gtg2016Val Ala Leu Phe Arg Ala Val Glu Ala Arg Gly Val Arg Ala Ser Val660 665 670gtg ttg ggg cat tcg gtg ggg gag gtt gct gct gcg tgt gtg gcg ggg2064Val Leu Gly His Ser Val Gly Glu Val Ala Ala Ala Cys Val Ala Gly675 680 685gtg ttt tcg ctt gcg gat gcg gcg cgg ttg gtg gtg gcg cgt ggt cgg2112Val Phe Ser Leu Ala Asp Ala Ala Arg Leu Val Val Ala Arg Gly Arg690 695 700ttg atg ggt ggg ttg cct gtg ggt ggg ggg atg ttg tcg gtt cgt gcg2160Leu Met Gly Gly Leu Pro Val Gly Gly Gly Met Leu Ser Val Arg Ala705 710 715 720tct gag gcc gaa ctt gct gat gtt gtg gct ggg ttg ggt ggt cgg gtg2208Ser Glu Ala Glu Leu Ala Asp Val Val Ala Gly Leu Gly Gly Arg Val725 730 735tcg gtg gct gcg gtc aat ggt ccg gcg tcg gtg gtg ttg tct ggt gag2256Ser Val Ala Ala Val Asn Gly Pro Ala Ser Val Val Leu Ser Gly Glu740 745 750tgt ggt gcg ttg gat gtt gtt gcg gcg cgg ttg ggt ggg cgt ggg gtg2304Cys Gly Ala Leu Asp Val Val Ala Ala Arg Leu Gly Gly Arg Gly Val755 760 765gag tgc aag cgg ttg gtg gtg tcg cat gcg ttt cat tcg gcg ttg atg2352Glu Cys Lys Arg Leu Val Val Ser His Ala Phe His Ser Ala Leu Met770 775 780gag ccg atg ttg gag gag ttt cgt ggg gtt gct gag agt gtg gag tat2400Glu Pro Met Leu Glu Glu Phe Arg Gly Val Ala Glu Ser Val Glu Tyr785 790 795 800
cgg cgg ccg tgt gtg ccg gtg gtg tcg aat gtg acg ggt ggg gtg gtt2448Arg Arg Pro Cys Val Pro Val Val Ser Asn Val Thr Gly Gly Val Val805 810 815ggg ttt gat gag ttg ggt tgt gcc gag tat tgg gtg cgg cat gcg cgg2496Gly Phe Asp Glu Leu Gly Cys Ala Glu Tyr Trp Val Arg His Ala Arg820 825 830gag gcg gtg cgt ttc gct gag ggg ata cgg gct gct cgt gct ctt ggt2544Glu Ala Val Arg Phe Ala Glu Gly Ile Arg Ala Ala Arg Ala Leu Gly835 840 845gtg gat acg ttc ctg gag gtg ggt ccg cat gcg gtt ttg acg gcg atg2592Val Asp Thr Phe Leu Glu Val Gly Pro His Ala Val Leu Thr Ala Met850 855 860gct ggt cag tgt ctt gat gga gag gag gct gac ttg gcg ttt gtg ccg2640Ala Gly Gln Cys Leu Asp Gly Glu Glu Ala Asp Leu Ala Phe Val Pro865 870 875 880gtc ctg cgg cgt gat cgg ccg gca tcg cag acc ttc acc acc gca ctc2688Val Leu Arg Arg Asp Arg Pro Ala Ser Gln Thr Phe Thr Thr Ala Leu885 890 895gcc act ctg cac acc cgg ggc cta ccg gta ccg ccg acg ccc tcg atg2736Ala Thr Leu His Thr Arg Gly Leu Pro Val Pro Pro Thr Pro Ser Met900 905 910cct gcc gcc cgg cgg atc gac ctg ccc acc tac ccc ttc caa cgg aac2784Pro Ala Ala Arg Arg Ile Asp Leu Pro Thr Tyr Pro Phe Gln Arg Asn915 920 925cgc tac tgg ctg gcg gcc ccg ccg cgg ccc acg acc ggc ggg gtg tcg2832Arg Tyr Trp Leu Ala Ala Pro Pro Arg Pro Thr Thr Gly Gly Val Ser930 935 940gca gcc ggt cag cgt gcg gtg gag cat ccg ctg ctc gcc gcc gcc gtg2880Ala Ala Gly Gln Arg Ala Val Glu His Pro Leu Leu Ala Ala Ala Val945 950 955 960gaa ctc ccg ggc gcc ggc acc gag gtg tgg acc ggc cgg atc tcc gcc2928Glu Leu Pro Gly Ala Gly Thr Glu Val Trp Thr Gly Arg Ile Ser Ala965 970 975gcg gac ctc ccc tgg ctc gcc gac cac ctg gtg tgg gac cgc gga gtg2976Ala Asp Leu Pro Trp Leu Ala Asp His Leu Val Trp Asp Arg Gly Val980 985 990gtc ccc ggg gct gcc ctg ctg gag ttg gtg ctc cag gtg gga agc cgg 3024Val Pro Gly Ala Ala Leu Leu Glu Leu Val Leu Gln Val Gly Ser Arg995 1000 1005
