CN101275141A - 阿嗪霉素的生物合成基因簇 - Google Patents
阿嗪霉素的生物合成基因簇 Download PDFInfo
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Abstract
本发明是一种由链霉菌产生的具有抗肿瘤活性的抗生素——阿嗪霉素(Azinomycin B)的生物合成基因簇的克隆、测序、分析、功能研究及其应用。整个基因簇共包含34个基因:1个重复使用的I型聚酮合成酶基因;2个萘环修饰酶基因;8个非核糖体聚肽骨架合成及修饰酶基因;11个非天然氨基酸结构单元合成酶基因;1个抗性基因;3个后修饰酶基因以及8个功能不确定的基因。通过对上述生物合成基因的遗传操作可阻断阿嗪霉素的合成;对萘环合成和修饰基因的异源表达可产生其前体化合物。本发明所提供的基因及其蛋白也可以用来寻找和发现可用于医药、工业或农业的化合物或基因、蛋白。
Description
技术领域:
本发明属于微生物基因资源和基因工程领域,具体涉及抗肿瘤抗生素阿嗪霉素(Azinomycin B)的生物合成基因簇的克隆、分析、功能研究及其应用。
技术背景:
阿嗪霉素(Azinomycins)是一种能够诱导DNA链间交联的天然抗生物,由链霉菌Streptomyces sahachiroi NRRL2485所产生,结构十分独特。它含有两个活性很强的反应中心:氮杂双环(1-azabicyclo[3.1.0]hexane)的脱水氨基酸残基和环氧三元环。其中的氮环结构仅在天然化合物Ficellomycin中发现[J.Antibiot.(1989)42,357-360]。作为一类小分子化合物,阿嗪霉素可以结合在DNA的大沟,选择性地作用于序列5’-d(PuNpy)-3’[J.Am.Chem.Soc.(2002)124,13008-13017],通过氮杂环上的C-10原子对DNA嘌呤氮的亲电子攻击形成烷基化的单聚络合物,进而启动环氧杂环***的C-21原子对DNA互补链上鸟嘌呤G的N-7原子进行亲电子攻击,最终使DNA双链发生链间交联。阿嗪霉素具有良好的抗肿瘤活性,组分A和组分B在体外对于肿瘤细胞株L5178Y的IC50抑制率分别为0.07和0.11ug/ml[J.Antibiot.(1986)39,1527-1532]。在体内一系列实体瘤的生长抑制方面,B组分要优于A组分,其治疗效果与临床上使用的ISC抗生物丝裂霉素C相当,但剂量更低(如能使肿瘤小鼠45天的存活率达到57%和生命期延长200%左右的阿嗪霉素B和丝裂霉素C的用量分别为32ug/kg/day和1mg/kg/day)[J.Antibiot.(1987)40,60-65]。由于阿嗪菌素所诱导细胞DNA的ISC反应和烷基化反应受pH影响很大,在pH较低的反应环境中发生较快;而肿瘤细胞相对于正常细胞,通常由于产生过多的乳酸导致pH较低,这就使阿嗪霉素在肿瘤治疗过程中具有一定的选择性[J.Biochem.(1977)55,630-635]。但是,阿嗪霉素化学结构的不稳定严重限制了它们在临床上的应用[J.Antibiot.(1986)39,1527-1532]。
鉴于阿嗪霉素良好的生物活性,近40种通过化学合成获得的结构类似物被用于大量肿瘤细胞株的活性测试,包括部分药物抗性细胞株[Tetrahedron(2001)57,4467-4488;Bioorg.Med.Chem.Lett.(2005)15,653-656]。研究发现,只保留环氧***的结构类似物仍然具有很高的细胞毒性(只发生DNA的烷基化,不引起ISC效应),而只有氮杂双环的结构类似物活性急剧下降,表明氮杂双环的化学结构虽然对于DNA的烷基化十分重要,但并非阿嗪霉素生物活性所必须[Bioorg.Med.Chem.Lett.(2000)10,239-241;Biochemistry(2000)39,14968-14975;Angew.Chem.Int.Ed.(2000)39,3467-3470]。同时,阿嗪霉素的抗肿瘤活性与环氧***两侧的萘甲酸和氨基边链取代基的微小变化都会引起活性的明显变化,显示两者对活性中心环氧***发生的效应有增强作用[Org.Lett.(2002)4,3545-3548]。
构效关系研究显示,诱导ISC的发生并非阿嗪霉素及其类似物产生细胞毒性的唯一途径,可能还存在由环氧***所介导的其它细胞效应方式。一系列具有较好生物活性的类似物的获得,表明阿嗪霉素是一类很好的可用于新药开发的先导化合物。阿嗪霉素的药效基团和其它化学结构的重新组合,可能创造出更有临床应用价值的新型药物。但是,由于过于集中的化学官能团和结构的不稳定,阿嗪菌素的获得是合成化学的巨大挑战。自1986年首次发现以来,2001年才首次完成了阿嗪霉素A的化学合成[Tetrahedron(2001)57,4467-4488;J.Am.Chem.Soc.(2001)40,1736-1739]。通过化学合成所获得的结构类似物即使在活性和结构稳定性方面的性质得到改善,也可能因为合成的复杂性而使实际生产成本过高。
我们以微生物来源的阿嗪霉素为目标分子,从克隆生物合成基因簇出发,采用微生物学、分子生物学、生物化学及有机化学相结合的方法研究其生物合成,通过对其生物合成机制的研究揭示包括氮杂双环在内的独特化学结构形成的酶学机理,在此基础上运用代谢工程的原理,合理修饰阿嗪霉素的生物合成途径,探索结构稳定、活性更好、并能通过微生物发酵大量生产的新型药物。
发明内容:
本发明涉及一种由链霉菌Streptomyces sahachiroi NRRL2485产生的具有抗肿瘤活性的抗生素-阿嗪霉素(Azinomycin)的生物合成基因簇的克隆、测序、分析、功能研究及其应用。
本发明中整个基因簇共包含34个基因的核苷酸序列或互补序列(序列1),其中1个(aziB)用于编码重复使用的I型聚酮合成酶(PKS),共包含5个功能域,负责萘甲酸基团的生物合成;2个基因(aziB1和aziB2)编码的氧化酶和甲基转移酶负责对萘环进行修饰;8个基因(aziA1,aziA2,aziA3,aziA4,aziA5,aziA6,aziA7,aziA8)负责合成非核糖体聚肽骨架合成及修饰;11个基因(aziC1,aziC2,aziC3,aziC4,aziC5,aziC6,aziC7,aziC8,aziC9,aziC10,aziC11)负责非天然氨基酸结构单元的合成酶;还包括1个抗性基因(aziE)、3个后修饰酶基因(aziD1,aziD2,aziD3)以及8个功能不确定的基因(aziH1,aziH2,aziH3,aziF,aziG,aziU1,aziU2,aziU3)。
本发明还提供了一个编码包含AL,KR,PCP非核糖体聚肽合成酶结构域的的苷酸序列,由序列2中的氨基酸序列组成,命名为aziA3,其基因的核苷酸序列位于序列1中第2596-6387碱基处。
本发明还提供了一个硫酯酶的核苷酸序列,由序列3中的氨基酸序列组成,命名为aziA6,其基因的核苷酸序列位于序列1中第6384-7205碱基处。
本发明还提供了一个编码包含C,A,PCP非核糖体肽合成酶结构域的核苷酸序列,由序列4中的氨基酸序列组成,命名为aziA4,其基因的核苷酸序列位于序列1中第7202-10513碱基处。
本发明还提供了一个编码硫酯酶的核苷酸序列,由序列5中的氨基酸序列组成,命名为aziA7,其基因的核苷酸序列位于序列1中第10796-11548碱基处。
本发明还提供了一个编码O-甲基转移酶的核苷酸序列,由序列6中的氨基酸序列组成,命名为aziB2,其基因的核苷酸序列位于序列1中第11548-12585碱基处。
本发明还提供了一个编码细胞色素P450氧化酶的核苷酸序列,由序列7中的氨基酸序列组成,命名为aziB1,其基因的核苷酸序列位于序列1中第12587-13792碱基处。
本发明还提供了一个编码包含AL,PCP非核糖体肽合成酶结构域的核苷酸序列,由序列8中的氨基酸序列组成,命名为aziA1,其基因的核苷酸序列位于序列1中第13873-15756碱基处。
本发明还提供了一个编码转运蛋白的核苷酸序列,由序列9中的氨基酸序列组成,命名为aziE,其基因的核苷酸序列位于序列1中第15839-17239碱基处。
本发明还提供了一个编码包含C,PCP,C非核糖体肽合成酶结构域的核苷酸序列,由序列10中的氨基酸序列组成,命名为aziA2,其基因的核苷酸序列位于序列1中第17541-20888碱基处。
本发明还提供了一个编码硫酸腺苷酰转移酶亚单位1的核苷酸序列,由序列11中的氨基酸序列组成,命名为aziH2,其基因的核苷酸序列位于序列1中第20878-22185碱基处。
本发明还提供了一个编码硫酸腺苷酰转移酶亚单位2的核苷酸序列,由序列12中的氨基酸序列组成,命名为aziH1,其基因的核苷酸序列位于序列1中第22185-22997碱基处。
本发明还提供了一个编码腺苷酰硫酸激酶的核苷酸序列,由序列13中的氨基酸序列组成,命名为aziH3,其基因的核苷酸序列位于序列1中第23101-23658碱基处。
本发明还提供了一个编码硫酯酶的核苷酸序列,由序列14中的氨基酸序列组成,命名为aziG,其基因的核苷酸序列位于序列1中第23726-24127碱基处。
本发明还提供了一个编码MbtH类似蛋白的核苷酸序列,由序列15中的氨基酸序列组成,命名为aziF,其基因的核苷酸序列位于序列1中第24328-24516碱基处。
本发明还提供了一个编码O-酰基转移酶的核苷酸序列,由序列16中的氨基酸序列组成,命名为aziD1,其基因的核苷酸序列位于序列1中第24547-25758碱基处。
本发明还提供了一个编码PCP/ACP的核苷酸序列,由序列17中的氨基酸序列组成,命名为aziC11,其基因的核苷酸序列位于序列1中第25755-26018碱基处。
本发明还提供了一个编码酮基转移酶C-端亚单位的核苷酸序列,由序列18中的氨基酸序列组成,命名为aziC6,其基因的核苷酸序列位于序列1中第26021-27043碱基处。
本发明还提供了一个编码酮基转移酶N-端亚单位的核苷酸序列,由序列19中的氨基酸序列组成,命名为aziC5,其基因的核苷酸序列位于序列1中第27040-27975碱基处。
本发明还提供了一个编码氨基转移酶的核苷酸序列,由序列20中的氨基酸序列组成,命名为aziC7,其基因的核苷酸序列位于序列1中第27972-29294碱基处。
本发明还提供了一个编码N-己酰-γ-谷氨酰磷酸还原酶的核苷酸序列,由序列21中的氨基酸序列组成,命名为aziC4,其基因的核苷酸序列位于序列1中第29735-30658碱基处。
本发明还提供了一个编码N-己酰谷氨酸激酶的核苷酸序列,由序列22中的氨基酸序列组成,命名为aziC3,其基因的核苷酸序列位于序列1中第30737-31615碱基处。
本发明还提供了一个编码N-己酰-鸟氨酸/N-己酰-赖氨酸脱酰基酶的核苷酸序列,由序列23中的氨基酸序列组成,命名为aziC10,其基因的核苷酸序列位于序列1中第31608-32738碱基处。
本发明还提供了一个编码己酰-CoA脱氢酶/还原酶的核苷酸序列,由序列24中的氨基酸序列组成,命名为aziC8,其基因的核苷酸序列位于序列1中第32741-33559碱基处。
本发明还提供了一个编码包含C,A,PCP,RE非核糖体肽合成酶结构域的核苷酸序列,由序列25中的氨基酸序列组成,命名为aziA5,其基因的核苷酸序列位于序列1中第33638-38261碱基处。
本发明还提供了一个编码包含KS,AT,DH,KR,ACP结构域的重复使用的I型聚酮合成酶的核苷酸序列,由序列26中的氨基酸序列组成,命名为aziB,其基因的核苷酸序列位于序列1中第38515-43854碱基处。
本发明还提供了一个编码未知蛋白的核苷酸序列,由序列27中的氨基酸序列组成,命名为aziU1,其基因的核苷酸序列位于序列1中第44054-44827碱基处。
本发明还提供了一个编码未知蛋白的核苷酸序列,由序列28中的氨基酸序列组成,命名为aziU2,其基因的核苷酸序列位于序列1中第44838-45503碱基处。
本发明还提供了一个编码未知蛋白的核苷酸序列,由序列29中的氨基酸序列组成,命名为aziU3,其基因的核苷酸序列位于序列1中第45500-46513碱基处。
本发明还提供了一个编码赖氨酸生物合成酶的核苷酸序列,由序列30中的氨基酸序列组成,命名为aziC2,其基因的核苷酸序列位于序列1中第46593-47456碱基处。
本发明还提供了一个编码分枝氨基酸氨基转移酶的核苷酸序列,由序列31中的氨基酸序列组成,命名为aziC1,其基因的核苷酸序列位于序列1中第47453-48184碱基处。
本发明还提供了一个编码细胞色素P450氧化酶的核苷酸序列,由序列32中的氨基酸序列组成,命名为aziC9,其基因的核苷酸序列位于序列1中第48254-49513碱基处。
本发明还提供了一个编码硫酯酶的核苷酸序列,由序列33中的氨基酸序列组成,命名为aziA8,其基因的核苷酸序列位于序列1中第49525-50274碱基处。
本发明还提供了一个编码己酰-CoA脱氢酶的核苷酸序列,由序列34中的氨基酸序列组成,命名为aziD3,其基因的核苷酸序列位于序列1中第50308-51456碱基处。
本发明还提供了一个编码分枝氨基酸氨基转移酶的核苷酸序列,由序列35中的氨基酸序列组成,命名为aziC2,其基因的核苷酸序列位于序列1中第51462-52463碱基处。
序列1的互补序列可根据DNA碱基互补原则随时得到。序列1的核苷酸序列或部分核苷酸序列可以通过聚合酶链式反应(PCR)或用合适的限制性内切酶酶切相应的DNA或使用其他合适的技术得到。本发明提供了得到至少包含部分序列1中DNA序列的重组DNA质粒的途径。
本发明还提供了产生阿嗪霉素生物合成基因被中断或加倍的微生物体的途径,至少其中之一的基因包含有序列1中的核苷酸序列。
本发明所提供的核苷酸序列或部分核苷酸序列,可利用聚合酶链式反应(PCR)的方法或包含本发明序列的DNA作为探针以Southern杂交等方法从其他生物体中得到与阿嗪霉素生物合成基因相似的基因。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的克隆DNA可用于从链霉菌Streptomyces sahachiroi NRRL2485基因组文库中定位更多的文库质粒。这些文库质粒至少包含本发明中的部分序列,也包含有Streptomyces sahachiroiNRRL2485基因组中以前邻近区域未克隆的DNA。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列可以被修饰或突变。这些途径包括***、置换或缺失,聚合酶链式反应,错误介导聚合酶链式反应,位点特异性突变,不同序列的重新连接,序列的不同部分或与其他来源的同源序列进行定向进化(DNA shuffling),或通过紫外线或化学试剂诱变等。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的克隆基因可以通过合适的表达体系在外源宿主中表达以得到相应的酶或其他更高的生物活性或产量。这些外源宿主包括链霉菌、假单孢菌、大肠杆菌、芽孢杆菌、酵母、植物和动物等。
本发明所提供的氨基酸序列可以用来分离所需要的蛋白并可用于抗体的制备。
包含本发明所提供的氨基酸序列或至少部分序列的多肽可能在去除或替代某些氨基酸之后仍有生物活性甚至有新的生物学活性,或者提高了产量或优化了蛋白动力学特征或其他致力于得到的性质。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的基因或基因簇可以在异源宿主中表达并通过DNA芯片技术了解它们在宿主代谢链中的功能。
包含本发明所提供的核苷酸序列编码的蛋白可以催化合成3-氧甲基-5-甲基萘甲酸及阿嗪霉素非天然氨基酸聚肽骨架,进一步催化合成抗生素。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的基因或基因簇可以通过遗传重组来构建重组质粒以获得新型生物合成途径,也可以通过***、置换、缺失或失活进而获得新型生物合成途径。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的克隆基因或DNA片段可以通过中断阿嗪霉素生物合成的一个或几个步骤而得到新的阿嗪霉素结构类似物或前体。包含DNA片段或基因可以用来提高阿嗪霉素或其衍生物的产量,本发明提供了在基因工程微生物中提高产量的途径。
包含本发明所提供的非核糖体聚肽合成酶可以通过缺失、***或失活来自于相同或不同的非核糖体聚肽合成酶***的一个或多个非核糖体聚肽合成酶结构域、模块或基因而产生新的聚肽化合物。
包含本发明所提供的聚酮合成酶可以通过缺失、***或失活来自于相同或不同的聚酮合成酶***的一个或多个聚酮合成酶结构域、模块或基因而产生新的聚酮化合物。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的片段或基因可以用来构建非核糖体聚肽合成酶库或非核糖体聚肽合成酶衍生库或组合库。
包含本发明所提供的核苷酸序列或至少部分核苷酸序列的片段或基因可以用来构建聚酮合成酶库或聚酮合成酶衍生库或组合库。
本发明所提供的催化合成5-甲基萘甲酸和3-氧甲基-5-甲基-萘甲酸的基因可用于合成5-甲基萘甲酸和3-氧甲基-5-甲基-萘甲酸
本发明所提供的阿嗪霉素骨架的后修饰基因提供了通过遗传修饰得到类似物的途径,所包含的氧化还原反应也可有其他应用。
总之,本发明所提供的包含阿嗪霉素生物合成相关的所有基因和蛋白信息可以帮助人们理解阿嗪霉素类天然产物的生物合成机制,为进一步遗传改造提供了材料和知识。本发明所提供的基因及其蛋白质也可以用来寻找和发现可用于医药、工业或农业的化合物或基因、蛋白。
附图说明:
图1:阿嗪霉素的化学结构
图2:阿嗪霉素生物合成基因簇的基因结构和限制性内切酶谱。(A)5个交叠的粘粒代表了抗生链霉菌基因组80kb的DNA区域,B代表限制性内切酶BamHI,实体表示已被DNA测序的部分,Probe-P1和P2代表标记的探针部分;(B)阿嗪霉素生物合成基因簇的基因组成。
图3:提出的阿嗪霉素各组成单元的生物合成途径。(A)3-甲氧基-5-甲基萘甲酸;(B)α-酮异戊酸;(C)氮丙啶[1,2a]吡啶氨基酸;(D)NRPS骨架合成和后修饰。
图4:基因置换与基因互补突变菌株发酵产物的生物活性检测和高效液相色谱(HPCL)分析
(A)生物活性检测和(B)HPLC分析;(1)野生型发酵产物;(2)PKS基因置换的突变体(ΔaziB)发酵产物;(3)NRPS基因置换的突变体(ΔaziA3)发酵产物;(4)aziC7基因置换的突变体(ΔaziC7)发酵产物;
图5:在S.albus中异源表达阿嗪霉素的生物合成基因
(A)含有空载体pTGV2的S.albus发酵产物;(B)含有aziB的突变株的发酵产物;(C)3-甲氧基-5-甲基-萘甲酸标准品;(D)含有aziB,aziB1和aziB2的突变株发酵产物。
图6:细菌中的重复使用的I型PKS
(A)结构域组成和机制(I)AviM和CalO(II)ChlB1和MadB(III)NcsB(IV)AziB;(B)AviM,CalO5,ChlB1,,MadB,,NcsB和AziB的同源性比较。
符号说明:
图1
Azinomycin:阿嗪霉素。
图2
Probe:探针Skeleton Assembly:骨架生成;Naphthoate:萘甲酸;Tailoring:后修饰;Building Blocks from amino acid;Functional Unassigned:功能不确定;Resistance:抗性;Unknown Function:未知功能;Beyond the Cluster:基因簇之外。
图3
AL:酰基辅酶A连接酶;PCP:肽酰载体蛋白;C:缩合酶;KR:酮基还原酶A:腺苷化酶;RE:还原酶。
图6
KS:酮基合成酶;AT:酰基转移酶;DH:脱氢酶;KR:酮基还原酶;ACP:酰基载体蛋白;2-hydroxyl-5-methyl-NPA:2-羟基-5-甲基-萘甲酸;5-methyl-NPA:5-甲基-萘甲酸。
具体实施方式:
以下结合图1-图6对本发明进一步详细说明。
1.克隆阿嗪霉素的生物合成基因片断:
尽管在阿嗪霉素的化学合成和作用机制方面的研究很多,但有关其生物合成起源方面的认识却非常少。近年来,采用13C-标记的乙酸对阿嗪霉素的生物合成进行同位素标记喂养实验表明(Chem.Commun.(2004)8,990-991;Chem.Commun.(2004)22,2600-2601),其萘甲酸结构单元可能来源于聚酮的生物合成途径,以己酰-CoA为起始,在聚酮合成酶(PKS)的催化下,于5个丙二酰-CoA之间的连续缩合反应形成线形的聚酮中间产物,经过醛缩反应成环,脱水、发生甲基取代最终形成萘甲酸的结构单元。标记实验同时显示,烯醇残基可能来源于苏氨酸,含有氮杂双环的脱水氨基酸残基则可能来源于α-酮戊二酸衍生物(如谷氨酸)。此外,推测环氧残基可能来源于缬氨酸。由于氨基酸残基的高度修饰,包含环氧***和氮杂双环的短肽结构单元可能是由非核糖体聚肽合成酶(NRPS)催化形成。
阿嗪霉素的萘甲酸结构单元与新制癌菌素(neocarzinostatin)的萘甲酸结构单元非常相似。已有研究表明(Chem Biol(2005)12,293-302),新制癌菌素的萘甲酸结构单元是由重复使用的I型PKS所合成。根据仅有的几例重复使用的I型PKS的保守区域,设计了简并性引物5’-GCG GAC GGC TAC GGS MGNGGNGAR GG-3’和5’-CGA GCC GTG GCC SGA RAA NAC CCA NAC-3’,采用快速PCR的方法(Biochem Biophys Res Commun(2006)345:133-139),从阿嗪霉素产生菌的总DNA中克隆编码PKS基因部分序列,得到约0.8kb的PCR产物,克隆入pGEM-T Easy载体,经限制性内切酶分组,DNA顺序分析表明与已知的重复使用的I型PKS基因有很高的同源性。
2.阿嗪霉素生物合成基因簇的克隆,序列分析及功能分析:
将上述PKS基因片段用地高辛标记为探针Probe-1从Streptomyces sahachiroiNRRL2485的基因组文库约6000个克隆中筛选,分离得到的粘粒涵盖了染色体约50kb的区域。在此基础上经染色体步移得到了5个交叠的粘粒pAL1022,pAL1023,pAL10024,pAL1025,和pAL1026涵盖了染色体约80kb的区域(图2A)。DNA顺序分析了63,549bp的染色体区域,GC含量71.48%。生物信息学分析包含了47个开放读码框(图2B)。orf(-1)和详细的分析结果列于表1。
表1阿嗪霉素生物合成基因簇中各基因及编码蛋白的功能分析
3.阿嗪霉素生物合成基因簇边界的确定:
根据基因编码蛋白的功能分析,阿嗪霉素的生物合成基因簇被确定为从基因aziA3到aziD2(图2B),包含34个开放读码框。其中1个(aziB)用于编码重复使用的I型聚酮合成酶(PKS),共包含5个功能域,负责萘甲酸基团的生物合成;2个基因(aziB1和aziB2)编码的氧化酶和甲基转移酶负责对萘环进行修饰;8个基因(aziA1,aziA2,aziA3,aziA4,aziA5,aziA6,aziA7,aziA8)负责合成非核糖体聚肽骨架合成及修饰;11个基因(aziC1,aziC2,aziC3,aziC4,aziC5,aziC6,aziC7,aziC8,aziC9,aziC10,aziC11)负责非天然氨基酸结构单元的合成酶;还包括1个抗性基因(aziE)、3个后修饰酶基因(aziD1,aziD2,aziD3)以及8个功能不确定的基因(aziH1,aziH2,aziH3,aziF,aziG,aziU1,aziU2,aziU3)。基因中断及基因置换aziB,aziC7,和aziA3完全抑制了阿嗪霉素的产生(图4),进一步证明了克隆到基因簇为阿嗪霉素的生物合成基因簇。
4.萘甲酸结构单元的生物合成途径:
萘甲酸结构单元的生物合成如图3A所示。aziB编码的重复使用的聚酮合成酶(包括KS,AT,DH,KR,ACP结构域)催化1分子乙酰辅酶A和5分子丙二酰辅酶A缩合,脱除5分子水生成5-甲基萘甲酸。aziB1编码的P450羟化酶催化5-甲基萘甲酸在3-位的羟化反应,生成的羟基接着在aziB2编码的O-甲基转移酶催化下进行甲基化修饰,从而完成阿嗪霉素中3-甲氧基-5-甲基萘甲酸单元的生物合成。
5.来源于氨基酸的特殊结构单元的生物合成:
α-酮戊二酸推测由缬氨酸在aziC1所编码分枝氨基酸氨基转移酶的作用下发生脱氨基反应所产生(图3B),此过程与初级代谢中缬氨酸分解代谢的第一步反应一致。
阿嗪霉素中含有一个非常特别的氮杂双环[3.1.0]己烷的螺环体系,该结构的生物合成过程如图3C所示。aziC2编码的N-乙酰基转移酶催化谷氨酸α-氨基的乙酰化,接着在aziC3编码的N-乙酰谷氨酸激酶和aziC4编码的γ-谷氨酰磷酸还原酶的作用下生成N-乙酰谷氨酸γ-半醛。在aziC5与aziC6共同编码的转羟乙醛基酶的作用下,N-乙酰谷氨酸γ-半醛完成一次两碳单元的延伸生成2-乙酰氨基-5,7-二羟基-6-酮庚酸。aziC7编码的转氨酶对其进行转氨反应后,在aziC8编码的脱氢酶催化下形成α,β不饱和庚酸,此不饱和酸在6-氨基的亲核进攻下发生分子内Michael加成反应生成四氢吡咯环,随后的进一步的修饰作用形成阿嗪霉素中的氮杂双环结构,在此过程中还伴随着aziC9编码的P450羟化酶催化四氢吡咯环C4位的羟化反应。最后在aziC10编码的脱乙酰基酶的催化作用下脱去氨基上的乙酰基,从而生成阿嗪霉素中的氮杂双环[3.1.0]己烷的螺环结构单元。
6.阿嗪霉素骨架合成及后修饰反应
阿嗪霉素生物合成基因簇中有五个基因(aziA1-A5),它们负责编码阿嗪霉素骨架组装的相关酶(AziA1-A5)。AziA1包括两个功能域(AL-PCP),其中AL域负责将起始萘甲酸单元的活化并将其装载在PCP上。AziA3包含三个功能域(AL-KR-PCP),其中的AL与一些α-酮酸腺苷化域同源,它负责将α-酮异戊酸活化并将其装载在对应的PCP上,KR结构域对酮基进行还原,生成第一个延伸单元α-羟基异戊酰-PCP。AziA2包含三个功能域(C-PCP-C),它负责催化α-羟基异戊酰-PCP与起始单元的缩合。AziA4与AziA5则是两个典型的NRPS,它们分别含有三个功能域(C-A-PCP)和四个功能域(C-A-PCP-Re)。AziA5中的A与很多负责苏氨酸腺苷化的功能域同源,它负责苏氨酸的活化与装载,AziA4的A则负责特殊氮杂双环氨基酸的活化与装载。这两个氨基酸延伸单元在各自对应的C功能域催化下与上游的酰基发生缩合完成阿嗪霉素骨架的组装,最后在AziA5的Re功能域的作用下将组装好的酰基-PCP还原,以醛基的形式从NRPS上释放下来。释放的化合物进一步在AziD1,AziD2和AziD3的催化作用下分别发生乙酰化、氧化和环氧化反应最终形成完整的阿嗪霉素分子。整个过程如图3D所示。
7.阿嗪霉素生物合成基因簇的应用-生物合成基因簇中的基因可以在异源宿主S.albus中进行异源表达:
在克隆、分析了完整的阿嗪霉素生物合成基因簇,研究了各基因编码蛋白可能的功能的基础上,本发明对阿嗪霉素萘甲酸结构单元的生物合成机制进行了初步探讨。将推测的萘甲酸结构单元的合成基因-重复使用的I型PKS aziB基因置于ErmE*启动子下于S.albus中进行异源表达,得到的突变株经发酵、HPLC和LC-MS分析,显示aziB导入的突变株可产生5-甲基萘甲酸(图4II)。将我们推测的两个萘甲酸修饰酶AziB1和AziB2一起导入S.albus,产生了预期的3-甲氧基-5-甲基萘甲酸(图4IV),证实了我们对阿嗪霉素萘甲酸结构单元生物合成途径的推测。同时也证实阿嗪霉素生物合成基因簇中的基因可以在异源宿主S.albus中进行异源表达。
以下进一步提供实施实例,这些实施实例有助于理解本发明,仅用作说明而不限制本发明的应用范围。
实施例1
阿嗪霉素产生菌链霉菌Streptomyces sahachiroi NRRL2485总DNA的提取:
将100μL 1×108S.lavendulae 314孢子悬液接种到3mL TSB液体培养基中,30℃,230rpm培养约24hr后达到对数生长期后期,取2mL接种到50mL TSB(含10mM氯化镁,0.1%甘氨酸),30℃,250rpm培养约23hr后达到稳定生长期前期,呈乳黄色浑浊,将菌液4℃,3500rpm,离心15min收集菌丝,用裂解液洗涤,收集淡乳黄色菌丝0.5mL。向1mL菌丝中加入10mL裂解液(含溶菌酶5mg/mL)共四管,涡旋至均一,37℃水浴15mim。加入0.1mL蛋白酶K(10mg/mL,用裂解液新鲜配制),1mL 10%SDS,混匀后迅速放入70℃水浴15mim,呈澄清。置冰上冷却,加入2.5mL 5M KAc,冰上冷却15min。加入10mL饱和酚,混匀,10mL氯仿,混匀,12000rpm,4℃离心20min。用破口的枪头将水相吸出置于新的离心管,加等量的CHCl3-异戊醇(24∶1)抽提,12000rpm,4℃离心10min。用破口的枪头将水相吸出置于新的离心管,加2倍的无水乙醇,混匀,有大团的DNA出现。将其钩出置于新的离心管,加5mL70%乙醇洗涤,将液体倾出,用枪吸净,加5mL TE溶解,加RNase A使终浓度为50μg/mL,37℃温育0.5小时。依次用等体积的饱和酚抽提两次,CHCl3-异戊醇抽提两次,向水相中加入0.1体积的3M NaAc,2体积的无水乙醇,轻轻的混合充分,有絮状DNA出现。将四管DNA合并到两管(每管中有1mL 70%乙醇用于洗涤),将液体吸出,再加1mL无水乙醇洗涤,吸出乙醇,超净台中吹干,溶于适当体积的TE(pH 8.0)中。
实施例2
阿嗪霉素产生菌链霉菌Streptomyces sahachiroi NRRL2485遗传转移***的建立:
培养含有适当质粒的E.coli S17-1至OD6000.3-0.4,20mL LB培养液中的细菌细胞离心收集,用等体积的LB洗两次,重悬于2mL LB中,作为大肠杆菌供体细胞。取适量冻存于-80℃的Streptomyces sahachiroi NRRL2485的20%甘油孢子悬液500μL,用等体积的TES缓冲液(50mM TES Na,pH 8.0)洗两次,重悬于等体积的TES缓冲液,50℃热激10min使孢子萌发。再加等体积的TSB培养基,37℃温育2-5hr。离心重悬于0.5-1mL LB中作为链霉菌受体细胞。将不同浓度的受体细胞100μL与等体积的供体细胞混合直接涂布在含有10mMMgCl2的平板上,30℃温浴20hr后,采用无菌水轻轻洗涤平板表面以洗去大部分大肠杆菌,在每一平板的表面覆盖3mL含萘啶酮酸(终浓度为50ng/μL)和相应抗生素的LB软琼脂或1mL无菌水。30℃培养5天以上挑取接合子。
由于Streptomyces sahachiroi NRRL2485对硫链丝菌肽,阿泊拉霉素和红霉素都敏感,最后确定遗传转移所用抗生素的浓度:硫链丝菌肽25μg/mL,阿泊拉霉素50μg/mL,红霉素50μg/mL。IWL-4(可溶性淀粉1.0%,K2HPO40.1%,MgSO40.1%,NaCl 0.1%,(NH4)2SO40.2%,CaCO30.2%,FeSO40.0001%,MnCl2.6H2O 0.0001%,ZnSO40.0001%,酵母提取物0.05%,胰化蛋白胨0.1%,琼脂2.0%,pH 7.2)培养基最好,接合转移效率最高。在IWL-4培养基上,无论是在链霉菌中可以自主复制的质粒pKC1139,pHZ1358,还是位点特异性整合的质粒pSET152都可以得到相应的接合子。
实施例3
阿嗪霉素产生菌链霉菌Streptomyces sahachiroi NRRL2485基因组文库的构建:
首先通过一系列的稀释实验来确定Sau 3AI的用量,在此基础上大量酶切得到的DNA片段略大于40kb,脱磷。pOJ446先用Hpa I从两个cos序列中间切开并脱磷,然后再从多克隆位点处用BamHI切开,获得两个臂,与制备的40kb的DNA片段连接过夜。于冰上融化于-80℃取出地Promega Packagene exract,立即加入10ul连接产物,轻弹混匀,于室温(约22℃)放置3hr。加入445ul Phagebuffer,倒转混合;加入25ul氯仿终止反应,离心使氯仿沉于底部,4℃保存。将冻存于-80℃的菌株E.coli LE392涂布在LB培养基上复苏。调取单菌落接种于3ML LB培养基中(0.2%maltose10和mMMgSO4),37℃,220rpm振荡培养过夜,转接1%到50ML LB培养基中(0.2%maltose10和mMMgSO4),37℃,220rpm振荡培养至OD600=0.6-0.8。取5ul包装液,加入95ul phage buffer,稀释,再加入100ul E.Coli LE392(OD600=0.67),混匀,37℃,30min,涂于LB(含100ug/mlApramycin)平板上。37℃倒置培养过夜,测定噬菌斑形成单位(pfu)以估算文库的效价。取100ul包装液加入3.9ml phage buffer混匀,再加入4ml E.Coli LE392(OD600=0.72)菌液,室温,30min,加入4ml LB,37℃,75min;2500rpm,10min,去上清,剩余1-3ml涂于LB(含100ug/ml Apramycin)平板上。37℃倒置培养过夜。
平板长有超过20,000个克隆,用LB刮下,加入甘油(终浓度18%)和Apramycin(终浓度50ug/ml),分装,于-80℃保存。随机从平板中调取10个克隆,接种于LB培养基中培养,按大肠杆菌的质粒的碱法小量制备的方法抽提重组黏粒。用Bam HI鉴定,于0.8%的琼脂糖凝胶上电泳,根据限制酶切的电泳分析图谱,我们判断出黏粒被酶切产生的片断大小,加和这些片断,从而推算出整个黏粒的大小,实验表明每个黏粒的***片断约为40Kb左右。对于链霉菌而言,其染色体DNA的大小约为8Mb,如果***片断为20kb的文库效价是2000-5000cfu,就足以代表它的整个基因组。根据以上实验,我们建立的文库效价超过为10000pfu/μg DNA,***片断约为40kb左右,这表明我们建立的文库具有良好的质量,能够满足文库筛选的需要。
实施例4
阿嗪霉素产生菌链霉菌Streptomyces sahachiroi NRRL2485的发酵、产物分离纯化与鉴定:
取100ul Streptomyces sahachiroi NRRL2485孢子涂布在GYM(葡萄糖0.36%,酵母提取物0.4%,麦芽提取物1%,碳酸钙0.3%,琼脂2%,自来水,pH 6.8)平板上,30℃培养七天后,用接种一块(约1cm2)长满孢子的琼脂块接种到75%PS5+培养基(Pharmamedia 0.5%,可溶性淀粉0.5%,葡萄糖0.2%,Casein acid hydrolysate 0.5%,硫酸铵0.2%,赖氨酸0.05%,鸟氨酸0.05%,甘氨酸0.05%,pH7.0)中,24小时后,转接5ml到100mL(500mL锥形瓶)75%PS5+培养基。。转接15ml到100mL(500mL锥形瓶)25%PS5+培养基,30℃,250rpm培养72小时。离心收集上清,将二氯甲烷萃取两次,有机相用无水硫酸镁干燥,真空旋干后于-80℃保存。100ml发酵产物溶于100ul甲醇,取20μL经HPLC检测。
HPLC分析:
UV=218nm;柱子:Aglient Rp18 column,4.6×250mm;流动相条件:V=1mL/min;A=H2O,B=CH3CN
时间/min | 0 | 10 | 35 | 37 | 40 |
B/% | 20 | 20 | 80 | 20 | 20 |
实施例5
PCR克隆阿嗪霉素的生物合成基因:
PCR体系包含:DMSO(8%,v/v),MgCl2(25mM),dNTP(2.5mM),兼并性引物(40mM),Taq DNA聚合酶(2.5u)及适量模板Streptomyces sahachiroiNRRL2485总DNA。首先95℃,3min,1轮;然后94℃,1min,68℃,1min,72℃,2min,5轮;94℃,1min,65℃,1min,72℃,2min,30轮;最后72℃,10min,1轮。PCR结束后,1%琼脂糖电泳检查结果。低熔点胶回收预期大小的DNA片段,与pGEM T Easy vector连接,转化大肠杆菌DH5α感受态细胞,涂布在含有氨苄青霉素,IPTG(异丙基硫代-β-D-半乳糖苷)和X-gal(5-溴-4-氯-3-吲哚-β-D-半乳糖苷)的LB平板上进行蓝白斑筛选。挑取白色菌落过夜培养,抽提质粒,EcoRI酶切鉴定是否含有预期大小的DNA***片段。***有预期大小DNA片段的质粒测序。
实施例6
核酸分子杂交:
1)DIG DNA标记:将待标记的DNA用无菌水稀释至总体积15μL,沸水浴中加热变性10分钟,立即置于冰盐浴中冷却。接着加入2μL引物混合物,2μLdNTP混合物,1μL酶,混合均匀后,37℃水浴约16小时。加入0.8μL 0.8MEDTA(pH8.0)以终止反应,加入2.5μL 4M LiCl混合均匀,再加入75μL预冷的无水乙醇沉淀标记后的DNA,置于-80℃沉降40分钟。4℃,12000rpm离心20分钟收集DNA,用预冷的70%乙醇洗涤DNA沉淀,真空干燥后重新溶于50μLTE((pH 8.0)中。
2)DIG DNA探针标记后的质量检测:稀释标记的DNA探针,至以下六个梯度,1、10-1、10-2、10-3、10-4、10-5。稀释标记的对照DNA至浓度分别为以下浓度1μg/mL,100ng/mL,10ng/mL,1ng/mL,0.1ng/mL,0.01ng/mL。分别取1μL上述梯度的DNA样品点在杂交用的尼龙膜上,根据7)所述步骤进行显色反应,对比标记的DNA探针和DIG标记的对照DNA的显色强度以决定标记的DNA探针浓度。
3)菌落杂交(文库筛选)的膜转移:将保存于-80℃的基因文库稍融,取50μL,用450μLLB稀释得到10-1的稀释倍数,倍比稀释得到10-2,10-3,10-4,10-5,10-6。300μL涂平板(15cm×15cm,平板为LB/50μg/mL卡那霉素)。选取合适的比例,使每块平板约1200-1500个克隆。照选定的比例均匀涂布四块平板,37℃培养过夜。根据平板的大小剪取尼龙膜,小心地覆盖于平板表面不要产生气泡,做好位置标记,1分钟后取下尼龙膜置于干燥滤纸上,干燥10分钟直至菌落结合在尼龙膜上。原始的平板置于培养箱中4-5hr,使克隆重新生长作为原平板。将尼龙膜置于变性液(0.25M NaOH,1.5M NaCl)饱和的滤纸上15分钟(不要浸过膜),转移至中和液(1.0M Tris.HCl,1.5M NaCl,pH 7.5)饱和的滤纸上5分钟。转移至2×SSC(20×SSC储备液(L-1):NaCl,175.3g,柠檬酸钠,88.2g,pH=7.0)饱和的滤纸上自然风干。取下尼龙膜置于烘箱中,120℃固定45分钟。常温下于3×SSC/0.1%SDS溶液中振荡洗涤3小时,以除去细胞碎片。
4)Southern杂交的膜转移:DNA样品在适当浓度的琼脂糖凝胶上电泳至适当距离,做好标记。浸泡于400mL 0.25M HCl中脱嘌呤20分钟,使溴酚蓝变黄,用去离子水洗数次。室温下浸入碱性缓冲液(0.5M NaOH,1M NaCl)15分钟并轻轻振荡。换液一次继续浸泡凝胶20分钟,并轻轻振荡,去离子水洗三次。取一张每边都比凝胶大1mm的尼龙膜,用去离子水完全浸湿,做好标记。采用向上毛细管转移方法,用10×SSC转移缓冲液转移8-24hr。用2×SSC略微洗膜,120℃烘烤30分钟。
5)预杂交和杂交:预热杂交液(20mL/100cm2)至杂交温度68℃,放入杂交尼龙膜,轻轻振荡并保温30分钟。将DIG标记的DNA探针在沸水浴中变性5分钟,立即置于冰盐浴中冷却。冷却后,将DNA探针与合适体积的DIG杂交液
(2.5mL/100cm2)混合均匀。去除预杂交液并立即把DNA探针/DIG杂交液加入,轻轻振荡保持杂交温度64℃或68℃约16小时。
6)杂交后严紧洗脱:室温下用2×SSC/0.1%SDS漂洗两次,每次5分钟。68℃,用0.1×SSC/0.1%SDS振荡漂洗两次,每次15分钟。