atc gga ctg ccc cgc gtt gcc gaa ctg acc ttt gag acc gcg ctg3069Ile Gly Leu Pro Arg Val Ala Glu Leu Thr Phe Glu Thr Ala Leu1010 1015 1020gcc tgg gcc acg gac acc ccg ctc cag atc cgg gtc gtc gtg gac3114Ala Trp Ala Thr Asp Thr Pro Leu Gln Ile Arg Val Val Val Asp1025 1030 1035gct cct gcc tcc gtc ccc gac ggg gcc cgt gag gtg agc ctt tac3159Ala Pro Ala Ser Val Pro Asp Gly Ala Arg Glu Val Ser Leu Tyr1040 1045 1050tcc cgg ccc gaa ccc gtc gcc cgc acc ccg cac ccc gct gga tcc3204Ser Arg Pro Glu Pro Val Ala Arg Thr Pro His Pro Ala Gly Ser1055 1060 1065ccg cac ctg gcg gcg gag cac ggc gac aac ggc tgg acc cgg cac3249Pro His Leu Ala Ala Glu His Gly Asp Asn Gly Trp Thr Arg His1070 1075 1080gct tcc ggc gtg ctc gct ccg gcc gcc gac cat tcc cac gac tcc3294Ala Ser Gly Val Leu Ala Pro Ala Ala Asp His Ser His Asp Ser1085 1090 1095gac cca gcc gca ccc agc acc ttc gcc gaa ctc acc ggt gcc tgg3339Asp Pro Ala Ala Pro Ser Thr Phe Ala Glu Leu Thr Gly Ala Trp1100 1105 1110ccg ccc gcc ggc gcc gag cct ctc gac atc gcc gag cag tac tcg3384Pro Pro Ala Gly Ala Glu Pro Leu Asp Ile Ala Glu Gln Tyr Ser1115 1120 1125ctc ttc gca gcg gtc gga gtg cgc tac gaa ggc gcc ttc cgt ggg3429Leu Phe Ala Ala Val Gly Val Arg Tyr Glu Gly Ala Phe Arg Gly1130 1135 1140ctg cgc gcg gcg tgg cgc cgc ggc gac gag atc ttc gcc gaa gtg3474Leu Arg Ala Ala Trp Arg Arg Gly Asp Glu Ile Phe Ala Glu Val1145 1150 1155cgg tta ccc gat gtg cac gcc gcc gac gcc acc cgc tac ggg gtg3519Arg Leu Pro Asp Val His Ala Ala Asp Ala Thr Arg Tyr Gly Val1160 1165 1170cat ccc gcc ctg ctc gac gcg gcc ctg cac ccc atc gcg ctg ctc3564His Pro Ala Leu Leu Asp Ala Ala Leu His Pro Ile Ala Leu Leu1175 1180 1185gac ccg ttg ggc gac ggc gga cac ggc ctg ctg ccg ttc tcc tgg3609Asp Pro Leu Gly Asp Gly Gly His Gly Leu Leu Pro Phe Ser Trp
1190 1195 1200acc gac gtt cag cac tac ggt tcc ggc gga cac gca ctc cgg gta3654Thr Asp Val Gln His Tyr Gly Ser Gly Gly His Ala Leu Arg Val1205 1210 1215cgg gtg gct gcc gcc gac ggc gga gcg gtg tcg atc tcc gtg gtg3699Arg Val Ala Ala Ala Asp Gly Gly Ala Val Ser Ile Ser Val Val1220 1225 1230gac cgc gag ggt gcc cct gtc ctc gcc gcc cgc tcc ctg gcg ctg3744Asp Arg Glu Gly Ala Pro Val Leu Ala Ala Arg Ser Leu Ala Leu1235 1240 1245cgc cgc atc gcc gcg gac cgg ctg ccc gcc gcc ccc gcc gct ccc3789Arg Arg Ile Ala Ala Asp Arg Leu Pro Ala Ala Pro Ala Ala Pro1250 1255 1260ctg tac cgc atg gac tgg ttg ccg cta ccc gag cga gtg ccc gcc3834Leu Tyr Arg Met Asp Trp Leu Pro Leu Pro Glu Arg Val Pro Ala1265 1270 1275gcc acg gcc gcg cgc tgg gcc gtc gtc ggg ccg gcg gcc gaa gtc3879Ala Thr Ala Ala Arg Trp Ala Val Val Gly Pro Ala Ala Glu Val1280 1285 1290acc gcg gcc ggg ctg cgc gcc gtc ggc gtc gat gcc cgt gcc cac3924Thr Ala Ala Gly Leu Arg Ala Val Gly Val Asp Ala Arg Ala His1295 1300 1305gtg tcc ccc ctc ggc gag ccg ctg ccg ccg gag gcc ggt acg gac3969Val Ser Pro Leu Gly Glu Pro Leu Pro Pro Glu Ala Gly Thr Asp1310 1315 1320gcc gaa gtg tgc ctc ctc gac ctg acc gcg gtc gat ggc acg gcg4014Ala Glu Val Cys Leu Leu Asp Leu Thr Ala Val Asp Gly Thr Ala1325 1330 1335ccc cac ggc ggg ctc ctg gac gag gtg cgc gcg acg gtg cgc cgg4059Pro His Gly Gly Leu Leu Asp Glu Val Arg Ala Thr Val Arg Arg1340 1345 1350gcg ctg gaa gcc gta cag acc ccg ctc gcc ggc act gat ccc ctg4104Ala Leu Glu Ala Val Gln Thr Pro Leu Ala Gly Thr Asp Pro Leu1355 1360 1365acg gac gcg cgt acg ggc act cct acc ggc ggg ccg cgg ctc gtc4149Thr Asp Ala Arg Thr Gly Thr Pro Thr Gly Gly Pro Arg Leu Val1370 1375 1380gtc ctc acc cgg gga gcg gcc ggt ccg gag ggt ggc gcg gcc gat4194
Val Leu Thr Arg Gly Ala Ala Gly Pro Glu Gly Gly Ala Ala Asp1385 1390 1395ccg gcg ggc gcc gcc gtc tgg ggg ctg atc cgg gtc gcc cag acc4239Pro Ala Gly Ala Ala Val Trp Gly Leu Ile Arg Val Ala Gln Thr1400 1405 1410gag cag ccc ggc cgc ttc acc ctg gtc gac atc gac agg gcg aag4284Glu Gln Pro Gly Arg Phe Thr Leu Val Asp Ile Asp Arg Ala Lys1415 1420 1425acg tcg ctg cgg acc ctg gcc ggg ctg ccg gcc gcg gac gcc gct4329Thr Ser Leu Arg Thr Leu Ala Gly Leu Pro Ala Ala Asp Ala Ala1430 1435 1440cag atc gcg gtg cgc gac gga cgg gcc acc gtc ccc cgc ctc gta4374Gln Ile Ala Val Arg Asp Gly Arg Ala Thr Val Pro Arg Leu Val1445 1450 1455cgg gtg gtc gac acc gac agc acc ggt gcc ggg gag ctg gtc gag4419Arg Val Val Asp Thr Asp Ser Thr Gly Ala Gly Glu Leu Val Glu1460 1465 1470atg ctg gac ccc aac ggc act gtg ctg atc acc gga ggt acc gga4464Met Leu Asp Pro Asn Gly Thr Val Leu Ile Thr Gly Gly Thr Gly1475 1480 1485gca ctg gcc gca gag acc gca cgg cac ctg gtg gaa cga cac aag4509Ala Leu Ala Ala Glu Thr Ala Arg His Leu Val Glu Arg His Lys1490 1495 1500gca ggt cgg ctt ctg ctc gtc agc agg cgc ggt gcg gag gcg ccg4554Ala Gly Arg Leu Leu Leu Val Ser Arg Arg Gly Ala Glu Ala Pro1505 1510 1515ggt gcc gcc gaa ctg gtg gcg gaa ctc gcc gcc ttg ggc gcc gag4599Gly Ala Ala Glu Leu Val Ala Glu Leu Ala Ala Leu Gly Ala Glu1520 1525 1530gtc acc gtc cgg gcc tgt gac gtc gct gac cgc gac gcg ctg cgc4644Val Thr Val Arg Ala Cys Asp Val Ala Asp Arg Asp Ala Leu Arg1535 1540 1545cgc ctg ctc ggt gag ttg ccc gcc gag cac ccc ctg agc tgt gtg4689Arg Leu Leu Gly Glu Leu Pro Ala Glu His Pro Leu Ser Cys Val1550 1555 1560gtg cac acc gcc ggt gtg ctc gat gac ggg gtg ctc tcc gcc cag4734Val His Thr Ala Gly Val Leu Asp Asp Gly Val Leu Ser Ala Gln1565 1570 1575