7)显色反应和检测:严紧洗脱后的尼龙膜在洗涤缓冲液(0.1M马来酸,0.15MNaCl,pH=7.5,0.3%(v/v)Tween 20)中平衡1-5分钟,接着在封闭缓冲液(封闭试剂以10%的浓度溶于0.1M马来酸,0.15M NaCl,pH=7.5)中封闭30分钟,然后在抗体中浸泡30分钟。用洗涤缓冲液漂洗尼龙膜两次后,用检测缓冲液(0.1M Tris-HCl,0.1M NaCl,pH=9.5)中平衡2-5分钟,最后将尼龙膜置于10mL新配制的显色溶液[NBT(nitroblue tetrazolium chloride)溶于70%DMF,浓度为70mg/mL,BCIP(5-bromo-4-chloro-3-indolyl-phosphate)溶于水,浓度为50mg/mL。用时10mL显色溶液中加45μLNBT,35μL BCIP]中,置于黑暗中显色。显色合适后用去离子水漂洗以终止反应。
实施例7
基因中断突变菌株的获得:
将获得的转化子接种到TSB液体培养基(Am 25μg/ml)中,30℃振荡约28hr。取出200μl涂布在ISP-4(Am 50μg/ml)平板,30℃培养6-8天,收孢子,保存于-80℃;取出10μl在ISP-24(Am 50μg/ml)平板画线,37℃培养,放置2-3天。挑37℃整合生长的单菌落,接种至液体培养基ISP-4(Am 25μg/ml),37℃,振荡2-3天。取出涂布在ISP-4平板(Am 50μg/ml),37℃整合2-3天,收孢子,保存于-80℃。
基因中断或基因置换所用载体为pOJ260或pKC1139,基因置换用红霉素抗性基因替代目标基因中间的DNA片段。构建好的质粒通过实施例2所述属间接合转移的方式导入Streptomyces sahachiroi NRRL2485中得到双交换突变体,所得突变体通过Southern杂交在基因型上加以证明。
实施例8
基因在S.albus中表达及发酵产物分析:
将目标基因和红霉素启动子克隆到pTGV2载体。正确的质粒通过ET12567和S.Alus J1074的属间接合转移得到各突变体的异源表达菌株
将培养至对数生长期的细菌(约48hr)转接0.5%的菌液于R5A(蔗糖100g/l,硫酸钾0.25g/l,六水氯化镁10.12g/l,葡萄糖10g/l,Hy-case amino 0.1g/l,酵母提取物5g/l,Mops 21g/l,2ml R5 trace elements solution,调PH=6.85,高压灭菌)的液体培养基中继续培养120hr,把所有的发酵物(包括菌体和菌液)的PH调至2-3,超声15min(10s/50s);用滤纸滤去大部分菌体,用等体积的乙酸乙酯萃取两次,旋干,重旋于甲醇中。
HPLC分析:
UV=218nm;柱子:Aglient Rp18 column,4.6×250mm;流动相条件:V=1mL/min;A=H2O(1‰TFA),B=CH3CN(1‰TFA)
时间/min | 0 | 5 | 10 | 25 | 26 | 29 | 30 |
B/% | 10 | 10 | 20 | 70 | 95 | 95 | 10 |
以下根据本发明内容提供的基因和蛋白序列:
氨基酸/核苷酸序列表:
SEQUENCE LISTING
<110>中国科学院上海有机化学研究所
<120>阿嗪霉素(Azinomycin)生物合成基因簇
<130>说明书、权利要求书
<160>1
<170>PatentIn version 3.3
<210>1
<211>63549
<212>DNA
<213>链霉菌Streptomyces sahachiroi NRRL2485
<400>1
ggatcctggc cgacctggac atggagaccg gcaggctgac gtgggtcaac cgcggcgacc 60
atctgccgat cctgatccgt gacagccgct ggatcaccga cctgtcctgt cccccggccg 120
gtcccatggg agccgacctg ggcctgtccg tcaccgtggc caccgagcag ctggagcccg 180
gtgaccgact gctgctgtac acggacggca tcgtcgaagc ccgcgacaga cacggaacgg 240
agttcggacg ggaccgtttc gtcgacttca tccgccgcca tcactccggc cggcacaccc 300
tgcacgagac cctgcgccgg ctgatggccg ccgtcatgga acaccacgac ggcaagctcg 360
acgacgacgc cacggtcctg ctcaccgaat ggcgcggcgg ccaccagcgc gagctgaccc 420
cctgaagcgc cgcgcacgcg gaaggaaacg gagatgagca ccgatcagcc gcccgccccc 480
cgagaacaag agcgcgcccc gcacgtcgat gtgcaccgca ctcgtgccgg ccgccatctg 540
gtcacgttcc gaggggacct cgatctgcac accgctcctc tcctggccga caccctgcag 600
ccactgctgg ccaccgaggg tcacagcgtg ctcctggacc tgtccggcgt caccttcctg 660
gactccaccg gcctgacgtg cctgatcgcg gcctaccgca ccgccagaac caccggtgcc 720
cgcctggcgc tgatcgcgcc cggtgaacgc gtgcgccaca tgctggctct gaccggcgcg 780
gaccaggtcc tgcacagcta cccgaccccc gatgcggcac ccggctgacc gccccgcccc 840
ggcgatcaca tcagcgggcc ccgagggcct cgggccggga tgaggtctgc gggaggcgga 900
ggatcagctc cctggacacg ggcgccggcg gtcgcgggaa cggccgcgcg ttcggtcgcc 960
actgcgttcc tcgcggtgct cgtcgtgtcg tcgtcgccgt ggcgcgccga cggcagttgt 1020
gccgctcacc ttccggcggt ccgcacaact gccgtgtccc ggacggggtt ccgagcgcac 1080
tgtgccgcca ccacctcgag agccggcctg cggcctcacc tgcctccgcc gtccgcggac 1140
ctcattctga aacgctcagt tggcgggggc ctgagtcgct aggggtaacc ggctcgggac 1200
cggcgtcgag tgcggcgagc acggcgtcgc cgttcgccct cgccccctcg gtcagcgctc 1260
acccgatcac gcagccgtcc cggacagcaa gctcagcggc agttcggtcg tgtagcggca 1320
tcctgccgta cttcgagtgg gcgatgcgac gatcttccag agggtgatgc gacgatggtt 1380
ccgtgtacgg ggcaggcagt tcaggtcaac tcttcggcgt gttgccgggt ctgtgtcgac 1440
tgtgcagcga tcgatgtcgg ccagagacaa gtccgcaccg caatcatggg tggatccgag 1500
gagagccgga gagcccaggc gacagcgggg atcgcagctc tgccgtgggg tccgctcggg 1560
catgtcgcgc ggcgaaacgg ggcatccgtg gctcgatagc gggggtgtcc ggtcgaagac 1620
ccgcgatggt gtccgcagag tgtggggcag ggaagctgac agtgtgtgca gtctgtcgtg 1680
gtgtcagcaa acttcgggaa tgtcgatcac cgcaggtcgt ggctctgttg tggggagacg 1740
gttgcggtcc ggcaactcgt cgtatggggc acggacccgc gtgtctccga ggagagcgcg 1800
tatgctagtg gacccatgga ccctttgtga gccgtttact aaatcaacat agctttcgag 1860
cacaaagatg actacatttg gatcatgcga atcactctcg catgacccgg gggggggatg 1920
agtactcgtg tgtgacctcg ccaagggcgc tcgcgtcctc ggccgctacg tctcatacgg 1980
gtaacggccg catagaagac gatagcaggt atagtatgaa atggactatt acggtcgaaa 2040
cgtacggagg cgctgacgag gaaccggcga cgatcactgc agtgatgcag agcgttcccg 2100
gcggaccgcc acggatagtc gaactggtgg tcagagcctc ggagcagggc gcgatccccc 2160
ccagaggagc actcgacatc gacttcgaga tgctggctca ggcggtgagc ggggtcgctg 2220
ctgtgccctc ctccctctcc gagagcggat cggagagtca ccggcctgca agcgtgtccc 2280
cccctgcagc cgaagtacag aatcagcgcc cttaccgcaa gatgcccgac ccggacgagg 2340
tcagagaagt cttcctcaga ctcaggagtg tggggaaggt ggcgcagcac tacgacgtac 2400
ctcgttacac cgcacaggca tgggtggaca ggctgcgtcg tatgggacat ctcgaagaag 2460
cgaaatcgag caaccccaga ctgcgcggca aaggacgtcg gtaggaagtg tcaccggtgc 2520
gcgggacgac gcctggacct gtgcgctgaa cgtgccttcc gggccgtgcc acggcccgga 2580
aggcacgccg tttcctcagc cggcgatcct ggccgccagg gagcggaccg tcggggactc 2640
gtagagcgag cccaccgaga ccggactgtc cagctgcccg ctcagcgcct ccgcaagctg 2700
cacagccctg atcgaatcgc accccaggct gaagaacgag tcgtccctgc ccacgcgcgc 2760
gactccgagc acctcggccg cggcctcggc caacagtgct tcgagccgtc cttcgggcgg 2820
cagataggag gcgccgcccg cgccgacggc cgcgagtaca gcggccgggt cgggggtgcc 2880
actgcggtcc cgctctatcc gtgaggtcac cacgacccgg gcgaagatcc cacggtcggc 2940
cagttgagcc gacaccgcgg cccgcacctg ctgagggtcc gtgtccgacg cgggtacgac 3000
cgccacggcg acggtgccgc cgcccaattg gtccgccgcc aggagcggct tgatctgctc 3060
gttcgccgga tccgcgccga tgagcaggtg taccggcccg gcgtgcaggg cgcccagcag 3120
ggccagcgtg ccccgggtcg ggtcgatcag ggtcagtccc cggtgcctgc ccgccgcgac 3180
cagcggactg ccctggttca tccccgggcc gtcccacatg ctccaggcca ggctctgggc 3240
ccgtcgcccc gccgcggccc agcgctgggc ccagccgtcc agtaccgcgt tggccgccgc 3300
gtacgcaccg aaacccgtcc cgccgaacag accgttcacg gacgagtaca gcaccacggc 3360
ggtgtccggc cgtgtggcca gcaccccatc gagggccgca cagccaccgg ccttcggatg 3420
cagcatcgtc gcgagccact gcgcggactc ggcccgcagt tcgtggtcgg tcaaggactc 3480
ccactgcggg gcgaccgcgg caccggcgag atgcagcacc aggtcgaggc cgcaccccca 3540
ggacgcctcc gcttcggcca ccgccccggc cagcgcgtcc gggtccgcgg catcggccgt 3600
gcggtagcgg acctcgccga tggcgtgcag gacctccagc gcctcccggg ccccggtgtc 3660
caggcgctcc tgcggggtcc gcccgacgat cagcagccgt gccccgtgcg tgaccaccag 3720
gtgctccgcc acggctcgtc ccaggccgcc gagaccaccg gtgatgaggc aggtcccgcc 3780
ccggggcagc acatcagcgg gtacgtcgaa gacctccggc aacggcacgg cgcgcagccg 3840
ctgttgcagc ggcctgccgt cacgcacggc gatcacgtcg ccgctgtgcc gcaccgtggc 3900
cagggccgcg agctcggccg cgccggcgtc cgccggggcg tcgatcagtg cggtacgggc 3960
gagaagccgt tccgagcgcg cggtacgcac cagcgcggtc agtccggtcc ggcccggaac 4020
aaccgtgtcg cccggggcgg ccggcacggc tccccgggtc agcacggtga gctcggcatc 4080
ggggcggatc cgcgccacgg ccgacgccac ggcgaggaac tccgacagca ccccggcgtc 4140
gccgtcggcc ggcgccgtcg cggggtccga ggcggcgacg aacagcacct gtggtacggc 4200
cccttcgtcc gtgggcccct cgccggccgt cgcgaaggcc cgctccaggg ccgaggccag 4260
ctccacggcg tccgggccga ccacgatctg agccgtggtc gacacgggca gccagtcggg 4320
gacgtcaccg ctgacacagg ccacgagggg accgtccggc cgttgcctca gccgtgcgtc 4380
ggccggttcc cacacgaccg ccagcgcggg gacggaggcc tccgagtccg gcgcctcgtc 4440
cggtgcgacg ccgtccgtcc caccgccgta acagcggtcg tcgaacaggc cttcgcgcca 4500
gccttcgagc agccgctcgc gctgcacctt cccgccgttg tgccgtggga actcccggga 4560
cgtcaccggc accacgtagg tcggacgcag ggccaggtcg cgggcgagag ccgagcggat 4620
ggcgtccacc agcgcgggca cgtcgtcggt gccgggcacg aagaagacca ccagagcgtc 4680
ggtgccggtg tcctcgtcgg gcaccccggc caccgcggag caggcgggca cgacgccggg 4740
tacctgctcc accaccgcct cgatctcgtg cgcggggaag ttcgccccgt tgacgatcac 4800
catgttcttc cggcggccgg tgaggaacag ccggccgtca cgcaggaacc ccaggtcacc 4860
cgtgtcgaac cagccgtccg cggtgaaact ctcccggttg gcccgctcgt tgtacgcgta 4920
accgctcagg accgtgtcgc cggacacgtg cagccgcccg acccggccct ccgggagcac 4980
ctctccttcc tggtccacga tgcgcagggc gacaccgggc accggggaac cgacctcggt 5040
caccacgacc gcgccgggag tgcccgccgg cagctcgcgc agttcgccgt ccagcgaggc 5100
gggcaccagg gagacggtgc cgaccgccgc gtcgtcacgg tgcatgcggg tgtacgtgac 5160
ccccgaggac gtctccgaca tcccccagca gggcaccatg gcgtcggcgg gcagtgcgtg 5220
cggggcgagc agttccacga agcggagtgc ggtacgggcg acgacggcct caccggcgtt 5280
gcagatgttg gtcagacggc tgaggtccca ggaaccggcg gcgatctcct ccgcgcgctg 5340
gcacacgagc gagaacgcga agttgggcgc ccaggtgttg gtggcacgga accggtccag 5400
acagtccagc cacagcaacg gcctgcggat caccgactcc gttcgtacgt tgacgtgctc 5460
acacgccagg aacacgtccc ggaggttgta catgatcatg ccgccgacgt ggtccagcgg 5520
catccagttg agcgagacgt cgtgctccgt gaagccgttc gcggcgatcg ccgcgtagct 5580
cctggcgacg atcgtgcgat ggcggtgctg cacgcacttg ggcgtgccgg tgctgccgga 5640
ggtgagcagg ttgacgatca gcgcgtccgg gtccggtacg gtccagtcgg ccggttccgg 5700
atcgggcagg gcgtcggagg ccagtacccg cggcgcctcg ccgccaggca gtcggccggc 5760
cagcgaacgc accccttcgc acagcccggg atcggcgatc accggcggcc ggccgagcag 5820
gtcccaggcg gcggcgagct tgcgtaccgc ggcgttgtcc gtggcgtact ccggcgccgg 5880
agccagcgga gtgggcagga acccgcccag cacacaggcc caccaggcgg tgacgaacgt 5940
acggctgtcg gcgcactgca gcagcacggg gtcgcccggg gccagaccgt ggcggcgcag 6000
tccgcccagc atgcggcggg cgtcctcgta cagctcggcg tacgtctgcc ggtcggtgct 6060
gccgtccggc agcagatagg tggtgccgcg gcccggagcc acccgcgccg cccgggccag 6120
cgcctgcggc agcgtcggca cggcgtccgg cggcagcacc ggctcggggc cgcgaaccag 6180
ggccggacgg tcggcgggga cggcggtgcc gcgccccgct tcgcctccgc cggacccggg 6240
accggacgcc gcgcggcgcg tggcggtgac gcgcagccgg cggtcgcgga cggcggcggc 6300
gtcacggacc ccggcggtac cggccgcgat ctcggcgacg gtctccaggg agagctgggg 6360
gactgcggcg ctggtggcgg tggtcacgaa gaggtccttc cggccggagc gggcggttcg 6420
acggccagct gtccaccgat ctcccgggcg gcctccgggg tgagtccgac gacgaagtag 6480
tgcccgccct ccacactcag accggtccgg cccacggccg tctgcgcggc ccagccggcc 6540
aggtccgact cgtccaccga gccggtcagc ccccgaaccg tggtgatcgg aaccggcagc 6600
gggggacgcg gccggtgccg gtagttctcc cagagctcga agtcactgcg cagcagcggc 6660
aggagcatgt ccagcaacgg cgcgttgcgg gccaccgcgt cgggcagccc gccgcgctcg 6720
cgcaccgtcg ccagcagctc ctcgtccggg agcaggtgca acggctccgc aggcggcggc 6780
gcctgcggag cgggctgctc gccgacgatc agccgctgcg gccacgttcc gaaccggcgg 6840
cccatctcgt gcgcgatctc gtacgccagc aacccgcccg cacagtgccc gtacagcacg 6900
tacggcatgg tggtgtacgg gcgcagggcg atcgagcagg cggtgaccag cgtttcgatg 6960
cgggccggcg gctcctcccg ggagcggtcc tcacggccgg gcagctgcac cgtcagcgtc 7020
tccacccagg agggcatgag tgccgccgtc cgctggtaca cggacgcgcc gccaccggcg 7080
aacggcagac agatcagccg tacccgggcc tgttcgcggg ggacgcggcg cacccagggc 7140
gagtccatgg tgaagctcac cggtccgctg cgcggggacg cccctgcgtg ggtgtccgtg 7200
ctcaacgcgt cccctccccg tccagcgagg cggccagctc ggcgagttcc tcgtcgctgc 7260
tcccctccac caggtccagg gcctccgcca gcgcgtcccg ccgcgcttcc tggcccgaac 7320
gcgccgcgtc gacgagcccg gccagaccgg ccacggtggg agtggcgaag aagtcggcga 7380
tcggcagggc caccccgaac acggcgttca cgcgggagga cagctggatc gcctgaagac 7440
tggtgccgcc cgccgcgaag aagttcacgt gattgccgat ccgggcgccc agcagcgggg 7500
tgaacacctc ctcgcgcatc gtgcgctcgg tgggcgtgct gttctcctcg tcggcctcgg 7560
agagttcgtc caccacggcg aactccacgg cgggcagccg ccgtcggtcg atcttgccgc 7620
tcggggtcag cgggaccgcg tcgagcggca ccaggacgga ggggaccatc gcggccggca 7680
gccggcgcgc caccgcggag cggatcgtgt ccagctgcag ctcgccggga cgctccggca 7740
ccacgtacgc caccagcagc gcgccggtgc gcggatcggt cgccacctcc gccaccgcct 7800
gggcgactcc gtccaccgcc tgcagcgcgg cctccacctc gcccagctcg atccgcacgc 7860
cgcggacctt cacctgccgg tcggccctgc cgacgaagca caggtcccct gcctcgtccc 7920
agacggccag gtcaccggtg aggtagcggc gggaaccggg aggcccgtcg gggtccggcc 7980
ggaaccggtc ggcggtgagg gcgggctggc ccaggtagcc gcgcgccacg cccagcccgg 8040
cgatggcgag ttcaccgacc gcgccggccg ggagcaacga cgcaccggcg ccgtccacca 8100
cgtaggcacg gtgattggcc atcgcccggc cgatcggcgg cgaggccgtc cactcgccct 8160
cgcatcgctt ggccaccacg gtcaccgtgg tctccgtcgg gccgtacccg ttccagaagc 8220
tccggccacg ggcccaccgc gtggtcagct cgccggtgaa cgcctcgccc ccgacgaacg 8280
ccacccgcag gtcgggatag ttctcgggag cgagcagatc catgatcgcc gggggcaggt 8340
cgatgacggt gatccgctgc tcctgcagga ctgcgtcgag ggcgtccacg gagcgccgct 8400
ggtcctcgtc cagtacgtac aggaccgcgc cgctgagcag ggttccgaag atctcgaaca 8460
ccgagacgtc gaagccgggg gaggcgaaat gcaggacccg gtcctcggga gtgaggtcga 8520
acatgtcccg gacggtgcgg gtgaagttgg ccgcgttgcg gtgctcgacc accaccccct 8580
tgggacggcc ggtggacccc gaggtgtaga tgacgtacgc cgcggaggcg gccggcacga 8640
cgaccgcagg cggctcctcc tccgcctgcg gtccgtccag cggatcggca ccgacgacca 8700
gcgccagcgg accggggacc cggtccgccg taccggggga gaccaccacg gtcctggccc 8760
cggtgtcctc caggatcatc gccagccgct cgtcgggctg gccggcgtcc accggcacgt 8820
aggcgccccc ggcccgcagg acaccgagga cggcggcgag cagggccggg ccccgctgca 8880
tcaccaccgg cacgaggtcg ccgagcccca ccccggcggc cgtcagccgc cgcgcgatcc 8940
gccaggacca ggcgtccagt tcgccgtagg acagggagcc gcccttccac agcagcgcgg 9000
tgttgtccgg agccgccgcg acgacctcgg cgaacaggtc gtgcagcggg gtgtcggcag 9060
gcgcgccgat cggctcgttc cagcgctccc gtacgtcggc gtgctcctgc ggcgtgccca 9120
gcggggcgtc tcccagggga cgccgcggct gtcggcagag cgcgtccagg gtctgcagga 9180
aacggtccag gacgcgggtg gccgtggccc gccggtagag agaggtggcg aaccgcacgg 9240
ccacggtggg ctgttcaccg tccgtgacct ccagcgcgag ttcgaacgcg ccggacccgg 9300
cgtcgatctg ctcgcagccg aagcgcaccc cgtcggcctc ggccggtccg ccgcccgcga 9360
ccggggtcgt gaagccgatc tgtaccaggg gcagccggcc ggcgtcgggc tccaccccga 9420
gtgcgtccag gatcagcttg aacggcagac tcgcgtggtg caggccgtcc cgccgggcgt 9480
cgcgcacccg ggccaccacg gcggcgaagc tgtccgcctc gtccagttcg acccgcagcg 9540
gcagcaggtt tccgatgtca ccgacgagtc ggctctcgtc gtacgggccc gcgacgtcct 9600
ggggaagtcc gatggtgagg tcgccccggg aggtgtggcg ggcgatcacg accaggaaag 9660
cggccagcag gacgtcccgc cgttcggcgc cgctcccggt cgccaagtcg tcggcggccg 9720
ccgccagtcc gggcggcagc ggccgccgga cgacgtcgct gctcaggtcg atcagccgcg 9780
gacgggggag gtcggtgggc agctcggtgc tctcccagcc cgcgagggcg tcacgccagt 9840
aggtggtcgc ctcgcgggcg gcgtcctgct cgcaccaggc ccgctgccag gccgcgtagg 9900
taccgaactc caccggatcc tgcgcccagg ccggcgctgc gcccgccgcc cgggcgcggt 9960
aggcgtgggc cagatcgccc tgcagcacgc gtgcggagcg cgggtcgagg acgatggagt 10020
ggctggtcag caccagcagg tgcaggtcgg gggcggaccg cagcagccgg aaacggacca 10080
tcgggccgga gttgaggtcg aatccccggc gcagctcgcc ctgcaccacc tcgtcggcgc 10140
gcgggtgcaa ccgctcggcg ggaacgtcct cggtctccag acggatgcgg ggcaccggct 10200
tcagccgctg ggtgccccgg gtgtcctcgt cggagaacgt cgagcgcagc acggtgtggc 10260
gctgcagcag atcggccagc gccgcgccca gcgcttccgt gtcgagccgg ccctcgatcg 10320
ggagggcgaa gccgagcccg taggccaggc tgctgggggc gaaccggtcg aacagccaca 10380
gctgctcctg ccggtaggac aggggggcgg accgggctcc ggcggacaca tcctccagct 10440
gagccaacag cgcgtcccgg cgctcccccg ggagccggcg cagtcggcga agcagctcgc 10500
tctgttgact catgccactg tcctccaagt cggcatgtac accacgacac atacttcaaa 10560
ggctcatcca acaggcaaca ttggagtcaa cgcaatactt caaatgccgc ttagagctgc 10620
gaacaaaagt gggttccgat cagcgggcgg tgtccggtgc gtgcgatcgc aaggcgccgg 10680
aaggtccttg tagcggagct accagggcct ttcggcaacg ccgcgagcgt gcgtgccgga 10740
caccgcgagc cgggcgcccc ttttgttcgc agctcttact ggtcccggcg gcgggtcacg 10800
gcgcgaggga gccggagccg agggtgagac ggtccgccag tacccggacc agcgcggacg 10860
gcgcgtccag gaaggtgtag tgggtccccg gcaggaccac cttgtccgtg ggcgcccagg 10920
cctgccagcc gcccatcaga cggtgcggta cctcgacatc gtcgtcccag cccagcgcgg 10980
tgatggggca gggcaccgta ccggccggac ggaagtactt ccggttggcc tccagatcgg 11040
cgtgcatcac gtcgatgaac aggtcgacga gctcgtcggc ggaggcggcg gcgcccatcg 11100
cggtcagcag cccggcgatc tcgtcccgca gaccctggtc gtccagcccg aggaaacgac 11160
cccagggacc gtcctgcgga gccacctgac tcgacacgaa ataccgcacc ggcgcgggca 11220
gcccgcgctc cgcgatcccg atcaccgtct cgaacgccgg cagggcgccc ccgcagtgcc 11280
cgaacagcgc gtagggccgg gaggtcagca gctccccgac gtcgtccagc agcgcggcgg 11340
ccagctcctc gtaggtgccg aagtgcggct cgcgcatccg gttctcccgc cagggcagtt 11400
gcagcgggac cacctccgcc tcaccgatgg tggcgggcca ggcccggtac atggaggcgc 11460
cgcacccgga gtacggcatg cacagcagca gcatccgggc gtcgggcgac ggctcgcgca 11520
ggaaccagcg ccgctgacgg ggctccatca tgccttcacc cccgccacca caccgttcgc 11580
ccccggtgag gcgaaccgga ccgtgcgcac ctcggagcag cccgcgtcgc gcagccagtc 11640
ggtgtactcg gcgccggtgt agttccgtcc ccaggtgccc accagcatgt tcatgctcat 11700
cagcgccgcg tccaccgggc cggatttgtc gtcgtccacc agcagctcac tgatcaggac 11760
ggtgccgccg gagggcagcg cgtccaggca ggaggcgagg atcttccggt tctggctctc 11820
gtcccagtcg tgcaggatca tcgacagcag gacggcgtcg tgcccggagg gcagcggatc 11880
cgcgaagaag tcggccgcgg ccgtgtcgat gcgtccggtc ataccggccg cggcgatgcg 11940
ttcgccggcg agtgcgcaca cgtgcggcag gtcgacgacg gtgccgctca ggtgcgggtg 12000
ctgccggcac agttcgatca gcgtggcgcc cgcgccgccg ccgacgtcca tgatccggcg 12060
gacaccggag aggtccaccg acgccgccag ctggcgggcg gtcagggtgg agaaggtgaa 12120
gagcccctcc cagaaggact ccggccggtt gccctcctcg aacatgttcc ggtccgggtc 12180
gggcaccggc ttggtggggg cgtcggtgcg taccgcctcg gtcgcccgaa gccaccccgg 12240
cgcggtgtac cgggccacca tctgcacgta accgccgaag tagtcgggca ggccgggcac 12300
caggtagtgc gccgcgacgg cggtgttccg atagcgcccg tcccgctgct ccagcagccc 12360
gagcgcggtg cacgcggtca gcaggatctc ggccggccgc ttgccgatgc cgagccgttg 12420
cgccgtctcg ggagcgtcgg cgtcgccggc ggccagggct tcgaagaggc ctgtctcgat 12480
cgccgcggtc agggtctggg tcttccacag ccccgtcgcc aggtcggtca gcagggtcgg 12540
aggcggggtg ccgggaggcg tcccggagcc ggaaccggcg gccatgtcag tgctcctctc 12600
cggtgcggaa ccgcaccgac acccggtcga acatgatgtg ccgaccgttc tcgaaatgca 12660
ggtcgctcgg ctgctccagg tcgaacagcg gcagccggga cagttcgtcg agcacggcgg 12720
cgaactcggc gcgggccagg tgcaccccca tgcagtgatg gatgccgtgc ccgaagctga 12780
ggtgctgatg ggagttgccg cggtcgggat cgaaggtgtc gggatccggg aacttctccg 12840
ggtcccggtt ggcagcgtcg aacaggagca ccacccgggc acccgccgga atgacggtgc 12900
ccgcgacggt gacctcacgg gtggtgaccc gtgtgaaccg gtcggtggga caccagcggc 12960
gcagggcttc ctccgcgacg gccgggcgca gctccggctc gcggcgcagc ctcagttgga 13020
gtgcgggatc ctgggccagc gtgtgcacga ggcgggtgat cgagttcatc gtgttgtcgt 13080
gcccggcgac gcagaagctg acgatgatcg agtacagctc ctcgtccggc aacggcgaac 13140
cgtcccgccg catggccgcg agccgggcga gatagctgcc gtccggctcg tcgcgcaccc 13200
gctcgacctc ctcggccagc agggcgtgga aggcgggcca gaacttgctc gcgtcccggt 13260
cggccgagat gtaggtccac atcgtccgcg tcgcctcgcg gatccgggcg cggcagctct 13320