acg acc gag cgg atc gac gcc gtg ctg cgt ccc aag gtc gac gcc4779Thr Thr Glu Arg Ile Asp Ala Val Leu Arg Pro Lys Val Asp Ala1580 1585 1590gcc gtc cac ctg gat cag ctg acc cgt gaa ctc ggg ccg gtg cca4824Ala Val His Leu Asp Gln Leu Thr Arg Glu Leu Gly Pro Val Pro1595 1600 1605ttg gtg ttg tac tcc tcg gtc tct gcc tct ctt ggc agc gcc ggc4869Leu Val Leu Tyr Ser Ser Val Ser Ala Ser Leu Gly Ser Ala Gly1610 1615 1620cag gcc ggg tac gcc gcg gcc aac gcg ttc ctg gac gcg ttg gcc4914Gln Ala Gly Tyr Ala Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala1625 1630 1635gcc cgc cgg cgc gcc gac ggg cac cct gcg ctg tcg ctc ggc tgg4959Ala Arg Arg Arg Ala Asp Gly His Pro Ala Leu Ser Leu Gly Trp1640 1645 1650ggc tgg tgg gcc ggt gcg ggc atg gcc acc ggt ctg gag ggc gcc5004Gly Trp Trp Ala Gly Ala Gly Met Ala Thr Gly Leu Glu Gly Ala1655 1660 1665gac gcc gcg cgc atc cgg cgc tcc ggc atc gtc ccg ctc gac cct5049Asp Ala Ala Arg Ile Arg Arg Ser Gly Ile Val Pro Leu Asp Pro1670 1675 1680gcg gac gcg ctg gag ctg ctc gac cgg gcg ctg gcc cgg ccc gag5094Ala Asp Ala Leu Glu Leu Leu Asp Arg Ala Leu Ala Arg Pro Glu1685 1690 1695ccg gcg ctg ctg ccg gta cgg ctc gac ctg ccc gcc ctg cgc gct5139Pro Ala Leu Leu Pro Val Arg Leu Asp Leu Pro Ala Leu Arg Ala1700 1705 1710gcg gcc cgc gcc acc gcg cca ccg gag gtg ctg cgc gag ctc gcc5184Ala Ala Arg Ala Thr Ala Pro Pro Glu Val Leu Arg Glu Leu Ala1715 1720 1725ggt gtc ccg gcc gat tcc ggg gcc gcg ctg ggt gcc ggg gga cgg5229Gly Val Pro Ala Asp Ser Gly Ala Ala Leu Gly Ala Gly Gly Arg1730 1735 1740gtc ggc aac ggc caa cgg ccc gac ccg gcc agc ccg gcc gag gca5274Val Gly Asn Gly Gln Arg Pro Asp Pro Ala Ser Pro Ala Glu Ala1745 1750 1755ctg gcg gcc cgg ctc gcg ccg cgc tcc gca gcc gag cgc acg gcc5319Leu Ala Ala Arg Leu Ala Pro Arg Ser Ala Ala Glu Arg Thr Ala1760 1765 1770
ctc ctg ctc gac ctg gtg cgt gcc gag gtc gcg gcg gtg ctg ggc5364Leu Leu Leu Asp Leu Val Arg Ala Glu Val Ala Ala Val Leu Gly1775 1780 1785cac gga gac ccc gcc gcg gtg ggc gcc ggc cgg tcc ttc aag gac5409His Gly Asp Pro Ala Ala Val Gly Ala Gly Arg Ser Phe Lys Asp1790 1795 1800gcc gga ttc gac tcc ctc acc gcc gtc gac ctc cgc aac cgg ctg5454Ala Gly Phe Asp Ser Leu Thr Ala Val Asp Leu Arg Asn Arg Leu1805 1810 1815aac gcg cgc act ggg ctg cga ctg ccc gcg acg ctc gtg ttc gac5499Asn Ala Arg Thr Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp1820 1825 1830cac ccc aca ccg ttg tcc ctc gcc gag ctg ctg cgc gcc gac ctg5544His Pro Thr Pro Leu Ser Leu Ala Glu Leu Leu Arg Ala Asp Leu1835 1840 1845gag gcg gcc ggc ctg gtg ggg gcc acc ggt ccg gcg acg ggc gaa5589Glu Ala Ala Gly Leu Val Gly Ala Thr Gly Pro Ala Thr Gly Glu1850 1855 1860cca acc ggc ccc gag gac ctg tcc agc gtg ctg gac cgg ttg gag5634Pro Thr Gly Pro Glu Asp Leu Ser Ser Val Leu Asp Arg Leu Glu1865 1870 1875tcc agc ctc acc gcg acc gac aac ggc gac gcc cgc tcg gcc gcc5679Ser Ser Leu Thr Ala Thr Asp Asn Gly Asp Ala Arg Ser Ala Ala1880 1885 1890gcg cgg cgg ttg tgc agt ctg ctg gcc atg ctc acc gct ggc tcg5724Ala Arg Arg Leu Cys Ser Leu Leu Ala Met Leu Thr Ala Gly Ser1895 1900 1905ggc gag cat ccg ggg cag ggc tcc ggc gaa agc ccc cgg ggt tcc5769Gly Glu His Pro Gly Gln Gly Ser Gly Glu Ser Pro Arg Gly Ser1910 1915 1920ggc gat gcg gtg ctc gac cgc ctc caa tcg gcc tcc gac gac gac5814Gly Asp Ala Val Leu Asp Arg Leu Gln Ser Ala Ser Asp Asp Asp1925 1930 1935ttg ttc gac ctt ttc gac agc gat ttc cag tga 5847Leu Phe Asp Leu Phe Asp Ser Asp Phe Gln1940 1945<210>8
<211>1948<212>PRT<213>Streptomyces mycarofaciens<400>8Met Thr Ala Glu Asn Asp Lys Ile Arg Ser Tyr Leu Lys Arg Ala Thr1 5 10 15Ala Glu Leu His Lys Thr Lys Ser Arg Leu Ala Glu Val Glu Ser Ala20 25 30Ser Arg Glu Pro Ile Ala Val Val Gly Met Ala Cys Arg Tyr Pro Gly35 40 45Gly Val Ala Ala Pro Glu Asp Leu Trp Asp Leu Val Val Ala Gly Thr50 55 60Asp Ala Ile Ser Pro Phe Pro Ala Asp Arg Gly Trp Asp Val Glu Gly65 70 75 80Leu Tyr Asp Pro Asp Pro Asp Ala Val Gly Arg Ser Tyr Val Arg Glu85 90 95Gly Gly Phe Leu His Gly Ala Ala Glu Phe Asp Ala Glu Phe Phe Gly100 105 110Val Ser Pro Arg Glu Ala Ala Ala Met Asp Pro Gln Gln Arg Leu Leu115 120 125Leu Glu Thr Ser Trp Glu Ala Leu Glu Arg Ala Gly Ile Val Pro Ala130 135 140Ala Leu Arg Gly Thr Arg Thr Gly Val Phe Thr Gly Ile Ser Gln Gln145 150 155 160Asp Tyr Ala Ala Gln Leu Gly Asp Ala Ala Glu Thr Tyr Gly Gly His165 170 175Val Leu Thr Gly Asn Leu Gly Ser Val Val Ser Gly Arg Val Ala Tyr
180 185 190Ser Leu Gly Leu Glu Gly Pro Ala Leu Thr Val Asp Thr Ala Cys Ser195 200 205Ser Ser Leu Val Ala Leu His Leu Ala Val Gln Ser Leu Arg Arg Gly210 215 220Glu Cys Asp Met Ala Leu Ala Gly Gly Val Thr Val Met Ala Thr Pro225 230 235 240Thr Val Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Ser Asp Gly245 250 255Arg Cys Lys Ala Phe Ala Glu Gly Ala Asp Gly Thr Ala Trp Gly Glu260 265 270Gly Val Gly Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu275 280 285Gly His Ser Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp290 295 300Gly Ala Ser Asn Gly Leu Thr Ala Pro Ser Gly Pro Ala Gln Gln Arg305 310 315 320Val Ile Arg Glu Ala Leu Ala Asp Ala Gly Leu Gly Ser Gly Asp Val325 330 335Asp Val Val Glu Ala His Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile340 345 350Glu Ala Gly Ala Leu Leu Ala Thr Tyr Gly Arg Glu Arg Val Gly Asp355 360 365Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile Gly His Thr Gln Ala370 375 380
Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Glu Ala Leu Arg His385 390 395 400Gly Thr Leu Pro Arg Thr Leu His Val Asp Ala Pro Ser Ser Lys Val405 410 415Glu Trp Gly Ser Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala Trp420 425 430Pro Arg Arg Ala Asp Arg Lys Arg Arg Ala Ala Val Ser Ala Phe Gly435 440 445Val Ser Gly Thr Asn Ala His Val Val Ile Glu Glu Pro Pro Ala Glu450 455 460Val Ser Ala Glu Ser Leu Val Glu Leu Pro Ala Gly Ala Gly Ala Gly465 470 475 480Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Ala Gly Val Ser Ser Val485 490 495Val Ala Trp Ser Leu Ser Ala Arg Ser Gly Glu Ala Leu Arg Ala Gln500 505 510Ala Val Arg Leu Arg Glu His Val Glu Arg Val Gly Ala Asp Pro Val515 520 525Asp Val Ala Phe Ser Leu Ala Val Thr Arg Ala Ser Phe Gly Glu Arg530 535 540Ala Val Val Val Gly Gly Asp Arg Ala Glu Leu Leu Ala Gly Leu Gly545 550 555 560Ala Val Ala Ala Gly Asp Ala Leu Ser Gly Val Val Arg Gly Ser Ala565 570 575Val Arg Gly Arg Lys Val Ala Ala Leu Phe Thr Gly Gln Gly Ala Gln580 585 590