cggaaagccc gaaggccact ccgagcacct cgatggcgaa cggctccgcg aactcggcca 13380
tcaggtcccc gtggccgcgc ccggccagcg gcgcgagcag ccgcgcgacc atgcggcgca 13440
gctcaggcac catcgcggag acggcgtcgc ccgtcatcgg agcggccatc cgctcacgta 13500
tcccgcgctg cgccgggggg tcgtagtcga tgggtgagaa catcggcatc ccctcggcgc 13560
gcgggaagtg cacgccgcgg gccgagctgt aggtacgcca gtccgtcagg gcggcgcgca 13620
cgtccgcgta ccgggacagt atccagatgc cgccgtacgc cggtgaccac gtcacccccc 13680
gctcacggat ctccgtgttg atccgctccg cggtccgcgg ggtgtatccg gggtcgtggt 13740
ggtcgaaagc ggtcaccagc cggtcggtgg cgtccgcggc ttcggtggtc atgcgggcct 13800
ccctgccgat gtcttcggtc ggacgaggtg tctcgtgcgg agtgccgggg cggcgccgcc 13860
ggcggacggt gctcagcggc gccggggcac gatgggcggc agcgcggccg cgccggtgcg 13920
cccggcggtg gcccccggga tgcgtccggc cagttcgacg acggtcgggc cggccatcag 13980
gtccggcagt tcgatccgga cgccgaagcg ctgctcgatc ctgctctgca accgggccag 14040
cagcagggag tcgccgccga gttccacgaa gtggtcgtcg gctccgaccc ggtccacctg 14100
cagcagatcc gcgaacaggc gggccagctc ggtctgcgct ccgggtgcgg gtgcccgcca 14160
ggggcgcgac ggacgtcccc gggaccagtc ggccgggggc agcctgcggc ggtcgatctt 14220
gcgggacgtg ttcagcggga actcgtccag ttgcacgtac gcgaccggga ccatccagtc 14280
cggcagccgt tccaccgcgt ggtcgcgcag ctccagcggc gcgatctcgc gtgccgcgac 14340
gaagcaggcg acgaggcggc gctggtcgcc gtcccaggtc gccaccacgg cgacctggtc 14400
gacgtccggg tgctgcccga gcgcccactc gacatccccg agctccaccc ggtggccgcg 14460
tatcttcacg atgttgtcga tccggccgtg gtgctccagg atcccgtcgg gacggaacct 14520
gcccgcgtcg cccgagcggt agatgcggct gcccgcggcg ccgaacgggt tgggccggaa 14580
ccggtccgcg gtctgcgccg gatcgcccag atacccgaga ctgaggcagt ccgagtcgat 14640
ccacacctcg ccgatcccgt cgacgaccgg tgtcaggccg tcgtcctcca gcaggtggac 14700
ggtgaccccg gcgatcggcc ggccgacagg cggggcgctg gcgcggtcgg cgtcggtggc 14760
cgtggcggga gtgcacctct gggaccaggg atggcacgag ttggctatct ggaaggcttc 14820
caggctgccg taggagacgg cgacctcgaa cggcaggtcc gcgggccccc accggttcag 14880
ccggtcaccg cccagggtga ggagccgcag cgcgcactcg cgcggccagg gcagattctg 14940
caggacctcg ccgacggggg tgatcacgaa actctgggtg atccgctcgg cgagcagcca 15000
gtcccgcagg gcgggcggtg acgcgatcac ctcgtggtcg ccgatcacga gctgggcccc 15060
ctgccagagc agaccgccga cctcgtgcag cgcgatcccg gcgccggccg cgccgagcca 15120
ggaaccgcgg tcggcggcgg tggtggcgtt caccgcggcg ctgcgggcgg cgctgagcgc 15180
cagccaggag tgctgggcca gcacgatctt ggcctcaccg gaggaaccgg aggtgaacat 15240
cgcgtacgcg aggtcctcgc accgtggttg cgcacgcggg gcgagtccgc cgccgggtgt 15300
ctctgcgggg agcggagcgc tcgcgtcgag ggtgaccacg ggcagcccgg cggggagtgc 15360
gggcaggggg ttgcccgtac cacggccggg aaccgcggtc acgacggccg cggggcgcgc 15420
cgcggcgacc gcccggtcga accgttccgc gggtacggcc gggtccagcg gcatgcacac 15480
accgcccgtc ttcaggcagc cgagcatgcc gaccacccac tcgatggagc ggcccaggta 15540
catggcgacc atgtcgccat gtcgtactcc cgactcgacc agctccgagg cgagttgccc 15600
ggcacgccgg tccaggtccc ggtaggtcag ctcccgttcg ccgtgccgga ccgcgggggc 15660
atcgggggtg tcgcgggcga acagctcgat acgggagacc accgtggtgg ccggcccggc 15720
cccgggtacg gccggagcgt cctgctggcg gatcatggag tcctcttctc tgaaggggga 15780
tgagccgtgc agggcggtcc ggccgtacgg gtcggtcgtc gggtcgccgg accgtcggtc 15840
agccgacgac cagtcgcatg tcctccggta cgtcgtcggt cagccgggtc accgaaccgg 15900
ccaggatcca gagcacggca cacagggcga gcgcggaggc caggtagaag gcggcgctgt 15960
tgccggccgt cacggcgacg gcactgccca gcagcgggcc gacggcgttg gcgccccaga 16020
cgagcggcag ggagacgccg ccgacgcggc ccatcgcctg cggcggcgtg gccttctgga 16080
tgagggtcat gaccccgacg ttccccacgg gtgtggtgaa cgacgagatg ccctgggaga 16140
cgatcagcac ggccacggcg aacccggtcc cgccggcctg ggacacgggc agcagggcga 16200
acgccaccac cagcatcgag tgggtcatca gcagggtgcg gccgatgccc agccgctcgt 16260
tgatccggcg ggacaggatc gcgccgacca ccaccggggc gccaccgaca ccgagggtca 16320
gcccgacctg cgccgcgctc agtcccaggt cgtcgtaccc gaacacgatg atcatcgcga 16380
ggaccatgga cacggcggtg ttgccgaagg tggccatcag cgtcagatac cgcagcagtc 16440
tgttgccgaa ggtcatggtg atgccaccac gcaggtcggc cagcgtggcg ctcagaccgc 16500
gcgcggtctg ctgcttctcg ggctcacgga accgtacgaa acccagcgcg atggccgcca 16560
gcaggtagga gacggcgtcg acggtgatcg cccgcgcggc gccgagtgcg ctgaccagcg 16620
cgccgccacc gctgcgtccg aagatcaggg cgaccgagcg gctcagctgc gtcttggcgt 16680
tgccgtccac caggtcctcg cgcgccagca gctgcggaag gtaggcctgg taggcgagct 16740
ggaagacgac gtcgaagacg cccttgacga gcgcggccgc gtacaggtgg accatcgtca 16800
ggttgcccag cgcccaggcc agcggcagtg agccgaggac gacgaagcgg accgcgttgg 16860
acagctgcat cagcatccgc cggttcaggc ggtcggtgaa cacccccatg agcggcgcga 16920
gcacgacgaa cggaacccat tgggcggcga tgagcatgcc gaccccgaac gcgccctcgc 16980
cgaaggtgat gacggcgagc agcggcagcg cgaccacgct gatctcgctg ccgagcaggc 17040
tgatcgactc gccgccccag accagagcga agtcgcggtt gcggaacagg acggagcgcg 17100
gcttcgtgct cgtgccaccc gtgggggcgg acgtgtcgcc ggacggttcc ggcggcgatg 17160
tgggcgagac ctctgccccg gcactttctg acgctgccgg agtggtcccc gtcccggatg 17220
acgagttggt cgaggtcatg ccctgtggct cctcattgca ggaattcggt gcgagacgcc 17280
gtgtccggtc gggtgctggc agacgaactc cctcaacgca caccacaatc gacgacggcg 17340
tcacatgcgg ccaggctgaa ccgtacattc agtctgcatc gggcatccag tccttccgca 17400
gagtcggcac gtatgacgag atcacccgac cggatcggat gggcaagtgg cttgaagaca 17460
accctccgcc cccggcgcgc ccctgggcaa gtgtctgcgc gtcacgatat cgtatgtcca 17520
tgagagacta cgaagactac atgcccctag cggtcatcca ggagcacttc tggtcctcgg 17580
acggatcggc aacggaacag gccccgctga ccgagtgcgt ggcgctggcg gtgcggggcc 17640
cactggacgc ggccgctctg cgcacggcgg tcggcgccct gctgtcccgg cacgagatcc 17700
tgcgcagcgc ggtacggctg cacgacggac agccgagcca agtggtgctc cccgtgcccg 17760
agcggctgcc gttgacgatc accactctcc cctccccgga ccgggacgcg gacgcacgtc 17820
aggcccggga ccgggaactg tcccggttcg ccgcgaccgg gatcgacccc gccgccggat 17880
gcgggatcgc gttcctgctg ctgcgcgaca gcgacccggc gggcgaggac accctggccc 17940
tggccgtgca ccacatcttc gcggacgcca ccgcggtccg gctcctgctg tcggagctgg 18000
ccgccgacta cgacgccgcg ctcgccgggg ccccctcacc ggtacccgaa cccgaactcc 18060
agtacggcga cttcacccag tgggaacatg cctcgctgct cccggccgtc caggaacccg 18120
acaccgcgtg gtggcgggag accctgcgtg acgcgcccac cgccctggac gtcagaccgg 18180
accgcccgcg ccgccgtgtc cgccgaggcg ccggccgacg cgtcggcctc gtcctgcggg 18240
gagtcgacgg acccgccgtg cggaacaccg cacggaccct gcacgcctcg ccctacgccc 18300
tgtgcctggc gggctgggcg gccatgatca cccgctccac cggtgacacg gacctgatac 18360
tcggcgtact cgcggccaac cggaccgttc cccagctgga gccgcttgtc gggcagttcg 18420
ccaacaccgt cccgctgcgg ctggacctgg cgggcgcacc ggacctcgcc gacgcggtgt 18480
cccgctgctc ggccgcggtg gccgccgccg tcgagcacgg caggctgccg ttcaaccgga 18540
tcgtgcgggc cgcggccgcc ccgcgcccga ccgaccggcc cccgctgatc cagcacatgt 18600
tcatgccgcg cgtggacgcc gtcggcgaac tgaccctggg gggacagccg gtacggaccc 18660
tggacgtcga acgcgaccgc ggccgcttcg acaccgtcgc cgagatcgac gtcaccggcg 18720
accaggtccg gttgtggctc gagtacgaca gcgacctcta caccccggac gggatacacg 18780
ctctggtgga cgactacgcg ggggtgctgc ggcagtggct ggcccggccc gcgctgcgcc 18840
tgccggaact ggaactgagc gcaccggagc cggccggtct cgccttggac gaccgggagc 18900
cggccgacgg gaagccggcg gaggaactcc gggtggacct gcccggcggc ggagccgtca 18960
cgttcctgct cgaccgctca cccggacccg agtaccggcc cgtcctggcc ggactgcgcg 19020
gggcccgggc ggaggacctg gagctgcggc ccggaggatc cgtgcatcac cccacggacc 19080
tgctgctcgc cggggtcccc accggcctca cggcccggcg cgatccacgg ggccggctgg 19140
agatcgtggt ggagacaccc gccccggtgg cggacgcccg tccgggcacg ggccaggacc 19200
ggctcctcga cctcgtcacg gagatctggg cacaggcact cgaggtgccc ggactcgccc 19260
cggacgacga cttcttcacc tgcggcgggc actcgctgct ggccaccgcg ctggtgagcg 19320
agatgcagga gacactcggc gtcagggtac gggtccgcgc gctgttcgag aatccgaccc 19380
cggcagagct gaccgcccac ctgcgggaga ccgaacccga actggacggc atgctcgaac 19440
tcctcgccgc gctgccggac accacgccgg agaagagcgc cgacgcgccg gagcagcacg 19500
acgcgcaccc cgccaccgcg cagccggcgg gagaaaccgc gcagaccgtg gacacacctc 19560
tgctgtccgg ccagcgtcag ctgtggctcg cccagcaggc cgacccggac gcgctcaccc 19620
acaccatccc gctcatcctg gacctggccg gaccgctgaa cccgggggcg ttcgccgcgg 19680
cgctgaacga cgtggtcgcg caccagccgg gcctgcgcgc cacgttcgtc gaggtcgacg 19740
gcctgccggt gcagcggatc agtccccaca ccccggtcga cgtccccgtg atcgacctga 19800
ccgaccacga cgacgccgcc cgcgaagccg aactgcggcg cctggaacag gagatcgcct 19860
acaccggatt cgacctcacc cgcggcccgc tgctgcgggc ccggatcatc ctgctggggg 19920
cggaccgcgg ccaggtccag ctcctgttcc accacctggt caccgacgag gtgtcccaga 19980
ccctgctgat gcaggagctg agcaccgcct accgggccag gaccgccgga cgcgctccgg 20040
tcctgccgcc gcacgagacc gacctcgcca ccctggccaa ggacgagcag gactccctcg 20100
ccggcgcgga gggcgaacgc ctgcggcggt tctgggtccg tgagctgacc ggagcggccc 20160
cgctgcgcct gcccaccgac cgcccgcgcg gcgaccgggc cggcttccgc ggcgagttcc 20220
tggaacgacc ggcctccgcc gacgcggcca ccactctgcg tgaactggcc ggcgtctgcc 20280
gcacgacccc gttcaccgtg ttctgcgccg ccgtcgcgat cgtgctgaga cggctgagcg 20340
gccagagcga catcgtcatc gggatcccca ccgccaaccg cacgcagcgg ggagccgacc 20400
ggctgatcgg ctgcttcctc aacgtcgtcc cggtccggct cgacctgtcg ggcaaccccc 20460
gcttcgacga actcgtccag cgggtgagcg aggccgtcct gcgctcctac gaacaccagc 20520
agctgccctt cgcggagatc gtccaggcgg tccggccccg gcgagtgccc ggtacccacc 20580
ccatctacca ggtcacctgc gaactgcagc tggagtcctg gatgcccgcc cggttcgcgg 20640
acctggactg cgactaccgc ttcgtctccc acggcacggc ccgctacgac atggccttcc 20700
acgggctgct gcgcccctcc ggactctcgg cgatggtgga gctcgacacc gacctgtggg 20760
accgggcgac cggctaccgg cggatcgacc aggtgctgga gctgctcgcc gtggcggcgc 20820
gcggcccgcg gaccccgatc gacgacctgc ccgtgacgga gcgggagccg cggtccctca 20880
gccggtgacg gccgcgccga acgcgggacc cgccatgccc gcggcgagcg tcgtcccgtc 20940
ggccgggtcg atcagcagga aggagccggt gcgccgcgag tcggcgtagg cgtccacggc 21000
gagcggctcc gcggtacgga tccggacccg gccgatgtca ttggcgacca gctgcccggg 21060
ctccgggtgc gcgacgatcc cgtccaggac gagccgggac cggatctccg cgacgatggc 21120
tctgacggta cgggtggtgt gtttgaggag cacccgctgc ccgaccgcga gcggcgcgtc 21180
cgcgacgtgg cagaccgtcg cctcgacgtc ccggcgggcg accgggccgt cccctgacgg 21240
caccaggagg tcaccgcgcg cgacgtcgat gtcgtcctcc agcaggagcg tcaccgactg 21300
cggcgcccgc gcctcgccga ccgcttcgcc cagcacgtcg ataccgctga tgcgcgaggt 21360
ggcgccggag ggcaggaccg tgacctcctg gccgacgcgg aacgcaccgg cggcgatctg 21420
acccgcgtag ccgcggtagt ccgggtgctc ggcggtccgc ggcctgatca cgtactggac 21480
ggggaggcgg gcgtggcagc cggtcaggtc gtggctgacc gggaccgtct ccaggtgctc 21540
caggacggtg ggaccgccgt accagtccat gttcgccgac ggttccacga cgttgtcgcc 21600
gacgagcgcg gagatcggga tcgcggtgac ctccgggacg cccagctccg tcgcgtacgc 21660
ggtgaactcc tcggcgatcc gggcgaacac ccgcgcctcg tagcccgcca ggtccatctt 21720
gttgacggcg aggacgaggt gcgggacgcg cagcagggcg gcgatggtgg cgtgcctgcg 21780
ggtctgctcg accacgccgt ggcgggcgtc gacgaggacg agggccagct cggccgtgga 21840
cgcgcccgtc accatgttcc gcgtgtactg cacgtgcccg ggggtgtccg ccaggatgaa 21900
ccggcgccgg gcggtggcga agtagcggta ggcgacgtcg atggtgatgc cctgctcccg 21960
ctccgcgcgc aggccgtcgg tgagcagcgc caggtcgggg gcgtcctggc cgcggtggcg 22020
ggaggcgcgc tcgacggcct cccactggtc ggtgaggacc gacttggagt cgtacagaag 22080
acggccgacg agggtggatt tgccgtcgtc gaccgacccg gccgtcgcga aacgcagcag 22140
tgcggtggcc gacggaccgg cccaccgtcc ggtggatcca ctcatcagaa gtacccctca 22200
cgcttgcggt cctccatcgc ggcctcggac agccggtcgt cggcccgggt ggcgccccgc 22260
tcggtgagcc gggacgtggc gatctcggcg acgacctggt ccagcgtcac cgcgtcggag 22320
tcgacggcgc cggtgcagga catgtcgccg accgtgcggt agcggaccag ccgggtctcg 22380
acgcgttcgc cgtccttggg gccgccccac tcgccggcgg tcagccacat gccgttgcgg 22440
cggaacaccg ggcgctcgtg ggcgtagtag atcccgggga gttcgatgtc ctcgcgggcg 22500
atgtactgcc acacgtccag ctcggtccag ttggacagcg ggaagacgcg gacgtgctcg 22560
ccggggccgt gccgcccgtt gtacagctgc cacagttccg ggcgctggcg gcgcgggtcc 22620
cactgcgaga actcgtcgcg gagactgaac acgcgctcct tggcgcgggc cttctcctcg 22680
tcgcggcgcc cgccgccgag gaccgcgtcg aaccgctcgc tgtggatctt ctccgtgagc 22740
ggcacggtct gcagcgggtt gcgggtgccg tccgggcgct ccttgagcac cccgcggtcg 22800
atgtagtcct gcacggaggc gacgtgcagc cgcagtccgt ggcgggccac cgcgcggtcg 22860
cggtaggcga ggacctcggg gaagttgtgc ccggtgtcga cgtgcagcag cgagaagggg 22920
accggcgcgg gcgcgaacgc cttgagcgcc aggtgcagca tgacgatcga gtccttgccg 22980
ccggagaaca ggatcaccgg cttctcgaac tcgcccgcca ctcccggaag atgtgcacgg 23040
cctcggactc cagcgcgtcc agatggctga tcccgtagcg cgggagcggt gtcgtcgccg 23100
tcatgccacg aaccgcttcc gcacgagcag gtccatgagc agggaggccg acgcgtcggc 23160
cgactgccgg accgtcggga tgacgaggtc gggcttctcg ggcgcctggt agggatcgtc 23220
gaccccggtg agaccggtca gctcgccggc ccgctgccgg gcgtacaggc ccttgacgtc 23280
ccgttgcgca cagacctcca cgggcgtggc cacatggact tccaggaatg cggtcccgct 23340
cgcggtgtgc cgctgccgga cggcgtcgcg gctgtcggcg tacggcgcga ccaccggcac 23400
gaggacgatc acgccgttgc gggcgaggac ctcggccatc accccgatcc gctgcaggtt 23460
ctcgtgccgg tccgcccggc cgaatccgag ccctgccgac agtgtcttcc gcatgacgtc 23520
accgtccagg acctccactc ggtgtcccgc gtcgcgcagt cgcccggcga ccaggaacgc 23580
gaccgtcgtc ttgcccgcac tcggcagccc cgtcagcagg acggtcacgc cgcagtcaca 23640
tgattcaggc ccgatcacca tggtcgatcc gaaccgcctt ccaggtacag taggaaaaca 23700
cgacaaaagc tgaaatactc gatacatgac gacaactgac aagaccgaca gccgacttgg 23760
gccctacgtc gagcacctgg gcctgcagtt cgagcggatc gacccggacc gggcggtggc 23820
gtactggtcg gagcgcgccg acctgctgca gcctcacggg atactgcacg gcggggtgca 23880
ctgcgctgtg gtcgagtccg tcgccagcgc ggcagcggac cgctggctgg gcgaccgggg 23940
gaccgtggtc ggggtcagca acagcacgga cttcttcgct cccgcgaccg tggccgacgg 24000
ccgtctcacg tcgacggccc tgccggtcca ccgcggcgcc acccaacagg tctggtcggt 24060
ggagacggtg gacgcggcgg gccggctcgt cgcccgtgga caggtccggc tgcacaacct 24120
ccgctgactc cggcatcacg gacggacagg aagggggacg gccccagagc tccggaacag 24180
aagcaggcca cggcgagggg cgcccggcgc gccctcgccg cggcgccgga aggccctggc 24240
aggtgcgcca caaggacctt ccggcgccga gcgatcgcac gcaccggaca ccgtccgctg 24300
accggatccc gcttctgtcc gcggctctca gccctgccag ctgtgcggcg cccggaactg 24360
cggaatgcgc agcgggcgcc gccggccggc gggggagtgg tcccgggagg ggacaccgcc 24420
cccggcctgc cgccgggccg cggcgcagac cagcagcagg gcggtgagag ccgcgagttc 24480
ctcgggctcc gcgtgtcccc tctcgacccg gagcaaccag gcggggccgg tctgccggga 24540
cagggctcag ccccgttccg gggcggcggt gagcacgccc tcggtccggg ccgcgcgggg 24600
ccggctccac cgcctcatga ccggccgctc caccagcgtg tacagcagcc acgccagcac 24660
caggctggag gcgatggtca gacccgccag gccgaggatc tccggccatg tcgagggctc 24720
cggccgagcc ccccaggcgg gggagctgcg gccgatccag ccgtaggcca ccaccagcca 24780
gtgcacgagg tagagggcga aggagatctc gcccaggaac accaggggcc tgctgccgag 24840
ccaggtccgg ccgttgcggc cgttgatgtc ggcgctggcg acggccgcga cgaggaaggc 24900
gagcggcagc gacgtgagcg ccacgaaccc gagctcgtcg ggcatgctgc cctcggcgac 24960
gagcggaatc agggggaaca cgagcgacac ggcgaacggc agccggatcc accgccggtt 25020
gacgacgatc agcgccgtca cgatgcccag cacgaactcc agcatccgca ccggcgggaa 25080
gaacgtcatg aaccacatgg agagccagtt ctggccggtg cccgggtgga cacccccgga 25140
cgtcggcatc caggaagcca ccagcggcac cgcccagatc gccgcgtaga ccaccgcgaa 25200
cgtcacccac aggcgggccg gcgggatctt ccgcagcagc ttcaggagcc acgggaacgc 25260
caggtagaag agcacttcgc aggcgagcga ccaggtcgga gtgttcgacc acaggttgta 25320
ctggagcttc atgtccggga tccaggactg caccagcagc agtgtcgtgc tcgcgtcgcg 25380
tcccgacagg gcatggcccg cgcccgccgc gagcagaagg gcggcgagca gcgtgaccag 25440
atggttcgga tagatcttga agaagcggcg ccgccagaac cgccccacgg agtcctcgtc 25500
ccgggcgacc caggtcagca cgaacccgct caggatgaag aagaacgaca ccccgaaggc 25560
accgcactcg tacaggaccc tgatcgcgaa cgtctgatcc ggagccaccc gcggcagcag 25620
gttggtcccg acgtgcgaga tcaccaccag gaacgcggcg aagaaccgca gcccggtcag 25680
ggacggaagc cggtggaccc ggtgatcggg cgtggcgacg gacagctgtg gtgccgccgc 25740
cggtctctgg atggtcatgg ggcctccctg cgggacgggt cgagctcgag ggccagcgcg 25800
cgcaccacgg tggagcggta gaagttccgt acggggatgc gcagcccccg ttcccgctcg 25860
atcaggtcga tcacttccat ggcggccagc gaggttccgc cgagcacgaa gaagtcgtcg 25920
tcgggggccg cgtcctgttc cagacggagt acgcgtcgga acacgctctg cacgtaacgg 25980
atgcggtctt cgtcgtggac cagcccaggg ctcgccatcg tcatcccctc atggttgggc 26040
cggcggcagt gtgctgcgcc ggtgattcgg ctcgtgctgc gtgcaggtgc cgggccgcgt 26100
ccgccaccgc gccggtgctc acgccgagct gctccagcgc ggcccgctgg ccggcgacgc 26160
gcacgccggg acggcctcgt acggccacgc ggcgcaccgg gagagccagt tcggcggcga 26220
ccacctcggt gaccgcgtcg cccagcccgc cttgcggccg gtgctcctcc accgtcacca 26280
ctccggcgcg gccccggcac gccgccaaca ggccctcgtg gtccaacggg tgcagggtgt 26340
gcaggtccag gacctgcgcc tcggtgccgt cggcagccag ttctgccgcg gcctccagcg 26400
agaggagcag gggcagcggt cccgcggcca ccagcgtgac gtcggcgccc tcgcgcagga 26460
cccgggcgcg gcccagccgg aaggcggggg caggccccgg cggcacgggg gtggccgagc 26520
ggcccagccg caggtaggcg ggaccgggca cctcgtgcaa ctggcgcatg gcagggccga 26580
tctggcgtgc gtcaccgggt acgacgacgg tcaggtcgga cagggcgcgg gtcaccgcga 26640
ggtcctccag cgcgaagtgg ctggtgccga agtgggcggc ggagagaccg ccgtgggtgg 26700
ccaccacccg tacgggtagc cggtttccca cgacgtccag cttgaggaac tcgcccgcgc 26760
gcatcgtggc gaaggtggcc atcgtgtgca cgtagggaag gaatccgcgg cgggcgagtc 26820
cggcggccac cgtcatcagg ttcgcctccg cgatcccgac attgacgtac cggtccggga 26880
agcgctcgcc gaaggtgttc tccaggccgc cggtgtcgga gtcgagacag accacgcggc 26940
tgtcggcggc ggccagttcc agcagcgcct cgcggtacgc ggcccgggga tccgggccga 27000
ggaccggctg cggggtctgc tcggcggggc gcgccgtcgt cacgccgtac ctccctgcac 27060
ggggtggccg agcgcggcca gggccctgct gaccagggcc ggtttgagct tggcgtagtg 27120
accggccacg cgggcctcga ggaacggcac cccgcggccc ttcacggtgc gcgcgagcac 27180
cgcgaccgga ccgcccgggg acggctgcag ggcctcgtac agcgcgggca ggtcgtgtcc 27240
gtcgacggtg cgtgtctccc agccgaacgc cgcgaacttg gcgtccagcg gttcgagtcc 27300
gacgcagtcc tccgtaccgc ccgtgatctg cagcccgttg cggtccacca ccgcgaccag 27360
cccgggcggc ctgcggtgac cggcgagcag ggcggcctcc cagacactgc cctcctgcag 27420
ttcaccgtct ccgagcaggg tgtacacgcg tccgtcgccg ccgtgcagct gctcggcgat 27480
ggccaggccc accgacaggc cgagtccgtg gcccagggaa ccggtggcga actcgactcc 27540
gggcaggtcg tgcgaggggt ggccgaacag ccggctgccc ggcgcggtgt agctgtcgag 27600
ctcgtccgcc gggagccttc cgagctggac cagcgtgctg tacagcgcgg gtgcggcgtg 27660
ccccttgctc agcacgaacg agtcgcccgg acgcaggagc cgggtgtgca ggacggccag 27720
gacgtccgcg cacgacagac tgccgccgac gtgcgcgccc tcgggcccgc ccgccatgcg 27780
gatcacgtcc tggcggatcc gccgggccat gtccgccggg tcgagcggct gcggaggcgg 27840
cgcgggggcg ggtggctgcg gtgggccggt gaggggtacg gccatcgtgc ggattccctt 27900
cggacgggcg tcggtccggg acgtggacca ggacgtggag cggaggaacc ggcgggggag 27960
cggcgggcgg ctcacccctg gcgcagcgcc tcccgccagc cgggagcgga gcgggaggtg 28020
gccatccggc ggacgagttc gaggttctgc cagaccttct cgaacgcgtc ggcgtacgac 28080
tgcagcaggg gcccggcggc cggatcgagg tgccggcgct gcaggcacag cgatccgtcg 28140
acgacggcga cggtcaccgg gaggagggcg tccacctggt ccgcggcgtc cgccggggac 28200
gcacccgggg gacggaacgc gggctgcgcc gcgagcggag cgacctggta ccgggagacg 28260
ggcaccccct cggcccgcag gatccggtgc agggcggcgc gcagggcgca gcggtgctcg 28320
ggcccgaggc cgaacgcctc cgggtccggc atcagccgca ggatgtgcca ggcgtgggtg 28380
cgtccggccg ggacctgtgg cacccgcagt ccgggcagct gagccagccg gttcaggaac 28440
cggcggattc ccggctcgcg gcggcgggcg tagtcgtcga agcgggccag ctgggagcgg 28500
gtgaacgcgg ctgccatcgg tgtgatccgg gcgttgaggg cggcccggtg ggcgatgtag 28560
ctgcgagccc cggtgggttc cagctcgccg aagttgcgca gccgtaccag ctgttcgtgc 28620
aggctgtcgt cgtcggtggt gatcagtccc ccctcaccac agctcggcag gttcttgtgg 28680
ggatgcaggc tgaagcagcc cgccttgccg agggcgccga cccggcgccc gtcgagctgc 28740
gcgccgtgtg cctgggccgc gtcctccacg acggggacgc cccgtcggcc ggccagttcg 28800
tccagcgccc gcatgtcggc cggcagtccg tgcagatgta ccggcaggag cgcggcggtt 28860
cgcggtccgc agaccgctgc ggcggaccgc gggtcgagtg tgtacgtgtc gggatccacg 28920
tcggcgaaca cgggtgtggc gcctgccgcg cacaccgcgt gggcggtcgc gttcatggtc 28980
agtgccggga cgatcacctc gtcgcccgga ccgatcccgc agccgcgcag ggccagatgc 29040
agtgcggcgg tgccggaggc gacagcgaca cagtgccggg tcccggtggc ctgcgcccag 29100
gcccgttcca ggccgccgac ctcgcgttct ccgggggagg cggcgttcag cgggccctcg 29160
cgcaacaccc gcagcacggc ctgttcgtcg gcggtggtga ccagcggcca ggtgccgccc 29220
acgctgtcct tcggtacggc ccggtctccg ccgagcaggg ccaggcgggt cgtggcgtgg 29280
gcgagctcgg gcatggacgg gtacctccgg tcgtacggct gcgagttccc cactgtgcag 29340
agtctgcgta ctcggcgccc cattgttgcc gcatcccgga gagactgtcg acagtctgaa 29400
tcaactgatc tttgggggtg tgcccttgtg gagtctgtgt tagcttctgc agtgttcttg 29460
gagacactca tcacacaatc tacaggaggc aacacatgac tacggcgctg agcgtgccgt 29520
gtcccgagtg cgccgggacg ctggaggtac gccctgatgt gctcgccgcc gagatcgtgg 29580
tctgcggaca gtgttcgagc gagctcgagg tcgtggcgac cgggcccgtg gtgctggccc 29640
tcgctccgga ggtcgaagag gactggggcg agtagtcgtg aaggtgagcg tcgtcggcgc 29700
ggccggatac accggaggtg aactgctgcg gctggtgcat gcccacccca agctggaact 29760
ggcacaggcg ctttcgcggc ggcatgcggg ccggccggtg gcagaggtgc accccaacct 29820
gcatcatctc gccctggcct tccggtcccc cgaggagctg gagccggtgg acgcggcgtt 29880
cctggccctg ccctccggcg cctccgccga ggtggagaca gcggtgacgg gccgggccac 29940
gaccatcgtg gacctgagcc ccgatttccg gggcggccgg ccctcggcgg acggcaccgc 30000
ctaccggacc ggcctgcccg agctgttcgc cggcgaactc gtcggcgcca cccggatcag 30060
cgtgcccgga tgcatggcca ccgcggccgt cctggccctg gctccggtcc agcgcagtgg 30120
tctggtggcc ggcgacgtgg tggtggacgc ccgggccggg tcgtcgggct ccggagcgga 30180
gccgagccct gcctcgcacc acgcggagcg cggcaacgcc ttccgggtct acaagcccgc 30240
gggacaccgg cacgagaagg agatcgccgc actggccggg gtgcgtgccc ggatgacggt 30300
gaccgccgtc cccacggtgc gcggcgcgca ggtgctctgc catgtcacac ccccgcggcc 30360
ggtcacccgc cgcgaggtgc tgcacgtgct gcagcgcgcc taccgagacc acccgttcgt 30420
ccggatcgtg tcccgcgggc gcggggtgca ccggatgccc gatccgcagt tcctcggcgg 30480
caccaatttc gtcgacatcg gttgcgccgt ggacgaggac ggcggccgca tcgtcctggt 30540
cgccgcgctg gacaacctcg tgaagggggc cgcggggggc ggggtgcaga gcctgaacgt 30600
ggcggccggc atgccacagg acgccggact ggaattcacc ggactgcacc ccgcctgatg 30660
ccggcccgtg aacacggccc gtccgaacgg cgcgagggga acagcggcac gagacgaggg 30720
gaggtgagcg gggaacatgg cgcaggatcc gagggggcac gagccgaacc cgctgatcgg 30780
cgggttcgcg cacgacgggc cgatcgtggt caagatcggc ggcgggagcg acccggagcc 30840
cgtactggac gaggtggccg aactggcgct cgccggccgg ccgacggtac tcgtccacgg 30900
cggcggagca gtcgccgacc tgctgtcgga acaactcggc gtggagcggc gggtgatcag 30960
gtccccggac ggcacgcaca gccgtcgcac ggacgcggcc atgctcgatg tcatcaccct 31020
cgccctgctc gggcgggtga aaccacgtct gatcagcgga ttacgggcac gcggcgcacg 31080
gtcggtgggc ctgagcggcg cggacggcgc cctgctgacc gccacccgca aaccggccct 31140
gcgctccgtg caggacggcc ggaccgtact gatccgggac gaccggtccg gccgcatcga 31200
gcgcgtcgac cccgcgccgg tccgcgcggt gctggaccgg gggcacgtcc ccgtggtctc 31260
cccgccggcc agcgacgccg cgggcaacct gctcaacgtg gacgccgacg aggcggcggc 31320
ccggctggcg accgccctgg acgcgtcggc cctggtcctg ctcaccgatg tgggtggcgt 31380
cctggccgac ctcggcgacc ccgccacccg gatcgcgcgg gtgggcccgc accacctgga 31440
aggcgacgtc gtccgaggac ggatgcggca caaggtacgc gccgggctgc gggccagcag 31500
gaccgtgcgg caggtggcca tcggcgccgc gcatctgcac cggccgatcc accaggccct 31560
ctccggtgcc ggcagccggc tgtgcgacga aggggagagg cccgaagatg gctgaccacc 31620
agcgcaccgc cgaccgtacc cgccccgcgg cgggcgacgg ggactacccg gcctggctcc 31680
tgcgggccgc gctcggcatc ccctcggtct ccggccggga actgcccgtc gcccgcttcc 31740
tcgccgaccg gatggccgag ctgggcctcg actcgtacgt cgacgaggtc ggcaacgtgc 31800
acggcctgat cggcccgcgc acgggcccca ccgtcatgct gctcggacac atcgacaccg 31860
tccccggaca ggtaccggtg gtccaggtcg gcgatctgct ctacggccgc ggcgcggtgg 31920
acgcgaaggg ctccatcgtc accatgatct gtgccgcggc ccgcaccgag ggagtacgcg 31980
tgcacgtgat cggcgcggtc ggcgaggagg tcgccggctc ccgcggcgcc caccacgtgc 32040
tggccacggt gccgcccccg gcggcgctcg tgatcggcga accgagcggc tgggacggcg 32100
tctgcctggg ctacaaggga cgtatcggca tcggctacga gatcagccgg ccacccctgc 32160
acaccagcag cccggagccg accgccgtgg agtgcgccgc ccgcttcgcc cacgacatac 32220
aggagtatct gcggggactc tcgcccgaac aggacgaccc ggtcgccttc ggggtcgccg 32280
ccgcgacgct cgtccggctg cacggcgatc tggccgaggc cgaggccttc ctgagctgcc 32340
gggtgccgcc ggggttcgac ttcgccgcgc tggaacgctt cgcccgcagc agaccgcaca 32400
gccgcatccg ggtggacgaa cgggtccccg gagtgacccg gccgcgcacc gaccccgtgg 32460
tgagccagct ccgcgccgcg atagccgccc agggagtccg cccgacgctc aaacgcaagg 32520
ccggcacctc ggacatgaac accctcgccc cctggggcat ccccatggcc gcctacgggc 32580
cgggcgacgc ccacctggac cacaccaccg acgaacacat cccgctcggc gacctgtacc 32640
gcgcgatcga cgtcctggcc ctcgcgctgc cccggctggc gagccgcctc gcggcgccgc 32700
cgcggaccac cggcgcgccc gagctgctgg aggcatgatg atgctcgacg gcgacgtcgc 32760
cctgatcacc ggtgctgctc gcgcgctggg ccggtcgcac gcgctggcct tcgcccgcga 32820
gggcgcggac ctgatcctcc tcgaccgctg cgcccccgac gacggcccct acccgatggc 32880
cggccggcgt gacctggagg agaccgcgga agcctgccga aggctcggca gccgcgtcct 32940
gaccggagtc gccgacgtac gccaccaggc ggacgtggac aaggtggtgg acgccgggct 33000
cgacgcgttc ggccggatcg acgtcctgct caacaacgcg ggcgtcctcg gtcccggcgg 33060
cgtccgcgta cacgaactca gcgaggaaca gtggcagttg gcgctggacg tcaacctgag 33120
cggcacttgg cgcacctgcc gcgcagtgct gccgcacatg gtcgagcggg gcggcggacg 33180
gatcgtcaac accgcctcca ccggcggacg cgtcgccttc gagcggtacg ccaactacgt 33240
cgcctccaag cacggcgtga tcggactgac caagtccatc gcgctggagt acggacgcca 33300
cggcatacgt gccaacgcgg tctgtcccac caccgtggcc gacgacgccg cactgggcac 33360
ccgcagcacc cgcgccgtgg ccgcgtcgat gggcgccgaa ctgcccgact acgagcgcac 33420
gtcggcggcg ctgcatccga tcggccgact ggtcaccgcc gcggaggtgt cggccgcctg 33480
cctctggctg gccggtgacc ggtccggcgg tgtcaccggc accgaactcc tggtggacgg 33540
cggcttcacc gcccactgag ccgcccgccc gcaccacgtc cacgcgaacg acctgccctg 33600
ccgcggaccg ccgacccgac aaggagtcct ctgccatgga tctgcccacc gagcagccac 33660
ggtccctgcc ggacacgccg gtggccgccg cggtgccgcc cgatctcgcc gccctgcgct 33720
cgcggctggc cgaactgagc ccggaaaaac gccggttggt ggagcgactg ctggcccgtc 33780
agggcgccgg cctcggcatc acccgtcgcg ccggcggtgc gcccgaggcc ccctgctcct 33840
tcgagcaaga gcggctgtgg ttcatgtacg agctgctgac ccggcgcgag atctttcacg 33900
tcccggtcgc cctgcgcctc gaaggcgaac tggaccccga cgcgctggag cgggcactgc 33960
ggcagctcgg ccgacggcac gaggcactgc gcaccgtgtt ccggcagcgc gacggccgcc 34020
cctaccaggt ggtacgggaa cagctcgatc tcccgctgga gcgggtcgac tgccgcaccg 34080
ccgccgatcc cgccctcgcc gcccggcagc aggcctccgc actggtcacc gaggacttcg 34140
acctcgaaga aggacccctg gtgcgctgca cgctgtaccg gacgggggaa cacgaacacc 34200
tgctggccgt ggtgcagcac cacatcgtct ccgacaactg gtcgctgggc atcctgctcg 34260
acgacctcgg caagctgtac gcccgcgagc tgggggcacc gggggagctg cccccgctcg 34320
acctgcacta ccccgacttc gccgcatggc agcgggccac ggtcgacagc acgacgatgc 34380
ggcgcaccct cgatcactgg cgcgaccgcc tggacggcgc cccggacagt ctcgacctgc 34440
ccaccgaccg tccccggccc gcggtccgcg gcagccaggg gaagttccac cacgtacggt 34500
tcgacgccga cctggtcgcc gggctgcgcg aagcggcccg gcagcacgac accaccctcc 34560
tgggcgcgtt cctcgcaggg tacatagccc tgctctcccg cctcgtgcgg tcggactcgc 34620
tggtggtcgg tgtaccggtg gccgggcggc cgcgcgcgga gatgcagcag atgatcgggt 34680
acttcctgaa ctggctgccg atccacgtcc aggtcggtga ccggccggac ctgcacaccc 34740
tgatacgccg caccggaacc gccctgagcg aggcgatgac ccatcaggac gtgccgttcg 34800
acatgctggt gcgcgagctg cggccctcgc ggcgccccgg ggagacaccg atcttccaga 34860
cctccttctc gctgcgtgac ggcgccccca ccccgccggc catgccgggc atcgacgtca 34920
ccttcgccga actcgacggc ggagccaccc attacgacct gatggccgaa ctgtggtgcg 34980
agggcgacga ggtcgtgggc tacgtgccct acgacgacga actgctggac gcgcagacgg 35040
tggcccgctg ggccggctgg ctcaagacgc tgctgcgcgc cgggctggcc gcaccggaca 35100
ccccggtggc ggacctggag atgctcgctc ccggcgagtc cgtggtgatc ccggcccgcg 35160
cgccggtgac ggccgagggg ggcgtcgccc ggccggcccg gaccctgcac ggggtgttcg 35220
ccgcgcaggc ggctcggcgg cccgacgccc ccgccgtctc cgacgagcgg gaccggctca 35280
cctacgcgga actgtccgca cgagccgacc ggatcgccgc cgccctgcaa gagcggggtg 35340
agggacccgg cagcatcgtc ggcctcgtcc tcgaccgcac cgtggacctg cccgccgccg 35400
ttctcggcgt gcttcgcgcg ggtgccgcct acctgcccgt cgacccggag aatcccgccg 35460
gacgcaccgc tgaccagttc accgagtgcc gggtccggac cgtgctgacc accccggcca 35520
ccgcgtcggc acccgcgttg gaggaactgg acgacggacc gcgcatcctc gtggtggact 35580
ggcaggaccc cggatggcgg gagcagaccc ccggcccggc tgccgtggac gtacccgcgg 35640
acgcgcccgc ctacgtcatc tacacctcgg gatccaccgg ccgccccaag ggcgtgctcg 35700
tcacgcaccg caacgtgctc cggctgttca ccgcctgcga cgagcatctg agggtggggc 35760
cggacgacgt atggaccctg ttccactcct acgccttcga cttctccgtc tgggagatct 35820
ggggtgcgct gctgcacggc ggcaggctgg tggtcgtccc gcagtgggcg acccgcgccc 35880
ccgacgtctt cgccgaactc gtacgggacg agcgggtcac cgtgctgagc cagaccccgt 35940
ccgcgttcgg ccaggtgagc gagaccctgc tgaagcaccc cgacccggcg gcgctgcggt 36000
acgtggtgtt cggcggcgag gcactcgatc ccaccgcgct gcgtccctgg acgcgtgtgt 36060
acggcgaccg gcggccggaa ctgatcaaca tgtacgggat caccgagacc accgtgcacg 36120
tcaccgcgcg gccgctgcgc gagaaggacc tcgccgggac ggcgtccccg atcggacctc 36180
cgctcgccga cctgtccctg tacctgctgg acgattccct gcgtccggtc ggcaccggtg 36240
tacccgggga gatcttcgtc ggaggggacg gggtgagcct gggctacgtc gccaacccgg 36300
ggctgaccgc ggagcgcatg ctccccgacc cgttcgccgg acgtcccggc gcccgtatgt 36360
accgcagcgg cgacctggcg gtccgccgcg gcgacggaga gctggtctgt ctgggccgcg 36420
ccgacgacca ggtgaagatc cgcgggcacc ggatcgagcc gggtgaggtc cgggcggcgc 36480
tggacgccct ggccgtcgtc gcccgtgccg ccgtggtggt ggaacgcgac cgggtgggcg 36540
ccgccgtcct ggcggcctac gtcgtaccgg ccgaggggaa cgccggcgtc agcggcaccg 36600
ggattcgccg gaccctgctg cgctccctgc cggaatggat ggtgcccgcc tcggtcaccg 36660
tcctcgacga gctgcccctg acccggaacg gcaagctgga ccgccgggcc ctgaccgacc 36720
gcaaggagca cgcggcgccg gccggaccgc gcggcgaggc gccccgcagc gacaccgcgc 36780
ggcaacttgc cgagatctgg caggagctgc tcggcgtccc cgcggtgggc ggcgaggaca 36840
gcttcttcga actcggcggc cactccctga tggtgatgca cctcgtcgcc cggatacgca 36900
cggccttcgg cgtcgagatg ccggtggaga ccctcttccg ccgccctcaa ctgcagccgc 36960
tggccgacga ggtggacgcg gcacgtaccg cggcgcagag ggccgcggcg ccggagccgg 37020
gacaggacgc ggcggccgaa ctcgccgacg tgcgcgccga cctggccgcc cgggccgccg 37080
ggatcccccg cccgggggcc cgggcggacg cggaccgcga cacagtcctg ctcaccgggg 37140
ccacgggctt cgtcggccgg ttcgtgctcg ccgaactcct tgccgcgggg gcccgggtga 37200
tctgcctgct ccgcggcggg accgcccgcc gggaggagct ggtggcgggg atggcggacc 37260
tcgggctgtg gcacgaggag cacgccgcac ggctggaact ggtcgacggc gacatcgccg 37320
aacccgggct cggcctcgcc ggaccggacc gcgaccgcct ggccgaccgg gcgggccgca 37380
tcatccacgc cgcggcctgg gtgaaccacg tctacccgta cgagcggctg gccgcggcga 37440
acacgcactg catggcaggt ctgctggaac tcgcggcccg cggccgccgc tcggctctca 37500
ctgtggtgtc caccagctcg gtggccgact ccgccgcgta cccgcccggc tcgaccgtgc 37560
cgccgggccc gctgaaggca ctgccgtccg ccgcgaacgg atacgtgcgt tccaaggcgg 37620
tggccgagca gtacctccac ctggcagccg aactggacgt acccgccgcg gtgatcagga 37680
ttcccagcgt cttcggcgat cagcggcggt accagatcaa ccccgcagac gccgtgtgga 37740
gttggtgccg ggcgatgatc gagaccagcg gcttccccga gagcttcgcg cagccgggca 37800
acgaactgtt ccaggcgctg cccgccgacg ccgtggcccg ggcggtgctg ctggccgacc 37860
gcgatcacac cgaaccgggc acccgctacc tggacgccgt cccggccgcc gtcggcacga 37920
cagaggacct gctggccgct ctgcgcgccg ccggtcatgc actgagcccc tgcccggacc 37980
acgcgtggta caccgccgtg ggcgagctgg accccggccg ggtctgggtg gcgggcatcg 38040
cagggcaggc cgccgcccgg ctggccgcgg atccgtccgc cacggccccc cgcaccctgc 38100
gccgcttcac cgcccccgtg gagcccggtg agctggacga gctgttgcgc acccgggccc 38160
tgtactcctc ggcccagctg gccgggtaca tccggacgct cgacgcgtcc ttcccgcccg 38220
ccggcaacga ccgggagaaa gcccggcccg cgcgcggctg acccgcagga ccgcacatca 38280
ccgcatgacc gatgggagtg ggagtgagtt cggacgagcg cacgtacctc gtggtgcgca 38340
accacgagga gcagtactcg atctggcccg ccgggcagga cgtcccggcc ggctgggccg 38400
cggagggctt ccaaggcccc aagagcgact gcctggcaca catagccgag gtatggactg 38460
atctgcggcc gctgagcctg cgccgcgccg gtgacgacaa gggagcagag cgcgttggct 38520
gagaatgtgc agaacccgcc cgtggagccc ctcgcggtga tcggcatgag ctgccggttc 38580
gcccccgacc tcgacacccc cggccggctc tgggagttcc tgcgcgccgg gggcagcgcg 38640
gtgggcgaga tgcccgaccg gcgctgggac ccgtacgtga cggacagccg cacccgcgac 38700
atcctgcgca cgaccacccg caagggttcg ttcatgcggg acatcgaggg cttcgacgcc 38760
gagttcttcc agatcacgcc ccgcgaggcc gagtacatcg acccgcagca gcggatcatg 38820
ctcgaactcg cctgggaagc cctgtgcgac gccggactgc cgccgacctc gctggccggc 38880
accgacgcga gcgtctacgt ggcggcgaac tccaacgact acggccgccg cctgctcgaa 38940
gacctggacc gcacgggcgc ctgggccgtg aacggcacca ccttctacgg catcgccaac 39000
cgcatctcgt acttcctgga cgcgcacggc cccagcatgg ccgtggacac cgcctgcgcc 39060
ggatccctga ccgccctgca cgtcgcgggc caggcgctgc acaggggcga gacgagcgtg 39120
gccatcgtcg gcggcatcaa catcatggcc tcgccggcgc tggtcgtggc cctggacgcg 39180
gcctccgcga cctccccgga cggccgcagc aagtcgttcg acaaggcggc cgacggctac 39240
gggcgcggcg agggcggcgg agtcgtcgtg ctcaagcgac tgtccgacgc ggtgcgcgac 39300
ggtgacccgg tgcacgggct cgtcctggcc agcggtgtct tccaggacgg ccgctccgac 39360
ggcatgatgg cccccaacgg cagtgcccag cagcgcatgc tggaagaaat ctaccggcgg 39420
tccggcatcg accccggcac agtgcagtac gtcgaggcgc acggcaccgg cacccagctc 39480
ggtgacgccg cggaggccca ggccatcggc aatgtcttcg gccccggcag ggacggcgac 39540
aaccccctgc tgatcgggac gctgaagccg aacgtgggcc acgtcgaggc agcctccggc 39600
atcgcgggcg tgatcaaggt cctgctgggg atgcgccacg gcgaactgcc gcccagcccg 39660
cacgaggagc ccgatcccgg gctcgggctg gaggccaggg gactgcgtct ggtcgccgag 39720
ccgacgccgt ggccgcgggg cgaacacggc atgcgcgccg gtgtctccag ctacggcgtg 39780
ggcggctcga tcgcccacgc cgtactgcag caggccccgc cgcgcccgga caggaccgag 39840
cgcccggccg ccgccgccac gggccgcccc caggtcttcc cgctgtccgc cgcctccgag 39900
cagggcgtac gaggactggc cggctcggtc gccgcgtggc tgcgggcaca ccccgagacc 39960
gccctggacg atctcgccca caccttcacc gcccgccgct cccatctctc ccgccgggcc 40020
gccgtcgtgg ccggcaccac ggaggaactg ctcggcggac tggacgcgtt ggccggcggc 40080
gagaaatccc ccgcggtggc cctggcatcg gcgtccgggt tcggtgacgg cggcgcggcc 40140
ggacccgcct gggtgttctc cgggcacggc gcgcagtggt ccggcatggg ccgggaactg 40200
ctcaccaccg agcccgtgtt cgcccaggtc atcgacgaac tcgccccggt cttctccgaa 40260
gagctcggat ggaccccgcg cgaggcgatc gaggcgggcg gcccgtggac cgtcgtccgt 40320
acccaggcga tgaccttcgc gatgcaggtc gccctcgccg aggtgtggtc ggacctgggc 40380
ctgcgtccgg gcgcgatcat cgggcactcg gtcggcgaga tcgccgccgc cgcggtggcc 40440
gggagcctgg accgcgcaga ggcggcacgc ttcgcctgcc gccgcgcccg cgccctcgga 40500
aagatcgccg ggcgtggcgc gatggcgatg gtgcccatgg cgttcgccga cgtcgaacag 40560
cgggtcgccg gccgggacgc agtggtcgcg gcgatcgccg ccagccctct gtcgaccgtg 40620
gtgtcgggtg acacggccgc ggtcgaggcc ctgctcgcgg acctggaggc cgacggcatc 40680
caggctcgcc gggtcaacac cgacgtcgcc ttccacagcc cgcacgtcca ggagatcctc 40740
gacgaggtca ggcaggccgc cgccgcactg cgggccggaa caccccgggt gacgctgtac 40800
agcaccgccc tcgcggaccc gcgctccgac gctccccgtg agggcgagta ctgggccacc 40860
aacctggccg acccggtccg cttccaccag gccgtacggg ccgcgctcga cgacggcacc 40920
cgggtcttcc tggaggtctc ctcgcacccg gtggtggccc actccatcac cgagaccgca 40980
ctggacgccg gggtccccga cgcccacgtg gcgatcaccc tgcgcaggga acagccggag 41040
cagcgcaccg tgctggccaa cctggcccgg ctccacagcc tcggcactcc ggtcacctgg 41100
tcctacgacg gtgacctcgt ggacgtcccc gcggtgcgct ggcagcacaa gccgtactgg 41160
atcttcccgg acaccgcacc ggaacagggc gcgggcctgg gccacgaccc gcagacccac 41220
acgctgatcg gcgcgcgcac caccgtcgcc agcgctccgg tccagcgcgt ctggcagacc 41280
gaactgcaca tggagaaccg tccgtacgcg cagtcgcaca aggtcgtcgg cgtggagacg 41340
gtgcccgcgt ccgtggtgct caactcgttc atcaccgccg ccaccaacga gggcgaacgg 41400
gcctgcggac tgcgcgacat cgtcttccgc atcccgctgg ccgcgcaccc gacacgggtg 41460
gtccaggtgg tgctggaaca ggacaaggtg cgcatcgcca gccgcatcaa gcgggaccag 41520
gagagcggcg gcgtccgcga cgacgagtgg ctcacccaca ccaccgcgac cgtcgtgcac 41580
gagccggagg tcggggcgcg tccgatggag gacccggacg tcatccgggc ccgctgcccg 41640
gtgtcctgga cctgggccaa ggtcgacggg atcttccgga ccatgggcgt ggacgggtac 41700
accttcccct gggtcgtgga ggagctcctg cgcggtgagg acgagcagtt ctccacgatc 41760
acggtggacc acacgccgaa gctgcacccg tccagctgga ccgcggtcgt cgacgccgcg 41820
ctgaccgcca gtggtgtgct ggtgatggac gagaactcga acgtcctgcg cacctgctcg 41880
cacctggagt ccctgtcctt cgtgggcccc ccgccgccca ggatccacgt acacaccgtg 41940
cgtgacccgc gcaccccgga caccatcagc atgacggtgg ccgacgagag cggcgccgtg 42000
gtctgcgagg cccgagggct gcgctacgtc aaggtccagg acatcggctc cggcgcggtc 42060
ggcccgcgcg acctggtgca cgaactcgcc tgggagccgg tggaggtccc cgcggacgcg 42120
ccggtcccgt cccaggccct ggtggtgggc ggcgccgccg gcggccccgc cctggtcgag 42180