Trp Val Gly Met Gly Arg Glu Leu Tyr Gly Leu Asp Gly Val Phe Ala595 600 605Ala Ala Leu Asp Glu Val Leu Gly Val Val Gly Glu Val Gly Gly Trp610 615 620Ser Leu Arg Glu Val Met Phe Gly Glu Gly Gly Gly Val Gly Val Gly625 630 635 640Leu Leu Asp Gly Thr Glu Phe Ala Gln Pro Ala Leu Phe Ala Leu Glu645 650 655Val Ala Leu Phe Arg Ala Val Glu Ala Arg Gly Val Arg Ala Ser Val660 665 670Val Leu Gly His Ser Val Gly Glu Val Ala Ala Ala Cys Val Ala Gly675 680 685Val Phe Ser Leu Ala Asp Ala Ala Arg Leu Val Val Ala Arg Gly Arg690 695 700Leu Met Gly Gly Leu Pro Val Gly Gly Gly Met Leu Ser Val Arg Ala705 710 715 720Ser Glu Ala Glu Leu Ala Asp Val Val Ala Gly Leu Gly Gly Arg Val725 730 735Ser Val Ala Ala Val Asn Gly Pro Ala Ser Val Val Leu Ser Gly Glu740 745 750Cys Gly Ala Leu Asp Val Val Ala Ala Arg Leu Gly Gly Arg Gly Val755 760 765Glu Cys Lys Arg Leu Val Val Ser His Ala Phe His Ser Ala Leu Met770 775 780Glu Pro Met Leu Glu Glu Phe Arg Gly Val Ala Glu Ser Val Glu Tyr785 790 795 800
Arg Arg Pro Cys Val Pro Val Val Ser Asn Val Thr Gly Gly Val Val805 810 815Gly Phe Asp Glu Leu Gly Cys Ala Glu Tyr Trp Val Arg His Ala Arg820 825 830Glu Ala Val Arg Phe Ala Glu Gly Ile Arg Ala Ala Arg Ala Leu Gly835 840 845Val Asp Thr Phe Leu Glu Val Gly Pro His Ala Val Leu Thr Ala Met850 855 860Ala Gly Gln Cys Leu Asp Gly Glu Glu Ala Asp Leu Ala Phe Val Pro865 870 875 880Val Leu Arg Arg Asp Arg Pro Ala Ser Gln Thr Phe Thr Thr Ala Leu885 890 895Ala Thr Leu His Thr Arg Gly Leu Pro Val Pro Pro Thr Pro Ser Met900 905 910Pro Ala Ala Arg Arg Ile Asp Leu Pro Thr Tyr Pro Phe Gln Arg Asn915 920 925Arg Tyr Trp Leu Ala Ala Pro Pro Arg Pro Thr Thr Gly Gly Val Ser930 935 940Ala Ala Gly Gln Arg Ala Val Glu His Pro Leu Leu Ala Ala Ala Val945 950 955 960Glu Leu Pro Gly Ala Gly Thr Glu Val Trp Thr Gly Arg Ile Ser Ala965 970 975Ala Asp Leu Pro Trp Leu Ala Asp His Leu Val Trp Asp Arg Gly Val980 985 990Val Pro Gly Ala Ala Leu Leu Glu Leu Val Leu Gln Val Gly Ser Arg
995 1000 1005Ile Gly Leu Pro Arg Val Ala Glu Leu Thr Phe Glu Thr Ala Leu1010 1015 1020Ala Trp Ala Thr Asp Thr Pro Leu Gln Ile Arg Val Val Val Asp1025 1030 1035Ala Pro Ala Ser Val Pro Asp Gly Ala Arg Glu Val Ser Leu Tyr1040 1045 1050Ser Arg Pro Glu Pro Val Ala Arg Thr Pro His Pro Ala Gly Ser1055 1060 1065Pro His Leu Ala Ala Glu His Gly Asp Asn Gly Trp Thr Arg His1070 1075 1080Ala Ser Gly Val Leu Ala Pro Ala Ala Asp His Ser His Asp Ser1085 1090 1095Asp Pro Ala Ala Pro Ser Thr Phe Ala Glu Leu Thr Gly Ala Trp1100 1105 1110Pro Pro Ala Gly Ala Glu Pro Leu Asp Ile Ala Glu Gln Tyr Ser1115 1120 1125Leu Phe Ala Ala Val Gly Val Arg Tyr Glu Gly Ala Phe Arg Gly1130 1135 1140Leu Arg Ala Ala Trp Arg Arg Gly Asp Glu Ile Phe Ala Glu Val1145 1150 1155Arg Leu Pro Asp Val His Ala Ala Asp Ala Thr Arg Tyr Gly Val1160 1165 1170His Pro Ala Leu Leu Asp Ala Ala Leu His Pro Ile Ala Leu Leu1175 1180 1185