gcgctcaccg cacgcggagt gcgggcacgg gcggtaccgg acgcgaccgc gatcggcgac 42240
gcctcgctca cctgcgccga cgtggtcgtc gtggcgcccg aggcgctgct gcccggggag 42300
gcacccgagc aggcggcgcg ccggtgcgcg cagctgctcg tcgacgccgt gcaacaggtg 42360
gcggccgtcc cggacgagcg gcggcggccc agggtgtggg cgctcacccg cgaggtacgg 42420
gcgggcgcga cggaggccgc gctcgcgcac gccccgctgt ggggcgccgg acgcatcgtc 42480
gccggcgagc gccccgacct gtggggcggc gtgatcgacg tcgcggagaa cgccgtaccc 42540
cagcaggtgg cctcgctcat cggagctctg ccgcacaccg aggacgtact gtcgctcgac 42600
tccgaaggcg tcaccgccgc ccggctgcgc caggtcgcgc ggcccgcgga gcgcgaaccg 42660
gtggactgcc gccccgacgg tacgtacctg gtcaccggcg gtctgggcgc gctcggcctg 42720
gaggccgcgc gccatctggt cgagcagggc gccaggcgtc tggtgctcat cggccggcgg 42780
ggtcttccct cccggtcgcg gtgggaccag gtggacgacc ccgccgtggc cgcgcagatc 42840
gccgaggtcg tcgccctgga ggcggcgggc gccacggtgc gtgtcctcag cctggacatc 42900
tccgacgccg aggcgacggc ccgtgccctg gacccgggcg ctctggacat gccgccggtc 42960
cggggcatcg tgcactgcgc cggtgtggtc tccgacgcgc tggtggagaa gaccggtgcg 43020
gcgaacctgg acaccaccat gggccccaag gccgacggcg cgatggtcct gcaccggctc 43080
ttcccggccg gcacgctgga cttcttcacg atgttctcct cgtgcggcca gctggcacgg 43140
ctcaccggcc aggtcagcta cgcctccgcg aactccttcc tggacgcgct cgccgccttg 43200
cggcgttcgc ggggggagac gggtacgacg agcttcgcgt gggcgcagtg gatcggacgc 43260
ggcatgggcg agaccaccgg cagggcgacg atcctggagg cggagtcccg cggactcggt 43320
ggcatcaccg tctccgaggc gctgcgcagc tgggcctacg cggaccggtt cgcgctgccg 43380
tacgcggcgg tgatgcgggt catgccggat cacacgctgc cggtgttctc ccacctgtcc 43440
gtcaccgacg cgggggcgca gagcgccgat gccgggggcg tcgactgggc caccgtgccg 43500
gccggggaac tgccggagct ggtgctgaag gtgacccacg agcaggtggc cgccgagctc 43560
aacctcgcgg tggacgacat cgcgatcgac cagcccctgc tggagctcgg tgtcgactcc 43620
gtgctcaccg tggcactccg ggtacggctg caccggtgtt tcgcggtgga cctgccgccg 43680
acgatcctgt ggagcaaccc gacggtccgc gccctcgcgg agttcctcgc cgccgaggtc 43740
ggcggtgcca ccgcggacgc cgaggagacc gacccggtcg ccggccttcc cgcacctcaa 43800
cagggatcgg gaactgccga gcagctcgac gccgtcgcgg ccgcggccgg gtgaccggcc 43860
ggcccggggt gacgacgatc tcgccacccc gggccgtccg ggcccgggac ggcgcgaccc 43920
acctgtcccg ggcccggctc gttgcagcct ttccagaggg tcgtcgcagt ataggtgcag 43980
tatttgtag tattgtttcgg caacgactgc gggttttcgt gtatgacgtc gcaccgcaga 44040
ggggaagccc cacatgccag gtcaggactc agtgctgatc gaggaatcga tacctggaag 44100
gttgcgcgcc gatcacttcg ctcagcacgg ttgggcacac ggccgcctga gcgtgcccac 44160
cgagggaagc gactgggagc aattcgcctc ggcctgggac gaactcgtcc cggaccccta 44220
cctcggcgag gagcggtgcc gtcgcaaccg gcggttcggc agggtgctcg cgcaccggga 44280
cggtgcgctg gagccgctgc gcggatccga cttcttccag agcaaggaga tcaaccgcgc 44340
cttcggcggc cggctgcgcg tgttcgagcc gctgaccgat gccgtactga cgggtccttg 44400
ccttccccgg ttgctgcgcg agaccgtcgc cgtcgtcaac gaggcggcgg ggaagcagga 44460
ctgggaactc ggcatccact tcatccgggt catcgcggat cctggtgaaa gctcggaacc 44520
ggcaccggag ggccgtcatt cggacggtca cgcctacgtg gccatacacc tgatcgaccg 44580
gcaccagtgt gccggaggac gtaaccaact gttccggaac aacgagtcgc gggcacagtt 44640
ctcggtcgtc atgaccgagc cgctggagac cctgatcgtg tccgacacga cgatggaaca 44700
cagcgtcagc gagatccggc ccgaagccgg caccgggtcc ggctggcggg acaccatgat 44760
cgtcgacttc aaccgtgtgc ccgtcccgga ggaagtgaca ggacgcacgt acggattcat 44820
ccggtgatgg gggattgatg actcacgtgg cagagacttc ggcacccacc cgaagcgaac 44880
cggatacgcg cgtgctcaca ctgcccggaa cggcatcggc accggagttc cgcctgatcg 44940
acatcgacgg cctgctcaac aaccgggcga cgaccgacgt ccgcgacctc ggctccgggc 45000
ggctgaacgc ctggggaaac tccttcccgg cagcggaact tcccgcgccc ggatccctga 45060
tcacggtggc cggcattccc ttcacctggg cgaacgcgca cgcgaggggg gacaacatcc 45120
ggtgcgaagg acaggtcgtc gacatcccgc cgggccagta cgactggatc tatctcctcg 45180
ccgcctcgga acggcgcagc gaggacacca tctgggccca ctacgacgac gggcacgccg 45240
atccgctccg ggtcggaatc tcggacttcc tggacggcac ccccgcgttc ggagagctca 45300
gtgccttccg cacctcccgg atgcactacc cgcaccacgt ccaggaaggg ctgcccacca 45360
cgatgtggct gacccgcgtg gggatgccgc gccacggggt cgcccggtcc ctgcggctgc 45420
cgcgttcggt cgcgatgcac gtcttcgcgc tcaccctgcg gaccgcggcc gccgtacgac 45480
tcgcggaagg ggcgacgaca tgaccaccac cgcaccgccc gtcgaactct ggacacgcga 45540
cctgggaagc tgcctgcacg gcaccctggc gacggcgctg atccgcgacg gccacgaccc 45600
ggtcaccgtc ctcggcgcgc cctgggagtt ccgccgccgc cccggggcct ggagctcgga 45660
ggagtacttc ttcttcgcgg aacccgattc gctcgccggg cggctcgccc tctaccaccc 45720
cttcgagtcg acctggcacc gcagtgacgg cgacggcgtg gacgacctgc gcgaggcgct 45780
cgcggcaggc gtgctgccca tcgccgccgt cgacaacttc catctgccct tccggcccgc 45840
cttccacgac gtacacgccg cgcacctcct cgtggtctac cggatcaccg agaccgaggt 45900
gtacgtctcc gacgcccagc cccctgcctt ccagggcgcg atcccgctcg ccgacttcct 45960
ggcgagctgg ggatcgctca acccgccgga cgacgcggac gtgttcttca gcgccagccc 46020
cagtggccgc cgctggctga ggacccgtat gacgggcccc gtcccggaac cggaccggca 46080
ctgggtcggc cgtgtcatcc gggagaacgt cgcccggtac cggcaggagc cccctgcgga 46140
cacccagacg ggactgcccg gcctgcgccg gtacctcgac gagctgtgcg cgctcactcc 46200
cggcacgaac gcggcgtccg aggccctgag cgagctgtac gtcatctcgt ggaacatcca 46260
ggcccagagc ggcttgcacg ccgagttcct gcgcgcccac tcggtgaaat ggcggatccc 46320
ggaactggcg gaagcagcgg ccggtgtcga cgcggtggca cacggctgga ccggggtacg 46380
catgaccggc gcccactcac gcgtctggca gcggcaccgg ccggcggaac tgcgcgggca 46440
cgccaccgca ctcgtccggc gcctcgaagc cgccctcgac ctgctcgaac tcgccgcgga 46500
cgcggtgtcc tgaccgggcc ggcacgcagc cggccgacga cacctccacc accgcacacc 46560
gaccgtgaag ggcaatcgtg gacagaactg ccatgcccat agccgtggtc gcctcccgga 46620
tacgccgcga ggagcggctg atcctcgaag cgttcgagcg gctcgggctg cgctgcgagc 46680
acgtcgatcc gcgcgggctc aggctgctgc tcgccgacgg ccggccgccc taccgggccg 46740
cgctcatgcg cgaagtgagc cacagccgtg ccgcctccgt agcgaccacg ctggaagcgc 46800
tcggggtccc cacctacaac aggccccgcg tactggacgt ctgcggcgac aaactgcgca 46860
ccgccctcgc cttccacagc gccgggctcc ggatgccgca ggccgcggcg acctggggca 46920
cgggggcggc cctggaagcg atgcccaagc tcgggtaccc cgtcgtggtc aaacccgtca 46980
ccggttcgtg gggacacctg accacgcgcg tccgcgacga ggagcagggc cgggccgttc 47040
tcgaacaccg tgccgccctg ccgaacccgc agcagcacgt cttcttcctg caggagcaca 47100
tcgacaaacc cgggcgcgac atcaaggcct acgtcgcggg tgaccgggtg atctgcgcga 47160
tctacaagaa cgcggtcgac gactggcgga ccaacaccgc catcggaggc gcggcgaccc 47220
cctgtccgat cacccccgaa ctgacggagc tcgccgtcgc ggcggcacac gcggtaggag 47280
gcgggttcct cggcgtcgac ctgctcatcg accagcagga gcgcctctac gccaacgagg 47340
tcaaccacac ccccgagttc cacggcgccg tcgacgcgac cgggatcgac gtcgcgcaag 47400
gcatcgtcga gtgggccgtc gcggagatcg aggcacagga aagggcggcg gcatgagcgc 47460
gacggccggt cccccctcgt tgctcgcgct gtggagcgcg ccccgctccc gctccacggt 47520
gttcttccgg atgatggtgg aacgccagga cgtcctggcg ctgcacgagc cgttctgcaa 47580
catcgccaac gacggcagca ccgtgatcga cggccgcgtg gtgcacagca cggacaccct 47640
gacggacacg ctcatcggac tcgcggcaca gcgaacggtg ttcttcaagg acaccaccga 47700
ctgcttctac gactccgtgt tcgcacgtcc ggacttcctc aagagcgggc accacgcctt 47760
cctgctgagg aacccgaagg agatcatccc gtcgtacgcg gccatcaagc cggacatggc 47820
ctgtcacgaa gtgggcatcg cctacctcca ccggatccac caggcggtac tcgacgccgg 47880
cggcagtgcg gtgatcctgg actccgacga cttcgtggac cacccggagg cgaccgtccg 47940
gcagtactgc gcgacggtgg gcctgccctt ccgcgccgac gccctccaat ggcagccggg 48000
tcagcggccg gagtggagcc agtccgcgcg ctggcacacg gatgtcagcg ccacgtcgac 48060
gatacggcgc cgcgagcgga cctacgacag gacgacggac aacgacccgc tgctgcaacg 48120
cttctacgaa caccacattc ccttctacga gtacctgtgg gagcggcggc tccggatctc 48180
ctgacgggac ccgggacacc gactcgccga tgaaatgggc gctctggccg atcagatccg 48240
agaggtacgg cagatgatta tttccaagcc cccgttctgc ccggtgcagt tccaggactc 48300
cgacctgctc aactccggat tccacgccgc cacggacatg cacgcgctct gggccgacct 48360
gcgtgagaac cagccgctct accgggccga ccccggcggt ggccgggagc cgttctgggt 48420
cgtgacccgc cacgaggacg tcagccgggt gctgcgcagc cacggggagt tctcctcccg 48480
gcggggcacg atcctgtgcg tgctcgacct caacaccgcg gacatcgcgt ccgacaacat 48540
gatggccgac accgatccgc cccggcaccg ggagttcagg gagccgctca acaaggcgtt 48600
cgccccctcc gtggtcgcca ctcaggagcg gctgctcaga gaactgtcgc aggacctgat 48660
ccagtcggtc ctggaagccg gcgtgtacga catcgcccac aagacgatga tgttcccgat 48720
ggccgtcacg ggcacgctca tgggcctgcc ccccgagtcc tgggaaaggc tcgcggagct 48780
ggtcatgatg acgatcgcct acgacgaccc cgactactcc gccggcagtg cgcaggccac 48840
cgtgcgccag gcccggcacg agctgttcga gtacttccag aaggagtacg cgcagcggag 48900
ccgggacgac gtcgaccccg acgtcatcgg cgcgatggtc ggcatggatc tctcggaagg 48960
accgatgacg caggagcagg tcctcctcaa cgccttcgtg ctgctcatcg gcgcgaacgt 49020
caccacgccg cacgccctgt gcaccctgat gtccgtgatg gccgaacacc cggagcagta 49080
ccgggccgta caggagaacc ccgagctgcg ggcgtcctgc ctccaggagc tcctgcgctg 49140
gtcctccccg gtcaccgcgc tgatgcggta cgccgtcaag gacgtggaga tgcacggccg 49200
aaccatccgg gcgggcgagc cggtcaccgc gtggatcggc gcggccaacc gggacgagag 49260
ggtcttcccc gacccctaca ccttcgatgt cgcacgacgg ccgaacaagc acctcgcctt 49320
cggcctggga ccgcactact gcatcggcgc caacctggcc aaggtcggcc tcgacatctt 49380
cctcgacgaa ctcctggagc gggtggagag catcgagatc gccggcgagg tgaagcacgt 49440
cgcctcccac ttcgtgcccg gctacaagag catgccggtg cggttcactc ctcgtgaggc 49500
acacgtacgg tgagcagcta ccccgtgcac cgctccgacg cccggctgtg gttcgacggc 49560
cggttccgct ccgccacggc gacacacagt ctgtactgcc tgcccttcgc cggtggttcc 49620
gcgacctact acgccgactg ggcgccccac tgcgcgagcc ccgtcgaact ggtgccggtg 49680
cagctgcccg gccgcggtgg ccgcatgacc gagtcgtcgg ccaaggatct ggtgcagctc 49740
gccgaggaga tcgccgacac catcgccgcc gaacccacgc ggaccctcct gtacgggcac 49800
agtatgggag ccatgctggc gttcgaggtg agccgacggc ttcagacgct gaaccggccg 49860
gtgcggcacc tcttcgtcag cggccgcccc gcgccgacga tcgtgcgtcc gatcgccccc 49920
gtcagccacc tgccccgcgc cgagttcatc cagatgctgc gcgactacgg cgccgccgac 49980
cagacggtct tcgagcacga cgaactgctg gacctgctca tgccgatgat ccgggcggac 50040
ttcgccatga tcgagaacta ccggtaccag gaggcaccgc ggctctcgtg cccgatctcc 50100
gcctggtgcg gagacgccga cccggaggtg cctcccacgg cgatgcgtgg ctggggcgac 50160
cagacatcgg gagagttcac cctctccgtg ctgcgaggcg ggcacttctt cctgaccgag 50220
caccgggccg agatcatgcg ggccgtactc gccgcggtcc gtcgagcgcg ttgactaccg 50280
acaaggagtg agagcgtgga cttcgatgtg ccacctcaga gcgtcgactt cgcgaacgag 50340
gtcgaggaga ttctgacggc cccggagacg gaacggctgc tgcgggagct cgccacccgg 50400
caggacggca tggacggcga cccccgtgcc ttgtaccggc atctgggcga cgcagggatc 50460
ctcgctccct cctggccggt cgaatacggc ggcaggggcg ccgacttcac cgcgaccgtg 50520
gcgctgctgg aaaagctcgt cgcccaccgg ataccgcaga atctgtactg catctcggtg 50580
cagaacgtcg gctccctgat cctggccgcg ggctcggagc agcagcgccg gacgctgctg 50640
cccgccctgg cgagcaccga actcacgtcg tgcatcctgt tcaccgagcc cggcaacgga 50700
tccgacctgg caggcatcac cacgaccgcg gtgcgcgacg gcaccggctg ggtcctcaac 50760
gggcgcaaga cctacaacct caagagcgcc tacgccgaca tcgcgctcat cgcggtacgg 50820
accgatccgc agagcagcca gtacgaggga ctcaccctct tcctcgtccc gctggacgca 50880
cccggtgtcg tcatccgccc cattcccagc ctggccgacg agcagttcca cgacatctgg 50940
ttcaccgatg tgcgtgtcga cgagtcagcg ctgttcggcc aggtcggcga gggctggtcg 51000
ctgatcactc agatgttcgc cgcggagcgc acggggctgg actactacgc gcggggacgc 51060
cactggctgg atctcgtcgc gcagcgtgtc gacaggaacg gtgcacggca gtccgatgcc 51120
gtacgcaccg aactggcccg tcacgaggcg cggatcaccg cgagcaaact gctgagctgc 51180
caggtcatgc agaaccttca ggacggcagt cccgacatcg ccgaatcctc cttcgcgaaa 51240
tggcactgca gcgaatcggc ccagcgcatc gcctggtggg ccctggacac cctgggccag 51300
gacctgctgg aacccgggcc cgaccccggt gaccgcactc tggaggcagc cttccgggag 51360
gcaccgggaa tgaccatctc cggcggggcc tcggaggtca tgctggacat tctgagcagc 51420
gcccgggtat tcacccacgc cccgagggag gactgaccgc gatggacctg cgacctgatc 51480
cgttccagca gcgtctgcgc agcgccgtgg acacggtcct cacgcgcacc ggcgacctcc 51540
ccgctcaact ggcggacatc ggcgtgccct ccctcggcgc accggagcgc ctcggcggct 51600
tcgcgctcgg actcagcgcc gacatcgtcg tgaacgaacg cctcggacac ggactcgaac 51660
cgctcgacag ctatcgggcg accgcgttcg cgctcgacct gctcgactgc gacggcgcac 51720
ccacggaact gctggccgag atcctcgctg acctgtacaa gggcacccgg cacgcggtgg 51780
tcgtgggggc gcgaggcggc ggcgccgtac aagtgcggga cggaggccgg ctgtggggcg 51840
agtcggaatc gctgcctgcg ggtgacatcg cgctgtgcat cgtgcgaacc gccggcgagg 51900
acggcgccgc atggcgtctg gcgattccgg aacccggcac ctgcgtcacg gagccggcgg 51960
agcatctcgg catacccgcc tcgcgattga gattccacgg tgcgccgtcc cagtctctgc 52020
cggtgaccac cgctcactgg cagcgggccc tggcggcggc gcggatccgg caggcggcac 52080
tgctgctggg catcgccgac tggatcaccg atgtggcccg cgcccatgtc aacacccgca 52140
cgcagtccgg caaaccgctc gtggagcggc agacggtcgc ccaccggctg gccgtcctga 52200
tcggcgaggg ggagggctgg aagctcctgc tgaacaaggc ggcctgggag ttcgaccgcg 52260
gcgggcgccc cgacgcggcg gcgatgctgg cggtcgccgg cgaacacgcg cagctcgcca 52320
gccgcatcgc cctccaactg cacggcgtac ggggaatgct cgcgcactcg acagcggcat 52380
ccgtgtaccg gatggtggcc gtggagtcgg cgcggctggg cacccccgcc gcgctgtggc 52440
gggaagccgg atccgccgcc tgactccggt cggcccacat tcaccgttcc gctggcatcc 52500
cgggtttctc ggtgtaggtt gacatgcaga ttctcatgca gtgtgtggac tgtgggccgg 52560
aaggaacggt gtgaaggcca gctggagaca ggtatttgcg tggcggatgc agcggcagtt 52620
cctggagccg cgtacccagc cgagtgcgag tgacgtcgtc ggcagactct gtggcgtgca 52680
ggcacaggtc tggagtgtcg ccgagctgaa cgtcgcactt cggcaggcgg cacccgaccg 52740
ggaaagcgtc aaccgggagg tcgccgacct ctcgttgatg aagacctggg cgatgcgcgg 52800
aaccctgcac ctcctgcggc cgagcgaggc cgggccctac ctgtcgctca tggcgaacac 52860
gggaagctgg ctcaagccgt cctggacccg ggcctccggc gtcacgcccc gacaggtcga 52920
cgagctgacc gaagaggtgg cgggaatcct cgacggagtg gtgctcaccc gggatgagct 52980
cgtcacccgg ctcgtcgccg acaagcggtt cgtgagcatg gaggagcgac tgcggtcggg 53040
gtggggctcc gtgctcaaac cgctggcctg gcgaggcgtc ctgtgccacg gcccgaaccg 53100
gggcaacaag atcacgttca cgctgccggc gagccagttc ggtgcggact ggggcaagat 53160
gcccgaaccc gacgaggcgg cgcccacggt gatcaaggcc tacctgggcg cgtacggccc 53220
tgcgacgatc gagaccttcg accgctggct gtcgctcaac agcacgagca agccgaagct 53280
gcgcaagtgg ttcggcgaca tgggcgacga gctcaccgag gtcgacgtcg aggggcgcaa 53340
ggccttcgtc ctgacggagc acgcggagga actcgccgcc accgccccgt gcaccggcat 53400
caggctgctc ggcggcttcg accagtacct cctcggcccc ggaaccaagg acgaggtcgt 53460
cctcgcaccg gaacaccgct ccgcggtgag ccgcgccgcc ggctggatct cccccgtggt 53520
cgtgaaggac ggcagagtcg tcggggtgtg ggagatcgtc gaccaggagc tggtggtgac 53580
cccgttcccg gacaccgaac ggctccctgt gaaggcggtc gagaaggagg ccgcgcacgt 53640
cgcccgcgcc agcggagtga gccggctgcc ggtgcgcatc gtctgacgac ctggcccgac 53700
cgctctgccg cgctcgcccg cagctcacgc gaagcgggcg cggtagagat cgcggagcag 53760
ggcgatctcc cctccgtgat gcatcacttc acgattgacg tgcaggagca gggtcgccat 53820
cggccgctct ccccacggcc cctcgacatc tcctatgggc cgcgccagtt ccgcgtcgtc 53880
gagggagagc acccccttgc gccatgcgtg gtagtgctcg tgcagcgccc cgagcccgtc 53940
cgcggccgtg acggggatgg acgcggtctc caaggtcagt gatccgtcgc cgaagtgagc 54000
gctggaccgg atgcccagta cgtgaaaggc gacgtggcac atgcgccagg cgatcgtcgt 54060
cacgggaggc ggctcggggg cggggcgtcg caggtcgagc gaggcccttc cgtcctcgcc 54120
gacgcggacc gaccagcagt ccgccaccgg ctcccacaga tactcgtcgt cggtgagacc 54180
ggccagcctg ggcatcaagt ggtggtccca gtacaggtcg agctgatcga ggacctcggt 54240
tcgccagatc atccagtact cctgccgaaa tcgtgatgtc ggtgatgaac gcatacgagg 54300
gtaaccggtt cagcacagtc tgcactcaca tcaccaccga cagtagtcgt atggggtatt 54360
gatcgtcgtc gaagacgccg cgcgactcgt agattccttg tcgccactgt tgtcttggtg 54420
taaaaagacc gcatgatggc gtggctctcg gaggacgtgg ggcgggggag gacgcacgta 54480
ccactccaca tgacgggtca cggaaatcat ttgcccacaa cgcgatctgt aatgtccgta 54540
ccggcgtgcc tggtaccgag ggagcacccc gatgagtgat cgcgtccagg tgaccgtcga 54600
cgcgcacgac ccgcgcgcgc tgtcgtcctt ctggagtgcc gcattgggct acgtccacgt 54660
cggcgcggag ggcgagcgcg acacgcgcgc ggtgctggag gaccccgagg gccgggggta 54720
ccggctgtcg ttccggcggg tgccggaggc cgagggcacc gggaaccgcc tcagcatcgc 54780
ggtccacccg gcccccgggc tgacgggcga ggagcggatg acggcgctgg aggccgagtg 54840
cgagcggctc ctcgcactcg gggcgacgcg cgtacgccgc caagagcctg cctccgtgat 54900
cgacaccggc cacatagtga tgacggacct cgaaggcaac gagttccgtc ttgagtgacc 54960
ttgcccaggg gcctgcgcag gcggtggcgg ttgcagaagg tctcgatgaa gttgaagccc 55020
ttcgggcggc cgggggcccg gtcggatgat gtgcgggtgc cggtcccttc tttgagcagg 55080
gtccagaagc tctccgcggg ggagttgtcg aagcacgacc cggtgcgtcc gcagctctgc 55140
cgcggtccga acgtgcctgt gcggtcccgg aatcgggccg acgtggattc acctccgcga 55200
tcgctgtgga tcaggcagcc gtcctccggt ccactccggt cgtgggtcat gccgagtgcg 55260
tccacgacga gttcggcgcg gtgatggtgg gccatcgcgt agccgacgac ctcgcggacc 55320
acggtccggt gccgcgccgg ctgcgggaac gtcggacgct tgccggaccc ggaacggcct 55380
cgtggtgcac ccacccggga tgacaggacc gccaccttct acgaggcagc ggtcccactc 55440
acgtccttcc cgctcagggc aagatccgat cggagacgga acctagtacg cgagctgccc 55500
gatgcctccg tcgacgcgca gcacggtccc cgcggtgtag ccggattcgt ccgaggcgag 55560
gtagaccgcc gccttcgcca gctcggtggc ggtgcccagg cgatgcagag gcacggtctg 55620
ccggagttcc tcgtagaggg cggcctggcg ctcggggccg agggacgcga acgcattggt 55680
gatcgtcgga cccgggctca gtccgttgac gcggatgccc cggtccttca gctcatgggt 55740
cagccctcgc gtgtaggaga gcagacccgc cttggccgcg ccgtagacgg ccgcgttgtc 55800
gtgtccgatg tacgcggaga ccgagcccac gaggatgacc gaggactgcc gcgagaagag 55860
cggcagcagg tccctgatca ggaagaacgg cgacttcaga ttggtcgcta cgagcctgtc 55920
gaacgcctcc tcggtccatg cctcgatggg gaggtgggtg atgtcggcgg cgttgctcat 55980
caggatgtcg agcttcggcc actcctcccg cagacgcgcc gcgagcgccg cctggccggg 56040
tacgtcgccg gcgtcactga cgatggtcag cagcgggccg cccagctccc gggccgcctc 56100
ctccagtttc tcccgcgaac ggcccgtgat ggccacggtc gctccctcgg cgaggaattc 56160
gcgggccgtt tccaggccga tgccgctcgt ccccccggtg atcagtgcgt gctttcctgc 56220
aagtcggtcc attgtcctgt ccctcttcag atcgcggtgc ggccaccgtc ggcggccacg 56280
accgcgccgg tcatgtaact ggcccggtca ctggcgagga aggcgatcac gtgggccacc 56340
tcggccgggt cggaggtgcg cttgagtgcg gtggtcagac ccatgccgcc catgtccggg 56400
cccattgccg cgaccacctt cgaggtgcgc atcgggcccg gggcgacggc gttgacccgc 56460
acgttcgagc cggcgaattc cgccgcccag gtgcgggtga gcgattcgag cgcggccttg 56520
gtggcaccgt agacggccat gcccggcatg ccgaggcccg ccgcggtgga actgacgttg 56580
acgatgctgc cgccgccgtt cgcggccatt ttctccgcga gcagggcggt gagcaggaag 56640
ggcgcccgca cattgacggc gaaggcggcc tcgtagctcg cgaggtcctg tccggtggtc 56700
ggggagaacg tcatcacgct cgcgttgttg accaggatgt cgacggggcc ggcgtcctcg 56760
gccagctgcc gcacgctcgc cgggtcggac agatcggcgg cgatgaaacg caccgccccg 56820
gcggagccgc cgccggctgc gcggaggtct tccacgacct gggcgccgcg ctgcggatcg 56880
gtgccggtga ggatcaggcc ggcgccgccc gcggccagca ccccggccgt gacgtgcccg 56940
agtccgccga tcgcaccgga tccggtgacg agtgccgtct tgcctgcgaa ttccatgctt 57000
cgcccttcat gccggccggc gagcgaccga cttctgtgca gtcgttcaag aagtgtcgcg 57060
acggaagcta cgacttcttg aacgccaagt caagaagtcg taggatctgt cccgtggcac 57120
gaaccggacg cccccgcgag ttcgacaagg aacggacgct ggagcgcgca ctcgaactct 57180
tctggtcccg gggatacggg gcgacgtcga tccaggacct ggtcgacgcg ctggcggtcg 57240
aacgcggcag cctctacggc gcgttcggcg acaagcgccg cttctacctc gaggccgtca 57300
ggctctactg ggaggtgtac gagcggcacc tgaccgccgc gctcgacacc cgccccctcc 57360
tgcccgccct gcgggagatc ctgacccatc ccgcgcggct ggacgaactg atctccgacc 57420
tgggcgtccc acagggctgc ctgatcggca acacgaccgc cgagctcgtc ccccaggaca 57480
gcgaagccac gcagatcgtc acccgctcgt accaccggtt caccgacatc gtcgccgctg 57540
cgctccgccg cgcacaggcc gacggagagg tcacggacac ctgctccccc gaggcccagg 57600
cacaactcct cctctacatc gtccagggcc tctcgctcgt atcgagggcg gggctcgaca 57660
gggcggcggc cctggcggcg atcgacacgg cgctggacgg actgcgggca tgaccggtcc 57720
cccgggcatg cgaagagtcc ctcgctgtgc aggcgtaccg agatccgcgc tgccgtcagc 57780
cgcacacacg tctcccggac agtgcgacac gcggcggccg tgatcgtcgg tggatcgtgt 57840
gtcagacctg tctgggaggc tccgggcatg gagatgacag tgcagctgac gatcgactgc 57900
tccgatccgc ggacaatggt ggcgttctgg gcggaggccc tgggctacgt gcccgagccc 57960
gcgccggacg ggtacgcgac gtggggcgct tactgggagg cgacgggggt gcccgcggaa 58020
gagttgccgg ccggggcagg cgacaccgcg gagtcgatcg tcgatccggc gggtcgcggg 58080
ccccgtgtgt ggttccagca ggttccggag ccgaaggccg ccaagaaccg gtggcacttc 58140
gacctgaagg ccggtggcgg ccgtgacgtc ccgttggacg tccgcgcgcg gcgggtcagg 58200
gccgctgtgg accggctggt cgaagccggt gccaccgtgg tgaagatcaa ggatgagccg 58260
cgcacggggc tttacgccgc ggccatgcag gaccccgagg gcaacgagtt cgacgtcgtc 58320
tgagggctgt tgtgatggcg ccgaccacaa tcactcgtcc acggtcgtgg ccggtactgc 58380
catcgagcat gaccttggca tgccatcggg ttccccgcag tgactctgtc cgcctgtgcc 58440
ctcggccacg gcgggtccct cctgggcggc cggaccgccg gtccgtccgg ccgacgacgg 58500
tgccttcctg gcctcggtcg ccgtcttcgg catggtgagc gtgtgggagc gctcgctccg 58560
accggtggcc tgctgatcga ggcgcggctg gtgaaaggca cgccaacgcc ctggccgacg 58620
ccctcccctc cgcccaggca ctggccctcc tcggcaccct gcacgccagc gggaagcgca 58680
tcgactcctc ctgggtgcgg gcggcccggc gtggcccgaa agcgggacca gtccggtcga 58740
ccgtgcacgg ccgggcatag gtagcgtggt ccccccgcag cgttcagccc atcaggagtg 58800
gcacccacgc acagggttgc cggccgggac gcttacctcg tgtgaggtct cggtcacggc 58860
gaccgcttcg gagccttcgc ggcgcatccg ctcgaccccc caggcgccga gcggtccgag 58920
cgcttggttg agcgtgcgcc cgtgctcggt cagggagtac tccacccgcg gcggcacctc 58980
ggcatagacc tctcggtgca cgagaccgtc ttcctccatc tcgcgcaggt gctgcgtcag 59040
catcttctcg ctcacccccg gcagaccgcg acggagctca gcgaaacggc gtacgcgatg 59100
ggcgtcgagt tcccagagga tcagtccctt ccacttgccg ctcaccacgt cgagcgcagc 59160
gtcgatgccg cagatgtagg gcccgcgtct cggtgccttg gccatcactg gtccccctta 59220
cggaaaggta agtaccgcag aaaatagtgg gtacttctga gagtagtgac actctccgag 59280
catggagggg tgaacgaaca acagaaccac accaccaggc aggacagcac tgtcaccgtg 59340
atcgggctcg gcccgatggg ccaggcgatg acccgcaccc tcctcaccgc cggccacccg 59400
gtcaccgtct ggaaccgcac cgccggtcgg gccgacggcg tcgtcaccga cggagcgacg 59460
ctcgcggcga cacccagcga agcggtcgaa gcgagtgacc tcgtgatcct cagtctcacc 59520
gactaccagg cgatgtacga catcctcggc agcgccaccg catcgctcgc cggccggaca 59580
ctggtcaacc tgagctctga cactcccgac cgttcacgcg aggcagcaac ctgggcagcg 59640
ggccacgacg ccaccttcct caccggaggt gtcatggtcc ccgcgccgat ggtcggcacg 59700
gaggcagccc atgtctacta cagcggccgc gaccaagtga tggagcgcca cctggcggcg 59760
ttgacactgc tgggaacacc aaagtatctg ggcgaagacc cgggcctcgc ccagatgatg 59820
tatcaagccc agctcgcggt gttcctcacc accttgtccg gactcatgca cgccaccgcg 59880
atgctgggca ccgcaggaat gaaggccaag gaagcgctgc cggagttgct ctcctccgcc 59940
gactcgatcg gcgacatcct gagggctggt gaagagaacc ccggcaccgc gcttgatgcc 60000
ggagaacatc ccggcgacct cagcacggtc acgatgatgg gcgcgacatc cgaccacatc 60060
gtcgagacca gtacgtcact cggcctcgac ctcgcgctcc ccctggccgt gcaggcccac 60120
taccggcggg cgatcgagaa cggacacggc agcgacaact ggacccgcat catcgacagc 60180
atccgaggac cgcgctgacc cgtgcgcggc cacccgacgt tccggccccg gccggtgtgg 60240
ccggggcgat acgtcagcgc caccgtcgcc gttgcccata cggtcccctc ccgtgggaag 60300
cggtgccggt cttcgtcgtt gagccggcgg agtcgtacac gggctcgacg cgcccgcgtg 60360
aaagcccggt cgtggcgtcg gtgacgccgc tgtggagtac ggcaagcctc tgctgctcga 60420
ggtcactcct gctgcacccg aatgccgtcc ggatcctccg gatttccggt ccgcgccccg 60480
gggccggcga gagacgccgc cggggcggtg cctctcgccg gagaccgtca gccgggcaac 60540
cgggcgaacc ggagacctgg acagccgggc atgccgccaa cctcagctcg aagcaagctg 60600
gttggcgatc tccttgtgcc acttctcggc gtcggcgggg tcgttcaggg atgccgccca 60660
cgcctcctcc gtcatgtgca ggttcccggg gcccgacttc agcgcgacga gcagttcccg 60720
ggccgaggtg cgcagttcgg gcgaggtgcc cttgccggcc gggacatcgg ggcccgcggc 60780
gaggccgtag agggtgcgcg ccatggcgtc gcgcttgctg tgttcggcct tcttccagaa 60840
cttcagcgct tccttgaggg cgtcctggcg ggaggcgggc tcggctgtcg tctgctcgcc 60900
ctcctccgtg ggcggggccc agcgctcggg gtggcgggcg ccgtcgtaac gcttcttgcc 60960
ggcgcgcccg gccttgtacg cggtgaccga gagcaggagc cccgcagctg tcgcggccag 61020
cgcccagccg gccgggttgc tcgccagcgt ccccgtggca gccgctgcga tcgccaccgc 61080
gccgccggcc gacctcacgc cgtccccgac cgcgccgacc ccctgcttcc agagcttgtt 61140
gcgctgcttc gccagcgcgt agtcccgggc ggtgttgagg gcgacggtgt cctgtgcctt 61200
cttgacgtcg tcggcggcct tctcgatgct cgtgaaggcc gcaccggtct ggtccagggc 61260
ggcgcccagc cggtccgcga ggccctcgtc gtcgccgtca tgggcacgct ccagcgcgac 61320
atacgcctgc gcgagcgccc agtggctgtc cacctccgcc ttgtggagcc ggttgatctc 61380
gggctcctgc accggcgcgg gcacgtcgat cttcttgacg ttctggtact tgcgcgtcgt 61440
cagcccgaac cggcgcgccg cacggacgct cttgacgccc gcgacggcgc cgaccatccc 61500
gctcccggcc tcggcgacgc tcgccgccgt cacgaggccc tccgcgtgca gagcgttcct 61560
ggcgacgccg atgaagtcgg aggcgtcggt cgccgtgttc tggatcgcgt cgagcatctt 61620
ggagggatac ttcttgcgcg gggcatggga ggagggaccc gtcgggttct gcttggactc 61680
cttgtacgcc ttggtggcgt cgatgacgtt gttgaccgcc gtgaccccgt cggtgagccc 61740
catctcggca ctggccgctg cggcggacgc gccgacctgc tccgaaaagt gcgtgtcggc 61800
gttctgggac gacttccagt tcatctccgc gttggccggg accatgccgc gcaggacgaa 61860
ggtgtcgagg acgtcgaggt gcttgttggc ccgctccagc gcgccggcga tccgctcccg 61920
gatgttcagc ccggtacggg acttcttctt cgcttccggc ttggcctgct cgtcggcggg 61980
cttcgccgtc tcgtcgttcc ccgcgagacg ctggacgacg gcggacgcgg ccttgttgcc 62040
ggcgagcgcc tgtatggacc ccatcatcgg tccgacgctg tggcggtccg gcgcccccgc 62100
cagatgctcc gggccgcgta tgagaggccg gggctgtgga acggccggct gcttcgtacg 62160
gttgtcgaaa gcagtgttac tacccaccgg tgtcctttcc gtgctcgtgc tgagaccggg 62220
tgcgtggcaa gtcggggcat ccggcggggc cttgcttccc cacagtatcg actgccccat 62280
gactctcatg gaaggtgttt ctcgtctcgc gttcattctg tgtatcggtt ctgtacaccg 62340
gttcgtgacc atcgcattgt tctggccgct ccgggccccg gcatcagcgc cgccatacca 62400
cagcagccac cctcgccgga cgaccccgcc cccgcgggtc cggttgcgga gttcttgctg 62460
catatcgacg gtgagcgggc ctggttccgt tggaacggaa ggggagccgg tgctcgcggt 62520
tgtactgcgg cgcccggtgc cggacccggt cctggaggat gcagcgggcc cgctgggctg 62580
ccatcgacgc cgggggcgag gcgtacaccg ctctgctgaa ggggcgcccg ccccctggcc 62640
cggttccgct gcccgcagtg cgaagccgtc tggtacgtca gtgacacccc gatggggaac 62700
cggaaacggc cggacagccg caactgctcg ccgcgctgcc gtacccggcc gcgcgccggc 62760
ggcgactgcg aatacacccg cgctgaccgg gccccccatg ccatgaccct gcgcacgacg 62820
gccaccacct tcgactccag cgcggcagag cctggtgaag agcacctctc cgaaggagcc 62880
gcattcccca ccagatggcg gtcagcaccg ttgtcgactg ggcatacggc agggtcgcgt 62940
actacacggt cgcaacgccc aggaagcgca gcaccgccag gacgcggcgg tggtcggcgt 63000
cggccctggg caggtcgagc ttggtgaaaa tgctgttgat gtgcttggcg acagcgctct 63060
cgctcaccac cagctcggcg gctatgccgg agttggaccg gccgcccgcc atcagctcca 63120
gcacctcccg ctcgcgcggt gtgagccggt cgagcggatc gctgtgccgg cgcaccagca 63180
gctgcgcgac gacctgcggg tcgagcgcgg tgccgccggc cgccacccgg cgtaccgcct 63240
cggcgaactc ctccacgtcg gcgacgcgtt gcttgagcag atagccgacg cccgaggtgt 63300
tggcggcgag cagatcggcg gcgtaccgct cctccacgta ctgcgacagc aggagcacgg 63360
cggtacgcgg gtactggcgg cggatcacca gcgcggcgcg tactccctcg tcggtgaagc 63420
cgggcggcat gcgcacgtcg accacggcga tgtcgggccg gtgctcctcg acggccgcca 63480
gcagcccttc cgcgtcggcg acctgcgcgc acatctcgaa gccggccgcc tccagcacct 63540
tgaccacgcg 63549
<210>2
<211>1263
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val Thr Thr Ala Thr Ser Ala Ala Val Pro Gln Leu Ser Leu Glu Thr
1 5 10 15
Val Ala Glu Ile Ala Ala Gly Thr Ala Gly Val Arg Asp Ala Ala Ala
20 25 30
Val Arg Asp Arg Arg Leu Arg Val Thr Ala Thr Arg Arg Ala Ala Ser
35 40 45
Gly Pro Gly Ser Gly Gly Gly Glu Ala Gly Arg Gly Thr Ala Val Pro
50 55 60
Ala Asp Arg Pro Ala Leu Val Arg Gly Pro Glu Pro Val Leu Pro Pro
65 70 75 80
Asp Ala Val Pro Thr Leu Pro Gln Ala Leu Ala Arg Ala Ala Arg Val
85 90 95
Ala Pro Gly Arg Gly Thr Thr Tyr Leu Leu Pro Asp Gly Ser Thr Asp
100 105 110
Arg Gln Thr Tyr Ala Glu Leu Tyr Glu Asp Ala Arg Arg Met Leu Gly
115 120 125
Gly Leu Arg Arg His Gly Leu Ala Pro Gly Asp Pro Val Leu Leu Gln
130 135 140
Cys Ala Asp Ser Arg Thr Phe Val Thr Ala Trp Trp Ala Cys Val Leu
145 150 155 160
Gly Gly Phe Leu Pro Thr Pro Leu Ala Pro Ala Pro Glu Tyr Ala Thr
165 170 175
Asp Asn Ala Ala Val Arg Lys Leu Ala Ala Ala Trp Asp Leu Leu Gly
180 185 190
Arg Pro Pro Val Ile Ala Asp Pro Gly Leu Cys Glu Gly Val Arg Ser
195 200 205
Leu Ala Gly Arg Leu Pro Gly Gly Glu Ala Pro Arg Val Leu Ala Ser
210 215 220
Asp Ala Leu Pro Asp Pro Glu Pro Ala Asp Trp Thr Val Pro Asp Pro
225 230 235 240
Asp Ala Leu Ile Val Asn Leu Leu Thr Ser Gly Ser Thr Gly Thr Pro
245 250 255
Lys Cys Val Gln His Arg His Arg Thr Ile Val Ala Arg Ser Tyr Ala
260 265 270
Ala Ile Ala Ala Asn Gly Phe Thr Glu His Asp Val Ser Leu Asn Trp
275 280 285
Met Pro Leu Asp His Val Gly Gly Met Ile Met Tyr Asn Leu Arg Asp
290 295 300
Val Phe Leu Ala Cys Glu His Val Asn Val Arg Thr Glu Ser Val Ile
305 310 315 320
Arg Arg Pro Leu Leu Trp Leu Asp Cys Leu Asp Arg Phe Arg Ala Thr
325 330 335
Asn Thr Trp Ala Pro Asn Phe Ala Phe Ser Leu Val Cys Gln Arg Ala
340 345 350
Glu Glu Ile Ala Ala Gly Ser Trp Asp Leu Ser Arg Leu Thr Asn Ile
355 360 365
Cys Asn Ala Gly Glu Ala Val Val Ala Arg Thr Ala Leu Arg Phe Val
370 375 380
Glu Leu Leu Ala Pro His Ala Leu Pro Ala Asp Ala Met Val Pro Cys
385 390 395 400
Trp Gly Met Ser Glu Thr Ser Ser Gly Val Thr Tyr Thr Arg Met His
405 410 415
Arg Asp Asp Ala Ala Val Gly Thr Val Ser Leu Val Pro Ala Ser Leu
420 425 430
Asp Gly Glu Leu Arg Glu Leu Pro Ala Gly Thr Pro Gly Ala Val Val
435 440 445
Val Thr Glu Val Gly Ser Pro Val Pro Gly Val Ala Leu Arg Ile Val
450 455 460
Asp Gln Glu Gly Glu Val Leu Pro Glu Gly Arg Val Gly Arg Leu His
465 470 475 480
Val Ser Gly Asp Thr Val Leu Ser Gly Tyr Ala Tyr Asn Glu Arg Ala
485 490 495
Asn Arg Glu Ser Phe Thr Ala Asp Gly Trp Phe Asp Thr Gly Asp Leu
500 505 510
Gly Phe Leu Arg Asp Gly Arg Leu Phe Leu Thr Gly Arg Arg Lys Asn
515 520 525
Met Val Ile Val Asn Gly Ala Asn Phe Pro Ala His Glu Ile Glu Ala
530 535 540
Val Val Glu Gln Val Pro Gly Val Val Pro Ala Cys Ser Ala Val Ala
545 550 555 560
Gly Val Pro Asp Glu Asp Thr Gly Thr Asp Ala Leu Val Val Phe Phe
565 570 575
Val Pro Gly Thr Asp Asp Val Pro Ala Leu Val Asp Ala Ile Arg Ser
580 585 590
Ala Leu Ala Arg Asp Leu Ala Leu Arg Pro Thr Tyr Val Val Pro Val
595 600 605
Thr Ser Arg Glu Phe Pro Arg His Asn Gly Gly Lys Val Gln Arg Glu
610 615 620
Arg Leu Leu Glu Gly Trp Arg Glu Gly Leu Phe Asp Asp Arg Cys Tyr
625 630 635 640
Gly Gly Gly Thr Asp Gly Val Ala Pro Asp Glu Ala Pro Asp Ser Glu
645 650 655
Ala Ser Val Pro Ala Leu Ala Val Val Trp Glu Pro Ala Asp Ala Arg
660 665 670
Leu Arg Gln Arg Pro Asp Gly Pro Leu Val Ala Cys Val Ser Gly Asp
675 680 685
Val Pro Asp Trp Leu Pro Val Ser Thr Thr Ala Gln Ile Val Val Gly
690 695 700
Pro Asp Ala Val Glu Leu Ala Ser Ala Leu Glu Arg Ala Phe Ala Thr
705 710 715 720
Ala Gly Glu Gly Pro Thr Asp Glu Gly Ala Val Pro Gln Val Leu Phe
725 730 735
Val Ala Ala Ser Asp Pro Ala Thr Ala Pro Ala Asp Gly Asp Ala Gly
740 745 750
Val Leu Ser Glu Phe Leu Ala Val Ala Ser Ala Val Ala Arg Ile Arg
755 760 765
Pro Asp Ala Glu Leu Thr Val Leu Thr Arg Gly Ala Val Pro Ala Ala
770 775 780
Pro Gly Asp Thr Val Val Pro Gly Arg Thr Gly Leu Thr Ala Leu Val
785 790 795 800
Arg Thr Ala Arg Ser Glu Arg Leu Leu Ala Arg Thr Ala Leu Ile Asp
805 810 815
Ala Pro Ala Asp Ala Gly Ala Ala Glu Leu Ala Ala Leu Ala Thr Val
820 825 830
Arg His Ser Gly Asp Val Ile Ala Val Arg Asp Gly Arg Pro Leu Gln
835 840 845
Gln Arg Leu Arg Ala Val Pro Leu Pro Glu Val Phe Asp Val Pro Ala
850 855 860
Asp Val Leu Pro Arg Gly Gly Thr Cys Leu Ile Thr Gly Gly Leu Gly
865 870 875 880
Gly Leu Gly Arg Ala Val Ala Glu His Leu Val Val Thr His Gly Ala
885 890 895
Arg Leu Leu Ile Val Gly Arg Thr Pro Gln Glu Arg Leu Asp Thr Gly
900 905 910
Ala Arg Glu Ala Leu Glu Val Leu His Ala Ile Gly Glu Val Arg Tyr
915 920 925
Arg Thr Ala Asp Ala Ala Asp Pro Asp Ala Leu Ala Gly Ala Val Ala
930 935 940
Glu Ala Glu Ala Ser Trp Gly Cys Gly Leu Asp Leu Val Leu His Leu
945 950 955 960
Ala Gly Ala Ala Val Ala Pro Gln Trp Glu Ser Leu Thr Asp His Glu
965 970 975
Leu Arg Ala Glu Ser Ala Gln Trp Leu Ala Thr Met Leu His Pro Lys
980 985 990
Ala Gly Gly Cys Ala Ala Leu Asp Gly Val Leu Ala Thr Arg Pro Asp
995 1000 1005
Thr Ala Val Val Leu Tyr Ser Ser Val Asn Gly Leu Phe Gly Gly Thr
1010 1015 1020
Gly Phe Gly Ala Tyr Ala Ala Ala Asn Ala Val Leu Asp Gly Trp Ala
1025 1030 1035 1040
Gln Arg Trp Ala Ala Ala Gly Arg Arg Ala Gln Ser Leu Ala Trp Ser
1045 1050 1055
Met Trp Asp Gly Pro Gly Met Asn Gln Gly Ser Pro Leu Val Ala Ala
1060 1065 1070
Gly Arg His Arg Gly Leu Thr Leu Ile Asp Pro Thr Arg Gly Thr Leu
1075 1080 1085
Ala Leu Leu Gly Ala Leu His Ala Gly Pro Val His Leu Leu Ile Gly
1090 1095 1100
Ala Asp Pro Ala Asn Glu Gln Ile Lys Pro Leu Leu Ala Ala Asp Gln
1105 1110 1115 1120
Leu Gly Gly Gly Thr Val Ala Val Ala Val Val Pro Ala Ser Asp Thr
1125 1130 1135
Asp Pro Gln Gln Val Arg Ala Ala Val Ser Ala Gln Leu Ala Asp Arg
1140 1145 1150
Gly Ile Phe Ala Arg Val Val Val Thr Ser Arg Ile Glu Arg Asp Arg
1155 1160 1165
Ser Gly Thr Pro Asp Pro Ala Ala Val Leu Ala Ala Val Gly Ala Gly
1170 1175 1180
Gly Ala Ser Tyr Leu Pro Pro Glu Gly Arg Leu Glu Ala Leu Leu Ala
1185 1190 1195 1200
Glu Ala Ala Ala Glu Val Leu Gly Val Ala Arg Val Gly Arg Asp Asp
1205 1210 1215
Ser Phe Phe Ser Leu Gly Cys Asp Ser Ile Arg Ala Val Gln Leu Ala
1220 1225 1230
Glu Ala Leu Ser Gly Gln Leu Asp Ser Pro Val Ser Val Gly Ser Leu
1235 1240 1245
Tyr Glu Ser Pro Thr Val Arg Ser Leu Ala Ala Arg Ile Ala Gly
1250 1255 1260
<210>3
<211>273
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Leu Ser Thr Asp Thr His Ala Gly Ala Ser Pro Arg Ser Gly Pro Val
1 5 10 15
Ser Phe Thr Met Asp Ser Pro Trp Val Arg Arg Val Pro Arg Glu Gln
20 25 30
Ala Arg Val Arg Leu Ile Cys Leu Pro Phe Ala Gly Gly Gly Ala Ser
35 40 45
Val Tyr Gln Arg Thr Ala Ala Leu Met Pro Ser Trp Val Glu Thr Leu
50 55 60
Thr Val Gln Leu Pro Gly Arg Glu Asp Arg Ser Arg Glu Glu Pro Pro
65 70 75 80
Ala Arg Ile Glu Thr Leu Val Thr Ala Cys Ser Ile Ala Leu Arg Pro
85 90 95
Tyr Thr Thr Met Pro Tyr Val Leu Tyr Gly His Cys Ala Gly Gly Leu
100 105 110
Leu Ala Tyr Glu Ile Ala His Glu Met Gly Arg Arg Phe Gly Thr Trp
115 120 125
Pro Gln Arg Leu Ile Val Gly Glu Gln Pro Ala Pro Gln Ala Pro Pro
130 135 140
Pro Ala Glu Pro Leu His Leu Leu Pro Asp Glu Glu Leu Leu Ala Thr
145 150 155 160
Val Arg Glu Arg Gly Gly Leu Pro Asp Ala Val Ala Arg Asn Ala Pro
165 170 175
Leu Leu Asp Met Leu Leu Pro Leu Leu Arg Ser Asp Phe Glu Leu Trp
180 185 190
Glu Asn Tyr Arg His Arg Pro Arg Pro Pro Leu Pro Val Pro Ile Thr
195 200 205
Thr Val Arg Gly Leu Thr Gly Ser Val Asp Glu Ser Asp Leu Ala Gly
210 215 220
Trp Ala Ala Gln Thr Ala Val Gly Arg Thr Gly Leu Ser Val Glu Gly
225 230 235 240
Gly His Tyr Phe Val Val Gly Leu Thr Pro Glu Ala Ala Arg Glu Ile
245 250 255
Gly Gly Gln Leu Ala Val Glu Pro Pro Ala Pro Ala Gly Arg Thr Ser
260 265 270
Ser
<210>4
<211>1103
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ser Gln Gln Ser Glu Leu Leu Arg Arg Leu Arg Arg Leu Pro Gly
1 5 10 15
Glu Arg Arg Asp Ala Leu Leu Ala Gln Leu Glu Asp Val Ser Ala Gly
20 25 30
Ala Arg Ser Ala Pro Leu Ser Tyr Arg Gln Glu Gln Leu Trp Leu Phe
35 40 45
Asp Arg Phe Ala Pro Ser Ser Leu Ala Tyr Gly Leu Gly Phe Ala Leu
50 55 60
Pro Ile Glu Gly Arg Leu Asp Thr Glu Ala Leu Gly Ala Ala Leu Ala
65 70 75 80
Asp Leu Leu Gln Arg His Thr Val Leu Arg Ser Thr Phe Ser Asp Glu
85 90 95
Asp Thr Arg Gly Thr Gln Arg Leu Lys Pro Val Pro Arg Ile Arg Leu
100 105 110
Glu Thr Glu Asp Val Pro Ala Glu Arg Leu His Pro Arg Ala Asp Glu
115 120 125
Val Val Gln Gly Glu Leu Arg Arg Gly Phe Asp Leu Asn Ser Gly Pro
130 135 140
Met Val Arg Phe Arg Leu Leu Arg Ser Ala Pro Asp Leu His Leu Leu
145 150 155 160
Val Leu Thr Ser His Ser Ile Val Leu Asp Pro Arg Ser Ala Arg Val
165 170 175
Leu Gln Gly Asp Leu Ala His Ala Tyr Arg Ala Arg Ala Ala Gly Ala
180 185 190
Ala Pro Ala Trp Ala Gln Asp Pro Val Glu Phe Gly Thr Tyr Ala Ala
195 200 205
Trp Gln Arg Ala Trp Cys Glu Gln Asp Ala Ala Arg Glu Ala Thr Thr
210 215 220
Tyr Trp Arg Asp Ala Leu Ala Gly Trp Glu Ser Thr Glu Leu Pro Thr
225 230 235 240
Asp Leu Pro Arg Pro Arg Leu Ile Asp Leu Ser Ser Asp Val Val Arg
245 250 255
Arg Pro Leu Pro Pro Gly Leu Ala Ala Ala Ala Asp Asp Leu Ala Thr
260 265 270
Gly Ser Gly Ala Glu Arg Arg Asp Val Leu Leu Ala Ala Phe Leu Val
275 280 285
Val Ile Ala Arg His Thr Ser Arg Gly Asp Leu Thr Ile Gly Leu Pro