Asp Pro Leu Gly Asp Gly Gly His Gly Leu Leu Pro Phe Ser Trp1190 1195 1200Thr Asp Val Gln His Tyr Gly Ser Gly Gly His Ala Leu Arg Val1205 1210 1215Arg Val Ala Ala Ala Asp Gly Gly Ala Val Ser Ile Ser Val Val1220 1225 1230Asp Arg Glu Gly Ala Pro Val Leu Ala Ala Arg Ser Leu Ala Leu1235 1240 1245Arg Arg Ile Ala Ala Asp Arg Leu Pro Ala Ala Pro Ala Ala Pro1250 1255 1260Leu Tyr Arg Met Asp Trp Leu Pro Leu Pro Glu Arg Val Pro Ala1265 1270 1275Ala Thr Ala Ala Arg Trp Ala Val Val Gly Pro Ala Ala Glu Val1280 1285 1290Thr Ala Ala Gly Leu Arg Ala Val Gly Val Asp Ala Arg Ala His1295 1300 1305Val Ser Pro Leu Gly Glu Pro Leu Pro Pro Glu Ala Gly Thr Asp1310 1315 1320Ala Glu Val Cys Leu Leu Asp Leu Thr Ala Val Asp Gly Thr Ala1325 1330 1335Pro His Gly Gly Leu Leu Asp Glu Val Arg Ala Thr Val Arg Arg1340 1345 1350Ala Leu Glu Ala Val Gln Thr Pro Leu Ala Gly Thr Asp Pro Leu1355 1360 1365Thr Asp Ala Arg Thr Gly Thr Pro Thr Gly Gly Pro Arg Leu Val1370 1375 1380
Val Leu Thr Arg Gly Ala Ala Gly Pro Glu Gly Gly Ala Ala Asp1385 1390 1395Pro Ala Gly Ala Ala Val Trp Gly Leu Ile Arg Val Ala Gln Thr1400 1405 1410Glu Gln Pro Gly Arg Phe Thr Leu Val Asp Ile Asp Arg Ala Lys1415 1420 1425Thr Ser Leu Arg Thr Leu Ala Gly Leu Pro Ala Ala Asp Ala Ala1430 1435 1440Gln Ile Ala Val Arg Asp Gly Arg Ala Thr Val Pro Arg Leu Val1445 1450 1455Arg Val Val Asp Thr Asp Ser Thr Gly Ala Gly Glu Leu Val Glu1460 1465 1470Met Leu Asp Pro Asn Gly Thr Val Leu Ile Thr Gly Gly Thr Gly1475 1480 1485Ala Leu Ala Ala Glu Thr Ala Arg His Leu Val Glu Arg His Lys1490 1495 1500Ala Gly Arg Leu Leu Leu Val Ser Arg Arg Gly Ala Glu Ala Pro1505 1510 1515Gly Ala Ala Glu Leu Val Ala Glu Leu Ala Ala Leu Gly Ala Glu1520 1525 1530Val Thr Val Arg Ala Cys Asp Val Ala Asp Arg Asp Ala Leu Arg1535 1540 1545Arg Leu Leu Gly Glu Leu Pro Ala Glu His Pro Leu Ser Cys Val1550 1555 1560Val His Thr Ala Gly Val Leu Asp Asp Gly Val Leu Ser Ala Gln1565 1570 1575
Thr Thr Glu Arg Ile Asp Ala Val Leu Arg Pro Lys Val Asp Ala1580 1585 1590Ala Val His Leu Asp Gln Leu Thr Arg Glu Leu Gly Pro Val Pro1595 1600 1605Leu Val Leu Tyr Ser Ser Val Ser Ala Ser Leu Gly Ser Ala Gly1610 1615 1620Gln Ala Gly Tyr Ala Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala1625 1630 1635Ala Arg Arg Arg Ala Asp Gly His Pro Ala Leu Ser Leu Gly Trp1640 1645 1650Gly Trp Trp Ala Gly Ala Gly Met Ala Thr Gly Leu Glu Gly Ala1655 1660 1665Asp Ala Ala Arg Ile Arg Arg Ser Gly Ile Val Pro Leu Asp Pro1670 1675 1680Ala Asp Ala Leu Glu Leu Leu Asp Arg Ala Leu Ala Arg Pro Glu1685 1690 1695Pro Ala Leu Leu Pro Val Arg Leu Asp Leu Pro Ala Leu Arg Ala1700 1705 1710Ala Ala Arg Ala Thr Ala Pro Pro Glu Val Leu Arg Glu Leu Ala1715 1720 1725Gly Val Pro Ala Asp Ser Gly Ala Ala Leu Gly Ala Gly Gly Arg1730 1735 1740Val Gly Asn Gly Gln Arg Pro Asp Pro Ala Ser Pro Ala Glu Ala1745 1750 1755Leu Ala Ala Arg Leu Ala Pro Arg Ser Ala Ala Glu Arg Thr Ala