290 295 300
Gln Asp Val Ala Gly Pro Tyr Asp Glu Ser Arg Leu Val Gly Asp Ile
305 310 315 320
Gly Asn Leu Leu Pro Leu Arg Val Glu Leu Asp Glu Ala Asp Ser Phe
325 330 335
Ala Ala Val Val Ala Arg Val Arg Asp Ala Arg Arg Asp Gly Leu His
340 345 350
His Ala Ser Leu Pro Phe Lys Leu Ile Leu Asp Ala Leu Gly Val Glu
355 360 365
Pro Asp Ala Gly Arg Leu Pro Leu Val Gln Ile Gly Phe Thr Thr Pro
370 375 380
Val Ala Gly Gly Gly Pro Ala Glu Ala Asp Gly Val Arg Phe Gly Cys
385 390 395 400
Glu Gln Ile Asp Ala Gly Ser Gly Ala Phe Glu Leu Ala Leu Glu Val
405 410 415
Thr Asp Gly Glu Gln Pro Thr Val Ala Val Arg Phe Ala Thr Ser Leu
420 425 430
Tyr Arg Arg Ala Thr Ala Thr Arg Val Leu Asp Arg Phe Leu Gln Thr
435 440 445
Leu Asp Ala Leu Cys Arg Gln Pro Arg Arg Pro Leu Gly Asp Ala Pro
450 455 460
Leu Gly Thr Pro Gln Glu His Ala Asp Val Arg Glu Arg Trp Asn Glu
465 470 475 480
Pro Ile Gly Ala Pro Ala Asp Thr Pro Leu His Asp Leu Phe Ala Glu
485 490 495
Val Val Ala Ala Ala Pro Asp Asn Thr Ala Leu Leu Trp Lys Gly Gly
500 505 510
Ser Leu Ser Tyr Gly Glu Leu Asp Ala Trp Ser Trp Arg Ile Ala Arg
515 520 525
Arg Leu Thr Ala Ala Gly Val Gly Leu Gly Asp Leu Val Pro Val Val
530 535 540
Met Gln Arg Gly Pro Ala Leu Leu Ala Ala Val Leu Gly Val Leu Arg
545 550 555 560
Ala Gly Gly Ala Tyr Val Pro Val Asp Ala Gly Gln Pro Asp Glu Arg
565 570 575
Leu Ala Met Ile Leu Glu Asp Thr Gly Ala Arg Thr Val Val Val Ser
580 585 590
Pro Gly Thr Ala Asp Arg Val Pro Gly Pro Leu Ala Leu Val Val Gly
595 600 605
Ala Asp Pro Leu Asp Gly Pro Gln Ala Glu Glu Glu Pro Pro Ala Val
610 615 620
Val Val Pro Ala Ala Ser Ala Ala Tyr Val Ile Tyr Thr Ser Gly Ser
625 630 635 640
Thr Gly Arg Pro Lys Gly Val Val Val Glu His Arg Asn Ala Ala Asn
645 650 655
Phe Thr Arg Thr Val Arg Asp Met Phe Asp Leu Thr Pro Glu Asp Arg
660 665 670
Val Leu His Phe Ala Ser Pro Gly Phe Asp Val Ser Val Phe Glu Ile
675 680 685
Phe Gly Thr Leu Leu Ser Gly Ala Val Leu Tyr Val Leu Asp Glu Asp
690 695 700
Gln Arg Arg Ser Val Asp Ala Leu Asp Ala Val Leu Gln Glu Gln Arg
705 710 715 720
Ile Thr Val Ile Asp Leu Pro Pro Ala Ile Met Asp Leu Leu Ala Pro
725 730 735
Glu Asn Tyr Pro Asp Leu Arg Val Ala Phe Val Gly Gly Glu Ala Phe
740 745 750
Thr Gly Glu Leu Thr Thr Arg Trp Ala Arg Gly Arg Ser Phe Trp Asn
755 760 765
Gly Tyr Gly Pro Thr Glu Thr Thr Val Thr Val Val Ala Lys Arg Cys
770 775 780
Glu Gly Glu Trp Thr Ala Ser Pro Pro Ile Gly Arg Ala Met Ala Asn
785 790 795 800
His Arg Ala Tyr Val Val Asp Gly Ala Gly Ala Ser Leu Leu Pro Ala
805 810 815
Gly Ala Val Gly Glu Leu Ala Ile Ala Gly Leu Gly Val Ala Arg Gly
820 825 830
Tyr Leu Gly Gln Pro Ala Leu Thr Ala Asp Arg Phe Arg Pro Asp Pro
835 840 845
Asp Gly Pro Pro Gly Ser Arg Arg Tyr Leu Thr Gly Asp Leu Ala Val
850 855 860
Trp Asp Glu Ala Gly Asp Leu Cys Phe Val Gly Arg Ala Asp Arg Gln
865 870 875 880
Val Lys Val Arg Gly Val Arg Ile Glu Leu Gly Glu Val Glu Ala Ala
885 890 895
Leu Gln Ala Val Asp Gly Val Ala Gln Ala Val Ala Glu Val Ala Thr
900 905 910
Asp Pro Arg Thr Gly Ala Leu Leu Val Ala Tyr Val Val Pro Glu Arg
915 920 925
Pro Gly Glu Leu Gln Leu Asp Thr Ile Arg Ser Ala Val Ala Arg Arg
930 935 940
Leu Pro Ala Ala Met Val Pro Ser Val Leu Val Pro Leu Asp Ala Val
945 950 955 960
Pro Leu Thr Pro Ser Gly Lys Ile Asp Arg Arg Arg Leu Pro Ala Val
965 970 975
Glu Phe Ala Val Val Asp Glu Leu Ser Glu Ala Asp Glu Glu Asn Ser
980 985 990
Thr Pro Thr Glu Arg Thr Met Arg Glu Glu Val Phe Thr Pro Leu Leu
995 1000 1005
Gly Ala Arg Ile Gly Asn His Val Asn Phe Phe Ala Ala Gly Gly Thr
1010 1015 1020
Ser Leu Gln Ala Ile Gln Leu Ser Ser Arg Val Asn Ala Val Phe Gly
1025 1030 1035 1040
Val Ala Leu Pro Ile Ala Asp Phe Phe Ala Thr Pro Thr Val Ala Gly
1045 1050 1055
Leu Ala Gly Leu Val Asp Ala Ala Arg Ser Gly Gln Glu Ala Arg Arg
1060 1065 1070
Asp Ala Leu Ala Glu Ala Leu Asp Leu Val Glu Gly Ser Ser Asp Glu
1075 1080 1085
Glu Leu Ala Glu Leu Ala Ala Ser Leu Asp Gly Glu Gly Thr Arg
1090 1095 1100
<210>5
<211>250
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Glu Pro Arg Gln Arg Arg Trp Phe Leu Arg Glu Pro Ser Pro Asp
1 5 10 15
Ala Arg Met Leu Leu Leu Cys Met Pro Tyr Ser Gly Cys Gly Ala Ser
20 25 30
Met Tyr Arg Ala Trp Pro Ala Thr Ile Gly Glu Ala Glu Val Val Pro
35 40 45
Leu Gln Leu Pro Trp Arg Glu Asn Arg Met Arg Glu Pro His Phe Gly
50 55 60
Thr Tyr Glu Glu Leu Ala Ala Ala Leu Leu Asp Asp Val Gly Glu Leu
65 70 75 80
Leu Thr Ser Arg Pro Tyr Ala Leu Phe Gly His Cys Gly Gly Ala Leu
85 90 95
Pro Ala Phe Glu Thr Val Ile Gly Ile Ala Glu Arg Gly Leu Pro Ala
100 105 110
Pro Val Arg Tyr Phe Val Ser Ser Gln Val Ala Pro Gln Asp Gly Pro
115 120 125
Trp Gly Arg Phe Leu Gly Leu Asp Asp Gln Gly Leu Arg Asp Glu Ile
130 135 140
Ala Gly Leu Leu Thr Ala Met Gly Ala Ala Ala Ser Ala Asp Glu Leu
145 150 155 160
Val Asp Leu Phe Ile Asp Val Met His Ala Asp Leu Glu Ala Asn Arg
165 170 175
Lys Tyr Phe Arg Pro Ala Gly Thr Val Pro Cys Pro Ile Thr Ala Leu
180 185 190
Gly Trp Asp Asp Asp Val Glu Val Pro His Arg Leu Met Gly Gly Trp
195 200 205
Gln Ala Trp Ala Pro Thr Asp Lys Val Val Leu Pro Gly Thr His Tyr
210 215 220
Thr Phe Leu Asp Ala Pro Ser Ala Leu Val Arg Val Leu Ala Asp Arg
225 230 235 240
Leu Thr Leu Gly Ser Gly Ser Leu Ala Pro
245 250
<210>6
<211>345
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ala Ala Gly Ser Gly Ser Gly Thr Pro Pro Gly Thr Pro Pro Pro
1 5 10 15
Thr Leu Leu Thr Asp Leu Ala Thr Gly Leu Trp Lys Thr Gln Thr Leu
20 25 30
Thr Ala Ala Ile Glu Thr Gly Leu Phe Glu Ala Leu Ala Ala Gly Asp
35 40 45
Ala Asp Ala Pro Glu Thr Ala Gln Arg Leu Gly Ile Gly Lys Arg Pro
50 55 60
Ala Glu Ile Leu Leu Thr Ala Cys Thr Ala Leu Gly Leu Leu Glu Gln
65 70 75 80
Arg Asp Gly Arg Tyr Arg Asn Thr Ala Val Ala Ala His Tyr Leu Val
85 90 95
Pro Gly Leu Pro Asp Tyr Phe Gly Gly Tyr Val Gln Met Val Ala Arg
100 105 110
Tyr Thr Ala Pro Gly Trp Leu Arg Ala Thr Glu Ala Val Arg Thr Asp
115 120 125
Ala Pro Thr Lys Pro Val Pro Asp Pro Asp Arg Asn Met Phe Glu Glu
130 135 140
Gly Asn Arg Pro Glu Ser Phe Trp Glu Gly Leu Phe Thr Phe Ser Thr
145 150 155 160
Leu Thr Ala Arg Gln Leu Ala Ala Ser Val Asp Leu Ser Gly Val Arg
165 170 175
Arg Ile Met Asp Val Gly Gly Gly Ala Gly Ala Thr Leu Ile Glu Leu
180 185 190
Cys Arg Gln His Pro His Leu Ser Gly Thr Val Val Asp Leu Pro His
195 200 205
Val Cys Ala Leu Ala Gly Glu Arg Ile Ala Ala Ala Gly Met Thr Gly
210 215 220
Arg Ile Asp Thr Ala Ala Ala Asp Phe Phe Ala Asp Pro Leu Pro Ser
225 230 235 240
Gly His Asp Ala Val Leu Leu Ser Met Ile Leu His Asp Trp Asp Glu
245 250 255
Ser Gln Asn Arg Lys Ile Leu Ala Ser Cys Leu Asp Ala Leu Pro Ser
260 265 270
Gly Gly Thr Val Leu Ile Ser Glu Leu Leu Val Asp Asp Asp Lys Ser
275 280 285
Gly Pro Val Asp Ala Ala Leu Met Ser Met Asn Met Leu Val Gly Thr
290 295 300
Trp Gly Arg Asn Tyr Thr Gly Ala Glu Tyr Thr Asp Trp Leu Arg Asp
305 310 315 320
Ala Gly Cys Ser Glu Val Arg Thr Val Arg Phe Ala Ser Pro Gly Ala
325 330 335
Asn Gly Val Val Ala Gly Val Lys Ala
340 345
<210>7
<211>401
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Thr Thr Glu Ala Ala Asp Ala Thr Asp Arg Leu Val Thr Ala Phe
1 5 10 15
Asp His His Asp Pro Gly Tyr Thr Pro Arg Thr Ala Glu Arg Ile Asn
20 25 30
Thr Glu Ile Arg Glu Arg Gly Val Thr Trp Ser Pro Ala Tyr Gly Gly
35 40 45
Ile Trp Ile Leu Ser Arg Tyr Ala Asp Val Arg Ala Ala Leu Thr Asp
50 55 60
Trp Arg Thr Tyr Ser Ser Ala Arg Gly Val His Phe Pro Arg Ala Glu
65 70 75 80
Gly Met Pro Met Phe Ser Pro Ile Asp Tyr Asp Pro Pro Ala Gln Arg
85 90 95
Gly Ile Arg Glu Arg Met Ala Ala Pro Met Thr Gly Asp Ala Val Ser
100 105 110
Ala Met Val Pro Glu Leu Arg Arg Met Val Ala Arg Leu Leu Ala Pro
115 120 125
Leu Ala Gly Arg Gly His Gly Asp Leu Met Ala Glu Phe Ala Glu Pro
130 135 140
Phe Ala Ile Glu Val Leu Gly Val Ala Phe Gly Leu Ser Glu Ser Cys
145 150 155 160
Arg Ala Arg Ile Arg Glu Ala Thr Arg Thr Met Trp Thr Tyr Ile Ser
165 170 175
Ala Asp Arg Asp Ala Ser Lys Phe Trp Pro Ala Phe His Ala Leu Leu
180 185 190
Ala Glu Glu Val Glu Arg Val Arg Asp Glu Pro Asp Gly Ser Tyr Leu
195 200 205
Ala Arg Leu Ala Ala Met Arg Arg Asp Gly Ser Pro Leu Pro Asp Glu
210 215 220
Glu Leu Tyr Ser Ile Ile Val Ser Phe Cys Val Ala Gly His Asp Asn
225 230 235 240
Thr Met Asn Ser Ile Thr Arg Leu Val His Thr Leu Ala Gln Asp Pro
245 250 255
Ala Leu Gln Leu Arg Leu Arg Arg Glu Pro Glu Leu Arg Pro Ala Val
260 265 270
Ala Glu Glu Ala Leu Arg Arg Trp Cys Pro Thr Asp Arg Phe Thr Arg
275 280 285
Val Thr Thr Arg Glu Val Thr Val Ala Gly Thr Val Ile Pro Ala Gly
290 295 300
Ala Arg Val Val Leu Leu Phe Asp Ala Ala Asn Arg Asp Pro Glu Lys
305 310 315 320
Phe Pro Asp Pro Asp Thr Phe Asp Pro Asp Arg Gly Asn Ser His Gln
325 330 335
His Leu Ser Phe Gly His Gly Ile His His Cys Met Gly Val His Leu
340 345 350
Ala Arg Ala Glu Phe Ala Ala Val Leu Asp Glu Leu Ser Arg Leu Pro
355 360 365
Leu Phe Asp Leu Glu Gln Pro Ser Asp Leu His Phe Glu Asn Gly Arg
370 375 380
His Ile Met Phe Asp Arg Val Ser Val Arg Phe Arg Thr Gly Glu Glu
385 390 395 400
His
<210>8
<211>627
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ile Arg Gln Gln Asp Ala Pro Ala Val Pro Gly Ala Gly Pro Ala
1 5 10 15
Thr Thr Val Val Ser Arg Ile Glu Leu Phe Ala Arg Asp Thr Pro Asp
20 25 30
Ala Pro Ala Val Arg His Gly Glu Arg Glu Leu Thr Tyr Arg Asp Leu
35 40 45
Asp Arg Arg Ala Gly Gln Leu Ala Ser Glu Leu Val Glu Ser Gly Val
50 55 60
Arg His Gly Asp Met Val Ala Met Tyr Leu Gly Arg Ser Ile Glu Trp
65 70 75 80
Val Val Gly Met Leu Gly Cys Leu Lys Thr Gly Gly Val Cys Met Pro
85 90 95
Leu Asp Pro Ala Val Pro Ala Glu Arg Phe Asp Arg Ala Val Ala Ala
100 105 110
Ala Arg Pro Ala Ala Val Val Thr Ala Val Pro Gly Arg Gly Thr Gly
115 120 125
Asn Pro Leu Pro Ala Leu Pro Ala Gly Leu Pro Val Val Thr Leu Asp
130 135 140
Ala Ser Ala Pro Leu Pro Ala Glu Thr Pro Gly Gly Gly Leu Ala Pro
145 150 155 160
Arg Ala Gln Pro Arg Cys Glu Asp Leu Ala Tyr Ala Met Phe Thr Ser
165 170 175
Gly Ser Ser Gly Glu Ala Lys Ile Val Leu Ala Gln His Ser Trp Leu
180 185 190
Ala Leu Ser Ala Ala Arg Ser Ala Ala Val Asn Ala Thr Thr Ala Ala
195 200 205
Asp Arg Gly Ser Trp Leu Gly Ala Ala Gly Ala Gly Ile Ala Leu His
210 215 220
Glu Val Gly Gly Leu Leu Trp Gln Gly Ala Gln Leu Val Ile Gly Asp
225 230 235 240
His Glu Val Ile Ala Ser Pro Pro Ala Leu Arg Asp Trp Leu Leu Ala
245 250 255
Glu Arg Ile Thr Gln Ser Phe Val Ile Thr Pro Val Gly Glu Val Leu
260 265 270
Gln Asn Leu Pro Trp Pro Arg Glu Cys Ala Leu Arg Leu Leu Thr Leu
275 280 285
Gly Gly Asp Arg Leu Asn Arg Trp Gly Pro Ala Asp Leu Pro Phe Glu
290 295 300
Val Ala Val Ser Tyr Gly Ser Leu Glu Ala Phe Gln Ile Ala Asn Ser
305 310 315 320
Cys His Pro Trp Ser Gln Arg Cys Thr Pro Ala Thr Ala Thr Asp Ala
325 330 335
Asp Arg Ala Ser Ala Pro Pro Val Gly Arg Pro Ile Ala Gly Val Thr
340 345 350
Val His Leu Leu Glu Asp Asp Gly Leu Thr Pro Val Val Asp Gly Ile
355 360 365
Gly Glu Val Trp Ile Asp Ser Asp Cys Leu Ser Leu Gly Tyr Leu Gly
370 375 380
Asp Pro Ala Gln Thr Ala Asp Arg Phe Arg Pro Asn Pro Phe Gly Ala
385 390 395 400
Ala Gly Ser Arg Ile Tyr Arg Ser Gly Asp Ala Gly Arg Phe Arg Pro
405 410 415
Asp Gly Ile Leu Glu His His Gly Arg Ile Asp Asn Ile Val Lys Ile
420 425 430
Arg Gly His Arg Val Glu Leu Gly Asp Val Glu Trp Ala Leu Gly Gln
435 440 445
His Pro Asp Val Asp Gln Val Ala Val Val Ala Thr Trp Asp Gly Asp
450 455 460
Gln Arg Arg Leu Val Ala Cys Phe Val Ala Ala Arg Glu Ile Ala Pro
465 470 475 480
Leu Glu Leu Arg Asp His Ala Val Glu Arg Leu Pro Asp Trp Met Val
485 490 495
Pro Val Ala Tyr Val Gln Leu Asp Glu Phe Pro Leu Asn Thr Ser Arg
500 505 510
Lys Ile Asp Arg Arg Arg Leu Pro Pro Ala Asp Trp Ser Arg Gly Arg
515 520 525
Pro Ser Arg Pro Trp Arg Ala Pro Ala Pro Gly Ala Gln Thr Glu Leu
530 535 540
Ala Arg Leu Phe Ala Asp Leu Leu Gln Val Asp Arg Val Gly Ala Asp
545 550 555 560
Asp His Phe Val Glu Leu Gly Gly Asp Ser Leu Leu Leu Ala Arg Leu
565 570 575
Gln Ser Arg Ile Glu Gln Arg Phe Gly Val Arg Ile Glu Leu Pro Asp
580 585 590
Leu Met Ala Gly Pro Thr Val Val Glu Leu Ala Gly Arg Ile Pro Gly
595 600 605
Ala Thr Ala Gly Arg Thr Gly Ala Ala Ala Leu Pro Pro Ile Val Pro
610 615 620
Arg Arg Arg
625
<210>9
<211>466
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Thr Ser Thr Asn Ser Ser Ser Gly Thr Gly Thr Thr Pro Ala Ala
1 5 10 15
Ser Glu Ser Ala Gly Ala Glu Val Ser Pro Thr Ser Pro Pro Glu Pro
20 25 30
Ser Gly Asp Thr Ser Ala Pro Thr Gly Gly Thr Ser Thr Lys Pro Arg
35 40 45
Ser Val Leu Phe Arg Asn Arg Asp Phe Ala Leu Val Trp Gly Gly Glu
50 55 60
Ser Ile Ser Leu Leu Gly Ser Glu Ile Ser Val Val Ala Leu Pro Leu
65 70 75 80
Leu Ala Val Ile Thr Phe Gly Glu Gly Ala Phe Gly Val Gly Met Leu
85 90 95
Ile Ala Ala Gln Trp Val Pro Phe Val Val Leu Ala Pro Leu Met Gly
100 105 110
Val Phe Thr Asp Arg Leu Asn Arg Arg Met Leu Met Gln Leu Ser Asn
115 120 125
Ala Val Arg Phe Val Val Leu Gly Ser Leu Pro Leu Ala Trp Ala Leu
130 135 140
Gly Asn Leu Thr Met Val His Leu Tyr Ala Ala Ala Leu Val Lys Gly
145 150 155 160
Val Phe Asp Val Val Phe Gln Leu Ala Tyr Gln Ala Tyr Leu Pro Gln
165 170 175
Leu Leu Ala Arg Glu Asp Leu Val Asp Gly Asn Ala Lys Thr Gln Leu
180 185 190
Ser Arg Ser Val Ala Leu Ile Phe Gly Arg Ser Gly Gly Gly Ala Leu
195 200 205
Val Ser Ala Leu Gly Ala Ala Arg Ala Ile Thr Val Asp Ala Val Ser
210 215 220
Tyr Leu Leu Ala Ala Ile Ala Leu Gly Phe Val Arg Phe Arg Glu Pro
225 230 235 240
Glu Lys Gln Gln Thr Ala Arg Gly Leu Ser Ala Thr Leu Ala Asp Leu
245 250 255
Arg Gly Gly Ile Thr Met Thr Phe Gly Asn Arg Leu Leu Arg Tyr Leu
260 265 270
Thr Leu Met Ala Thr Phe Gly Asn Thr Ala Val Ser Met Val Leu Ala
275 280 285
Met Ile Ile Val Phe Gly Tyr Asp Asp Leu Gly Leu Ser Ala Ala Gln
290 295 300
Val Gly Leu Thr Leu Gly Val Gly Gly Ala Pro Val Val Val Gly Ala
305 310 315 320
Ile Leu Ser Arg Arg Ile Asn Glu Arg Leu Gly Ile Gly Arg Thr Leu
325 330 335
Leu Met Thr His Ser Met Leu Val Val Ala Phe Ala Leu Leu Pro Val
340 345 350
Ser Gln Ala Gly Gly Thr Gly Phe Ala Val Ala Val Leu Ile Val Ser
355 360 365
Gln Gly Ile Ser Ser Phe Thr Thr Pro Val Gly Asn Val Gly Val Met
370 375 380
Thr Leu Ile Gln Lys Ala Thr Pro Pro Gln Ala Met Gly Arg Val Gly
385 390 395 400
Gly Val Ser Leu Pro Leu Val Trp Gly Ala Asn Ala Val Gly Pro Leu
405 410 415
Leu Gly Ser Ala Val Ala Val Thr Ala Gly Asn Ser Ala Ala Phe Tyr
420 425 430
Leu Ala Ser Ala Leu Ala Leu Cys Ala Val Leu Trp Ile Leu Ala Gly
435 440 445
Ser Val Thr Arg Leu Thr Asp Asp Val Pro Glu Asp Met Arg Leu Val
450 455 460
Val Gly
465
<210>10
<211>1115
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Pro Leu Ala Val Ile Gln Glu His Phe Trp Ser Ser Asp Gly Ser
1 5 10 15
Ala Thr Glu Gln Ala Pro Leu Thr Glu Cys Val Ala Leu Ala Val Arg
20 25 30
Gly Pro Leu Asp Ala Ala Ala Leu Arg Thr Ala Val Gly Ala Leu Leu
35 40 45
Ser Arg His Glu Ile Leu Arg Ser Ala Val Arg Leu His Asp Gly Gln
50 55 60
Pro Ser Gln Val Val Leu Pro Val Pro Glu Arg Leu Pro Leu Thr Ile
65 70 75 80
Thr Thr Leu Pro Ser Pro Asp Arg Asp Ala Asp Ala Arg Gln Ala Arg
85 90 95
Asp Arg Glu Leu Ser Arg Phe Ala Ala Thr Gly Ile Asp Pro Ala Ala
100 105 110
Gly Cys Gly Ile Ala Phe Leu Leu Leu Arg Asp Ser Asp Pro Ala Gly
115 120 125
Glu Asp Thr Leu Ala Leu Ala Val His His Ile Phe Ala Asp Ala Thr
130 135 140
Ala Val Arg Leu Leu Leu Ser Glu Leu Ala Ala Asp Tyr Asp Ala Ala
145 150 155 160
Leu Ala Gly Ala Pro Ser Pro Val Pro Glu Pro Glu Leu Gln Tyr Gly
165 170 175
Asp Phe Thr Gln Trp Glu His Ala Ser Leu Leu Pro Ala Val Gln Glu
180 185 190
Pro Asp Thr Ala Trp Trp Arg Glu Thr Leu Arg Asp Ala Pro Thr Ala
195 200 205
Leu Asp Val Arg Pro Asp Arg Pro Arg Arg Arg Val Arg Arg Gly Ala
210 215 220
Gly Arg Arg Val Gly Leu Val Leu Arg Gly Val Asp Gly Pro Ala Val
225 230 235 240
Arg Asn Thr Ala Arg Thr Leu His Ala Ser Pro Tyr Ala Leu Cys Leu
245 250 255
Ala Gly Trp Ala Ala Met Ile Thr Arg Ser Thr Gly Asp Thr Asp Leu
260 265 270
Ile Leu Gly Val Leu Ala Ala Asn Arg Thr Val Pro Gln Leu Glu Pro
275 280 285
Leu Val Gly Gln Phe Ala Asn Thr Val Pro Leu Arg Leu Asp Leu Ala
290 295 300
Gly Ala Pro Asp Leu Ala Asp Ala Val Ser Arg Cys Ser Ala Ala Val
305 310 315 320
Ala Ala Ala Val Glu His Gly Arg Leu Pro Phe Asn Arg Ile Val Arg
325 330 335
Ala Ala Ala Ala Pro Arg Pro Thr Asp Arg Pro Pro Leu Ile Gln His
340 345 350
Met Phe Met Pro Arg Val Asp Ala Val Gly Glu Leu Thr Leu Gly Gly
355 360 365
Gln Pro Val Arg Thr Leu Asp Val Glu Arg Asp Arg Gly Arg Phe Asp
370 375 380
Thr Val Ala Glu Ile Asp Val Thr Gly Asp Gln Val Arg Leu Trp Leu
385 390 395 400
Glu Tyr Asp Ser Asp Leu Tyr Thr Pro Asp Gly Ile His Ala Leu Val
405 410 415
Asp Asp Tyr Ala Gly Val Leu Arg Gln Trp Leu Ala Arg Pro Ala Leu
420 425 430
Arg Leu Pro Glu Leu Glu Leu Ser Ala Pro Glu Pro Ala Gly Leu Ala
435 440 445
Leu Asp Asp Arg Glu Pro Ala Asp Gly Lys Pro Ala Glu Glu Leu Arg
450 455 460
Val Asp Leu Pro Gly Gly Gly Ala Val Thr Phe Leu Leu Asp Arg Ser
465 470 475 480
Pro Gly Pro Glu Tyr Arg Pro Val Leu Ala Gly Leu Arg Gly Ala Arg
485 490 495
Ala Glu Asp Leu Glu Leu Arg Pro Gly Gly Ser Val His His Pro Thr
500 505 510
Asp Leu Leu Leu Ala Gly Val Pro Thr Gly Leu Thr Ala Arg Arg Asp
515 520 525
Pro Arg Gly Arg Leu Glu Ile Val Val Glu Thr Pro Ala Pro Val Ala
530 535 540
Asp Ala Arg Pro Gly Thr Gly Gln Asp Arg Leu Leu Asp Leu Val Thr
545 550 555 560
Glu Ile Trp Ala Gln Ala Leu Glu Val Pro Gly Leu Ala Pro Asp Asp
565 570 575
Asp Phe Phe Thr Cys Gly Gly His Ser Leu Leu Ala Thr Ala Leu Val
580 585 590
Ser Glu Met Gln Glu Thr Leu Gly Val Arg Val Arg Val Arg Ala Leu
595 600 605
Phe Glu Asn Pro Thr Pro Ala Glu Leu Thr Ala His Leu Arg Glu Thr
610 615 620
Glu Pro Glu Leu Asp Gly Met Leu Glu Leu Leu Ala Ala Leu Pro Asp
625 630 635 640
Thr Thr Pro Glu Lys Ser Ala Asp Ala Pro Glu Gln His Asp Ala His