1760 1765 1770Leu Leu Leu Asp Leu Val Arg Ala Glu Val Ala Ala Val Leu Gly1775 1780 1785His Gly Asp Pro Ala Ala Val Gly Ala Gly Arg Ser Phe Lys Asp1790 1795 1800Ala Gly Phe Asp Ser Leu Thr Ala Val Asp Leu Arg Asn Arg Leu1805 1810 1815Asn Ala Arg Thr Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp1820 1825 1830His Pro Thr Pro Leu Ser Leu Ala Glu Leu Leu Arg Ala Asp Leu1835 1840 1845Glu Ala Ala Gly Leu Val Gly Ala Thr Gly Pro Ala Thr Gly Glu1850 1855 1860Pro Thr Gly Pro Glu Asp Leu Ser Ser Val Leu Asp Arg Leu Glu1865 1870 1875Ser Ser Leu Thr Ala Thr Asp Asn Gly Asp Ala Arg Ser Ala Ala1880 1885 1890Ala Arg Arg Leu Cys Ser Leu Leu Ala Met Leu Thr Ala Gly Ser1895 1900 1905Gly Glu His Pro Gly Gln Gly Ser Gly Glu Ser Pro Arg Gly Ser1910 1915 1920Gly Asp Ala Val Leu Asp Arg Leu Gln Ser Ala Ser Asp Asp Asp1925 1930 1935Leu Phe Asp Leu Phe Asp Ser Asp Phe Gln1940 1945
<210>9<211>20<212>DNA<213>Artificial<220>
<223>Synthetic primer<400>9gtgatgtatg acgactacgg 20<210>10<211>20<212>DNA<213>Artificial<220>
<223>Synthetic primer<400>10aaacctcgga agtgtggtct 20<210>11<211>20<212>DNA<213>Artificial<220>
<223>Synthetic primer<400>11atcgagggcg tcggcggtac 20
權(quán)利要求
1.產(chǎn)麥迪霉素的放線菌，其含有麥迪霉素生物合成基因或其同源物，其中麥迪霉素生物合成基因的聚酮合成酶基因中的至少一個組件或至少一個組件的部分序列被取代以便編碼其它組件的相應(yīng)氨基酸序列。
2.根據(jù)權(quán)利要求1的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中的KS3β-酮脂酰?；d體蛋白合成酶的氨基酸序列的核苷酸序列被全部或部分取代以便編碼ORF1中的KS2β-酮脂酰?；d體蛋白合成酶的相應(yīng)氨基酸序列。
3.根據(jù)權(quán)利要求2的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中KS3的第157至第420位的氨基酸序列的核苷酸序列被部分或全部取代以便編碼ORF1中KS2的第2705至第2968位的部分或全部的相應(yīng)氨基酸序列。
4.根據(jù)權(quán)利要求3的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中KS3的第157至第254位的氨基酸序列的核苷酸序列被部分或全部取代以便編碼ORF1中KS2的第2705至第2802位的部分或全部的相應(yīng)氨基酸序列。
5.根據(jù)權(quán)利要求4的產(chǎn)麥迪霉素的放線菌，其中編碼麥迪霉素合成酶基因的ORF2中KS3的第157至第186位的氨基酸序列的核苷酸序列被部分或全部取代以便編碼ORF1中KS2的第2705至第2734位的部分或全部的相應(yīng)氨基酸序列。
6.根據(jù)權(quán)利要求1到5任一項的產(chǎn)麥迪霉素的放線菌，其中的放線菌為Streptomyces mycarofaciens。
7.根據(jù)權(quán)利要求6的產(chǎn)麥迪霉素的放線菌，其中的放線菌為保藏號為FERM BP-10501的Streptomyces mycarofaciens 1149-38菌株。
8.產(chǎn)麥迪霉素的放線菌，其是通過對權(quán)利要求1到5任一項所述的產(chǎn)麥迪霉素的放線菌進一步突變獲得的。
9.產(chǎn)生麥迪霉素的方法，其包括培養(yǎng)權(quán)利要求1到5任一項所述的產(chǎn)麥迪霉素的放線菌；以及分離產(chǎn)生的麥迪霉素。
10.產(chǎn)生基本上不含白霉素的麥迪霉素的方法，其包括培養(yǎng)權(quán)利要求6或7所述的產(chǎn)麥迪霉素的放線菌；以及分離產(chǎn)生的麥迪霉素。
全文摘要
本發(fā)明涉及提高了麥迪霉素產(chǎn)率的麥迪霉素高產(chǎn)菌株以及利用該菌株生產(chǎn)麥迪霉素的方法，所述麥迪霉素是大環(huán)內(nèi)酯抗生素的一個成員。產(chǎn)麥迪霉素的放線菌含有麥迪霉素生物合成基因或其同源物，其中麥迪霉素生物合成基因的聚酮合成酶基因中的至少一個組件或至少一個組件的部分序列被取代以便編碼其它組件的相應(yīng)氨基酸序列。
文檔編號C12R1/04GK1896226SQ200610073849
公開日2007年1月17日 申請日期2006年3月31日 優(yōu)先權(quán)日2005年3月31日
發(fā)明者渡邊學(xué), 中橋正晃 申請人:明治制菓株式會社