645 650 655
Pro Ala Thr Ala Gln Pro Ala Gly Glu Thr Ala Gln Thr Val Asp Thr
660 665 670
Pro Leu Leu Ser Gly Gln Arg Gln Leu Trp Leu Ala Gln Gln Ala Asp
675 680 685
Pro Asp Ala Leu Thr His Thr Ile Pro Leu Ile Leu Asp Leu Ala Gly
690 695 700
Pro Leu Asn Pro Gly Ala Phe Ala Ala Ala Leu Asn Asp Val Val Ala
705 710 715 720
His Gln Pro Gly Leu Arg Ala Thr Phe Val Glu Val Asp Gly Leu Pro
725 730 735
Val Gln Arg Ile Ser Pro His Thr Pro Val Asp Val Pro Val Ile Asp
740 745 750
Leu Thr Asp His Asp Asp Ala Ala Arg Glu Ala Glu Leu Arg Arg Leu
755 760 765
Glu Gln Glu Ile Ala Tyr Thr Gly Phe Asp Leu Thr Arg Gly Pro Leu
770 775 780
Leu Arg Ala Arg Ile Ile Leu Leu Gly Ala Asp Arg Gly Gln Val Gln
785 790 795 800
Leu Leu Phe His His Leu Val Thr Asp Glu Val Ser Gln Thr Leu Leu
805 810 815
Met Gln Glu Leu Ser Thr Ala Tyr Arg Ala Arg Thr Ala Gly Arg Ala
820 825 830
Pro Val Leu Pro Pro His Glu Thr Asp Leu Ala Thr Leu Ala Lys Asp
835 840 845
Glu Gln Asp Ser Leu Ala Gly Ala Glu Gly Glu Arg Leu Arg Arg Phe
850 855 860
Trp Val Arg Glu Leu Thr Gly Ala Ala Pro Leu Arg Leu Pro Thr Asp
865 870 875 880
Arg Pro Arg Gly Asp Arg Ala Gly Phe Arg Gly Glu Phe Leu Glu Arg
885 890 895
Pro Ala Ser Ala Asp Ala Ala Thr Thr Leu Arg Glu Leu Ala Gly Val
900 905 910
Cys Arg Thr Thr Pro Phe Thr Val Phe Cys Ala Ala Val Ala Ile Val
915 920 925
Leu Arg Arg Leu Ser Gly Gln Ser Asp Ile Val Ile Gly Ile Pro Thr
930 935 940
Ala Asn Arg Thr Gln Arg Gly Ala Asp Arg Leu Ile Gly Cys Phe Leu
945 950 955 960
Asn Val Val Pro Val Arg Leu Asp Leu Ser Gly Asn Pro Arg Phe Asp
965 970 975
Glu Leu Val Gln Arg Val Ser Glu Ala Val Leu Arg Ser Tyr Glu His
980 985 990
Gln Gln Leu Pro Phe Ala Glu Ile Val Gln Ala Val Arg Pro Arg Arg
995 1000 1005
Val Pro Gly Thr His Pro Ile Tyr Gln Val Thr Cys Glu Leu Gln Leu
1010 1015 1020
Glu Ser Trp Met Pro Ala Arg Phe Ala Asp Leu Asp Cys Asp Tyr Arg
1025 1030 1035 1040
Phe Val Ser His Gly Thr Ala Arg Tyr Asp Met Ala Phe His Gly Leu
1045 1050 1055
Leu Arg Pro Ser Gly Leu Ser Ala Met Val Glu Leu Asp Thr Asp Leu
1060 1065 1070
Trp Asp Arg Ala Thr Gly Tyr Arg Arg Ile Asp Gln Val Leu Glu Leu
1075 1080 1085
Leu Ala Val Ala Ala Arg Gly Pro Arg Thr Pro Ile Asp Asp Leu Pro
1090 1095 1100
Val Thr Glu Arg Glu Pro Arg Ser Leu Ser Arg
1105 1110 1115
<210>11
<211>435
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ser Gly Ser Thr Gly Arg Trp Ala Gly Pro Ser Ala Thr Ala Leu
1 5 10 15
Leu Arg Phe Ala Thr Ala Gly Ser Val Asp Asp Gly Lys Ser Thr Leu
20 25 30
Val Gly Arg Leu Leu Tyr Asp Ser Lys Ser Val Leu Thr Asp Gln Trp
35 40 45
Glu Ala Val Glu Arg Ala Ser Arg His Arg Gly Gln Asp Ala Pro Asp
50 55 60
Leu Ala Leu Leu Thr Asp Gly Leu Arg Ala Glu Arg Glu Gln Gly Ile
65 70 75 80
Thr Ile Asp Val Ala Tyr Arg Tyr Phe Ala Thr Ala Arg Arg Arg Phe
85 90 95
Ile Leu Ala Asp Thr Pro Gly His Val Gln Tyr Thr Arg Asn Met Val
100 105 110
Thr Gly Ala Ser Thr Ala Glu Leu Ala Leu Val Leu Val Asp Ala Arg
115 120 125
His Gly Val Val Glu Gln Thr Arg Arg His Ala Thr Ile Ala Ala Leu
130 135 140
Leu Arg Val Pro His Leu Val Leu Ala Val Asn Lys Met Asp Leu Ala
145 150 155 160
Gly Tyr Glu Ala Arg Val Phe Ala Arg Ile Ala Glu Glu Phe Thr Ala
165 170 175
Tyr Ala Thr Glu Leu Gly Val Pro Glu Val Thr Ala Ile Pro Ile Ser
180 185 190
Ala Leu Val Gly Asp Asn Val Val Glu Pro Ser Ala Asn Met Asp Trp
195 200 205
Tyr Gly Gly Pro Thr Val Leu Glu His Leu Glu Thr Val Pro Val Ser
210 215 220
His Asp Leu Thr Gly Cys His Ala Arg Leu Pro Val Gln Tyr Val Ile
225 230 235 240
Arg Pro Arg Thr Ala Glu His Pro Asp Tyr Arg Gly Tyr Ala Gly Gln
245 250 255
Ile Ala Ala Gly Ala Phe Arg Val Gly Gln Glu Val Thr Val Leu Pro
260 265 270
Ser Gly Ala Thr Ser Arg Ile Ser Gly Ile Asp Val Leu Gly Glu Ala
275 280 285
Val Gly Glu Ala Arg Ala Pro Gln Ser Val Thr Leu Leu Leu Glu Asp
290 295 300
Asp Ile Asp Val Ala Arg Gly Asp Leu Leu Val Pro Ser Gly Asp Gly
305 310 315 320
Pro Val Ala Arg Arg Asp Val Glu Ala Thr Val Cys His Val Ala Asp
325 330 335
Ala Pro Leu Ala Val Gly Gln Arg Val Leu Leu Lys His Thr Thr Arg
340 345 350
Thr Val Arg Ala Ile Val Ala Glu Ile Arg Ser Arg Leu Val Leu Asp
355 360 365
Gly Ile Val Ala His Pro Glu Pro Gly Gln Leu Val Ala Asn Asp Ile
370 375 380
Gly Arg Val Arg Ile Arg Thr Ala Glu Pro Leu Ala Val Asp Ala Tyr
385 390 395 400
Ala Asp Ser Arg Arg Thr Gly Ser Phe Leu Leu Ile Asp Pro Ala Asp
405 410 415
Gly Thr Thr Leu Ala Ala Gly Met Ala Gly Pro Ala Phe Gly Ala Ala
420 425 430
Val Thr Gly
435
<210>12
<211>270
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val Ile Leu Phe Ser Gly Gly Lys Asp Ser Ile Val Met Leu His Leu
1 5 10 15
Ala Leu Lys Ala Phe Ala Pro Ala Pro Val Pro Phe Ser Leu Leu His
20 25 30
Val Asp Thr Gly His Asn Phe Pro Glu Val Leu Ala Tyr Arg Asp Arg
35 40 45
Ala Val Ala Arg His Gly Leu Arg Leu His Val Ala Ser Val Gln Asp
50 55 60
Tyr Ile Asp Arg Gly Val Leu Lys Glu Arg Pro Asp Gly Thr Arg Asn
65 70 75 80
Pro Leu Gln Thr Val Pro Leu Thr Glu Lys Ile His Ser Glu Arg Phe
85 90 95
Asp Ala Val Leu Gly Gly Gly Arg Arg Asp Glu Glu Lys Ala Arg Ala
100 105 110
Lys Glu Arg Val Phe Ser Leu Arg Asp Glu Phe Ser Gln Trp Asp Pro
115 120 125
Arg Arg Gln Arg Pro Glu Leu Trp Gln Leu Tyr Asn Gly Arg His Gly
130 135 140
Pro Gly Glu His Val Arg Val Phe Pro Leu Ser Asn Trp Thr Glu Leu
145 150 155 160
Asp Val Trp Gln Tyr Ile Ala Arg Glu Asp Ile Glu Leu Pro Gly Ile
165 170 175
Tyr Tyr Ala His Glu Arg Pro Val Phe Arg Arg Asn Gly Met Trp Leu
180 185 190
Thr Ala Gly Glu Trp Gly Gly Pro Lys Asp Gly Glu Arg Val Glu Thr
195 200 205
Arg Leu Val Arg Tyr Arg Thr Val Gly Asp Met Ser Cys Thr Gly Ala
210 215 220
Val Asp Ser Asp Ala Val Thr Leu Asp Gln Val Val Ala Glu Ile Ala
225 230 235 240
Thr Ser Arg Leu Thr Glu Arg Gly Ala Thr Arg Ala Asp Asp Arg Leu
245 250 255
Ser Glu Ala Ala Met Glu Asp Arg Lys Arg Glu Gly Tyr Phe
260 265 270
<210>13
<211>185
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>l
Val Ile Gly Pro Glu Ser Cys Asp Cys Gly Val Thr Val Leu Leu Thr
1 5 10 15
Gly Leu Pro Ser Ala Gly Lys Thr Thr Val Ala Phe Leu Val Ala Gly
20 25 30
Arg Leu Arg Asp Ala Gly His Arg Val Glu Val Leu Asp Gly Asp Val
35 40 45
Met Arg Lys Thr Leu Ser Ala Gly Leu Gly Phe Gly Arg Ala Asp Arg
50 55 60
His Glu Asn Leu Gln Arg Ile Gly Val Met Ala Glu Val Leu Ala Arg
65 70 75 80
Asn Gly Val Ile Val Leu Val Pro Val Val Ala Pro Tyr Ala Asp Ser
85 90 95
Arg Asp Ala Val Arg Gln Arg His Thr Ala Ser Gly Thr Ala Phe Leu
100 105 110
Glu Val His Val Ala Thr Pro Val Glu Val Cys Ala Gln Arg Asp Val
115 120 125
Lys Gly Leu Tyr Ala Arg Gln Arg Ala Gly Glu Leu Thr Gly Leu Thr
130 135 140
Gly Val Asp Asp Pro Tyr Gln Ala Pro Glu Lys Pro Asp Leu Val Ile
145 150 155 160
Pro Thr Val Arg Gln Ser Ala Asp Ala Ser Ala Ser Leu Leu Met Asp
165 170 175
Leu Leu Val Arg Lys Arg Phe Val Ala
180 185
<210>14
<211>133
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Thr Thr Thr Asp Lys Thr Asp Ser Arg Leu Gly Pro Tyr Val Glu
1 5 10 15
His Leu Gly Leu Gln Phe Glu Arg Ile Asp Pro Asp Arg Ala Val Ala
20 25 30
Tyr Trp Ser Val Arg Ala Asp Leu Leu Gln Pro His Gly Ile Leu His
35 40 45
Gly Gly Val His Cys Ala Val Val Glu Ser Val Ala Ser Ala Ala Ala
50 55 60
Asp Arg Trp Leu Gly Asp Arg Gly Thr Val Val Gly Val Ser Asn Ser
65 70 75 80
Thr Asp Phe Phe Ala Pro Ala Thr Val Ala Asp Gly Arg Leu Thr Ser
85 90 95
Thr Ala Leu Pro Val His Arg Gly Ala Thr Gln Gln Val Trp Ser Val
100 105 110
Glu Thr Val Asp Ala Ala Gly Arg Leu Val Ala Arg Gly Gln Val Arg
115 120 125
Leu His Asn Leu Arg
130
<210>15
<211>62
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Leu Leu Arg Val Glu Arg Gly His Ala Glu Pro Glu Glu Leu Ala Ala
1 5 10 15
Leu Thr Ala Leu Leu Leu Val Cys Ala Ala Ala Arg Arg Gln Ala Gly
20 25 30
Gly Gly Val Pro Ser Arg Asp His Ser Pro Ala Gly Arg Arg Arg Pro
35 40 45
Leu Arg Ile Pro Gln Phe Arg Ala Pro His Ser Trp Gln Gly
50 55 60
<210>16
<211>403
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Thr Ile Gln Arg Pro Ala Ala Ala Pro Gln Leu Ser Val Ala Thr
1 5 10 15
Pro Asp His Arg Val His Arg Leu Pro Ser Leu Thr Gly Leu Arg Phe
20 25 30
Phe Ala Ala Phe Leu Val Val Ile Ser His Val Gly Thr Asn Leu Leu
35 40 45
Pro Arg Val Ala Pro Asp Gln Thr Phe Ala Ile Arg Val Leu Tyr Glu
50 55 60
Cys Gly Ala Phe Gly Val Ser Phe Phe Phe Ile Leu Ser Gly Phe Val
65 70 75 80
Leu Thr Trp Val Ala Arg Asp Glu Asp Ser Val Gly Arg Phe Trp Arg
85 90 95
Arg Arg Phe Phe Lys Ile Tyr Pro Asn His Leu Val Thr Leu Leu Ala
100 105 110
Ala Leu Leu Leu Ala Ala Gly Ala Gly His Ala Leu Ser Gly Arg Asp
115 120 125
Ala Ser Thr Thr Leu Leu Leu Val Gln Ser Trp Ile Pro Asp Met Lys
130 135 140
Leu Gln Tyr Asn Leu Trp Ser Asn Thr Pro Thr Trp Ser Leu Ala Cys
145 150 155 160
Glu Val Leu Phe Tyr Leu Ala Phe Pro Trp Leu Leu Lys Leu Leu Arg
165 170 175
Lys Ile Pro Pro Ala Arg Leu Trp Val Thr Phe Ala Val Val Tyr Ala
180 185 190
Ala Ile Trp Ala Val Pro Leu Val Ala Ser Trp Met Pro Thr Ser Gly
195 200 205
Gly Val His Pro Gly Thr Gly Gln Asn Trp Leu Ser Met Trp Phe Met
210 215 220
Thr Phe Phe Pro Pro Val Arg Met Leu Glu Phe Val Leu Gly Ile Val
225 230 235 240
Thr Ala Leu Ile Val Val Asn Arg Arg Trp Ile Arg Leu Pro Phe Ala
245 250 255
Val Ser Leu Val Phe Pro Leu Ile Pro Leu Val Ala Glu Gly Ser Met
260 265 270
Pro Asp Glu Leu Gly Phe Val Ala Leu Thr Ser Leu Pro Leu Ala Phe
275 280 285
Leu Val Ala Ala Val Ala Ser Ala Asp Ile Asn Gly Arg Asn Gly Arg
290 295 300
Thr Trp Leu Gly Ser Arg Pro Leu Val Phe Leu Gly Glu Ile Ser Phe
305 310 315 320
Ala Leu Tyr Leu Val His Trp Leu Val Val Ala Tyr Gly Trp Ile Gly
325 330 335
Arg Ser Ser Pro Ala Trp Gly Ala Arg Pro Glu Pro Ser Thr Trp Pro
340 345 350
Glu Ile Leu Gly Leu Ala Gly Leu Thr Ile Ala Ser Ser Leu Val Leu
355 360 365
Ala Trp Leu Leu Tyr Thr Leu Val Glu Arg Pro Val Met Arg Arg Trp
370 375 380
Ser Arg Pro Arg Ala Ala Arg Thr Glu Gly Val Leu Thr Ala Ala Pro
385 390 395 400
Glu Arg Gly
<210>17
<211>87
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ala Ser Pro Gly Leu Val His Asp Glu Asp Arg Ile Arg Tyr Val
1 5 10 15
Gln Ser Val Phe Arg Arg Val Leu Arg Leu Glu Gln Asp Ala Ala Pro
20 25 30
Asp Asp Asp Phe Phe Val Leu Gly Gly Thr Ser Leu Ala Ala Met Glu
35 40 45
Val Ile Asp Leu Ile Glu Arg Glu Arg Gly Leu Arg Ile Pro Val Arg
50 55 60
Asn Phe Tyr Arg Ser Thr Val Val Arg Ala Leu Ala Leu Glu Leu Asp
65 70 75 80
Pro Ser Arg Arg Glu Ala Pro
85
<210>18
<211>340
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val Thr Thr Ala Arg Pro Ala Glu Gln Thr Pro Gln Pro Val Leu Gly
1 5 10 15
Pro Asp Pro Arg Ala Ala Tyr Arg Glu Ala Leu Leu Glu Leu Ala Ala
20 25 30
Ala Asp Ser Arg Val Val Cys Leu Asp Ser Asp Thr Gly Gly Leu Glu
35 40 45
Asn Thr Phe Gly Glu Arg Phe Pro Asp Arg Tyr Val Asn Val Gly Ile
50 55 60
Ala Glu Ala Asn Leu Met Thr Val Ala Ala Gly Leu Ala Arg Arg Gly
65 70 75 80
Phe Leu Pro Tyr Val His Thr Met Ala Thr Phe Ala Thr Met Arg Ala
85 90 95
Gly Glu Phe Leu Lys Leu Asp Val Val Gly Asn Arg Leu Pro Val Arg
100 105 110
Val Val Ala Thr His Gly Gly Leu Ser Ala Ala His Phe Gly Thr Ser
115 120 125
His Phe Ala Leu Glu Asp Leu Ala Val Thr Arg Ala Leu Ser Asp Leu
130 135 140
Thr Val Val Val Pro Gly Asp Ala Arg Gln Ile Gly Pro Ala Met Arg
145 150 155 160
Gln Leu His Glu Val Pro Gly Pro Ala Tyr Leu Arg Leu Gly Arg Ser
165 170 175
Ala Thr Pro Val Pro Pro Gly Pro Ala Pro Ala Phe Arg Leu Gly Arg
180 185 190
Ala Arg Val Leu Arg Glu Gly Ala Asp Val Thr Leu Val Ala Ala Gly
195 200 205
Pro Leu Pro Leu Leu Leu Ser Leu Glu Ala Ala Ala Glu Leu Ala Ala
210 215 220
Asp Gly Thr Glu Ala Gln Val Leu Asp Leu His Thr Leu His Pro Leu
225 230 235 240
Asp His Glu Gly Leu Leu Ala Ala Cys Arg Gly Arg Ala Gly Val Val
245 250 255
Thr Val Glu Glu His Arg Pro Gln Gly Gly Leu Gly Asp Ala Val Thr
260 265 270
Glu Val Val Ala Ala Glu Leu Ala Leu Pro Val Arg Arg Val Ala Val
275 280 285
Arg Gly Arg Pro Gly Val Arg Val Ala Gly Gln Arg Ala Ala Leu Glu
290 295 300
Gln Leu Gly Val Ser Thr Gly Ala Val Ala Asp Ala Ala Arg His Leu
305 310 315 320
His Ala Ala Arg Ala Glu Ser Pro Ala Gln His Thr Ala Ala Gly Pro
325 330 335
Thr Met Arg Gly
340
<210>19
<211>311
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val Ser Arg Pro Pro Leu Pro Arg Arg Phe Leu Arg Ser Thr Ser Trp
1 5 10 15
Ser Thr Ser Arg Thr Asp Ala Arg Pro Lys Gly Ile Arg Thr Met Ala
20 25 30
Val Pro Leu Thr Gly Pro Pro Gln Pro Pro Ala Pro Ala Pro Pro Pro
35 40 45
Gln Pro Leu Asp Pro Ala Asp Met Ala Arg Arg Ile Arg Gln Asp Val
50 55 60
Ile Arg Met Ala Gly Gly Pro Glu Gly Ala His Val Gly Gly Ser Leu
65 70 75 80
Ser Cys Ala Asp Val Leu Ala Val Leu His Thr Arg Leu Leu Arg Pro
85 90 95
Gly Asp Ser Phe Val Leu Ser Lys Gly His Ala Ala Pro Ala Leu Tyr
100 105 110
Ser Thr Leu Val Gln Leu Gly Arg Leu Pro Ala Asp Glu Leu Asp Ser
115 120 125
Tyr Thr Ala Pro Gly Ser Arg Leu Phe Gly His Pro Ser His Asp Leu
130 135 140
Pro Gly Val Glu Phe Ala Thr Gly Ser Leu Gly His Gly Leu Gly Leu
145 150 155 160
Ser Val Gly Leu Ala Ile Ala Glu Gln Leu His Gly Gly Asp Gly Arg
165 170 175
Val Tyr Thr Leu Leu Gly Asp Gly Glu Leu Gln Glu Gly Ser Val Trp
180 185 190
Glu Ala Ala Leu Leu Ala Gly His Arg Arg Pro Pro Gly Leu Val Ala
195 200 205
Val Val Asp Arg Asn Gly Leu Gln Ile Thr Gly Gly Thr Glu Asp Cys
210 215 220
Val Gly Leu Glu Pro Leu Asp Ala Lys Phe Ala Ala Phe Gly Trp Glu
225 230 235 240
Thr Arg Thr Val Asp Gly His Asp Leu Pro Ala Leu Tyr Glu Ala Leu
245 250 255
Gln Pro Ser Pro Gly Gly Pro Val Ala Val Leu Ala Arg Thr Val Lys
260 265 270
Gly Arg Gly Val Pro Phe Leu Glu Ala Arg Val Ala Gly His Tyr Ala
275 280 285
Lys Leu Lys Pro Ala Leu Val Ser Arg Ala Leu Ala Ala Leu Gly His
290 295 300
Pro Val Gln Gly Gly Thr Ala
305 310
<210>20
<211>440
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Pro Glu Leu Ala His Ala Thr Thr Arg Leu Ala Leu Leu Gly Gly
1 5 10 15
Asp Arg Ala Val Pro Lys Asp Ser Val Gly Gly Thr Trp Pro Leu Val
20 25 30
Thr Thr Ala Asp Glu Gln Ala Val Leu Arg Val Leu Arg Glu Gly Pro
35 40 45
Leu Asn Ala Ala Ser Pro Gly Glu Arg Glu Val Gly Gly Leu Glu Arg
50 55 60
Ala Trp Ala Gln Ala Thr Gly Thr Arg His Cys Val Ala Val Ala Ser
65 70 75 80
Gly Thr Ala Ala Leu His Leu Ala Leu Arg Gly Cys Gly Ile Gly Pro
85 90 95
Gly Asp Glu Val Ile Val Pro Ala Leu Thr Met Asn Ala Thr Ala His
100 105 110
Ala Val Cys Ala Ala Gly Ala Thr Pro Val Phe Ala Asp Val Asp Pro
115 120 125
Asp Thr Tyr Thr Leu Asp Pro Arg Ser Ala Ala Ala Val Cys Gly Pro
130 135 140
Arg Thr Ala Ala Leu Leu Pro Val His Leu His Gly Leu Pro Ala Asp
145 150 155 160
Met Arg Ala Leu Asp Glu Leu Ala Gly Arg Arg Gly Val Pro Val Val
165 170 175
Glu Asp Ala Ala Gln Ala His Gly Ala Gln Leu Asp Gly Arg Arg Val
180 185 190
Gly Ala Leu Gly Lys Ala Gly Cys Phe Ser Leu His Pro His Lys Asn
195 200 205
Leu Pro Ser Cys Gly Glu Gly Gly Leu Ile Thr Thr Asp Asp Asp Ser
210 215 220
Leu His Glu Gln Leu Val Arg Leu Arg Asn Phe Gly Glu Leu Glu Pro
225 230 235 240
Thr Gly Ala Arg Ser Tyr Ile Ala His Arg Ala Ala Leu Asn Ala Arg
245 250 255
Ile Thr Pro Met Ala Ala Ala Phe Thr Arg Ser Gln Leu Ala Arg Phe
260 265 270
Asp Asp Tyr Ala Arg Arg Arg Glu Pro Gly Ile Arg Arg Phe Leu Asn
275 280 285
Arg Leu Ala Gln Leu Pro Gly Leu Arg Val Pro Gln Val Pro Ala Gly
290 295 300
Arg Thr His Ala Trp His Ile Leu Arg Leu Met Pro Asp Pro Glu Ala
305 310 315 320
Phe Gly Leu Gly Pro Glu His Arg Cys Ala Leu Arg Ala Ala Leu His
325 330 335
Arg Ile Leu Arg Ala Glu Gly Val Pro Val Ser Arg Tyr Gln Val Ala
340 345 350
Pro Leu Ala Ala Gln Pro Ala Phe Arg Pro Pro Gly Ala Ser Pro Ala
355 360 365
Asp Ala Ala Asp Gln Val Asp Ala Leu Leu Pro Val Thr Val Ala Val
370 375 380
Val Asp Gly Ser Leu Cys Leu Gln Arg Arg His Leu Asp Pro Ala Ala
385 390 395 400
Gly Pro Leu Leu Gln Ser Tyr Ala Asp Ala Phe Glu Lys Val Trp Gln
405 410 415
Asn Leu Glu Leu Val Arg Arg Met Ala Thr Ser Arg Ser Ala Pro Gly
420 425 430
Trp Arg Glu Ala Leu Arg Gln Gly
435 440
<210>21
<211>307
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val His Ala His Pro Lys Leu Glu Leu Ala Gln Ala Leu Ser Arg Arg
1 5 10 15
His Ala Gly Arg Pro Val Ala Glu Val His Pro Asn Leu His His Leu
20 25 30
Ala Leu Ala Phe Arg Ser Pro Glu Glu Leu Glu Pro Val Asp Ala Ala
35 40 45
Phe Leu Ala Leu Pro Ser Gly Ala Ser Ala Glu Val Glu Thr Ala Val
50 55 60
Thr Gly Arg Ala Thr Thr Ile Val Asp Leu Ser Pro Asp Phe Arg Gly
65 70 75 80
Gly Arg Pro Ser Ala Asp Gly Thr Ala Tyr Arg Thr Gly Leu Pro Glu
85 90 95
Leu Phe Ala Gly Glu Leu Val Gly Ala Thr Arg Ile Ser Val Pro Gly
100 105 110
Cys Met Ala Thr Ala Ala Val Leu Ala Leu Ala Pro Val Gln Arg Ser
115 120 125
Gly Leu Val Ala Gly Asp Val Val Val Asp Ala Arg Ala Gly Ser Ser
130 135 140
Gly Ser Gly Ala Glu Pro Ser Pro Ala Ser His His Ala Glu Arg Gly
145 150 155 160
Asn Ala Phe Arg Val Tyr Lys Pro Ala Gly His Arg His Glu Lys Glu
165 170 175
Ile Ala Ala Leu Ala Gly Val Arg Ala Arg Met Thr Val Thr Ala Val
180 185 190
Pro Thr Val Arg Gly Ala Gln Val Leu Cys His Val Thr Pro Pro Arg
195 200 205
Pro Val Thr Arg Arg Glu Val Leu His Val Leu Gln Arg Ala Tyr Arg
210 215 220
Asp His Pro Phe Val Arg Ile Val Ser Arg Gly Arg Gly Val His Arg
225 230 235 240
Met Pro Asp Pro Gln Phe Leu Gly Gly Thr Asn Phe Val Asp Ile Gly
245 250 255
Cys Ala Val Asp Glu Asp Gly Gly Arg Ile Val Leu Val Ala Ala Leu
260 265 270
Asp Asn Leu Val Lys Gly Ala Ala Gly Gly Gly Val Gln Ser Leu Asn
275 280 285
Val Ala Ala Gly Met Pro Gln Asp Ala Gly Leu Glu Phe Thr Gly Leu
290 295 300
His Pro Ala
305
<2l0>22
<211>292
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ala Gln Asp Pro Arg Gly His Glu Pro Asn Pro Leu Ile Gly Gly
1 5 10 15
Phe Ala His Asp Gly Pro Ile Val Val Lys Ile Gly Gly Gly Ser Asp
20 25 30
Pro Glu Pro Val Leu Asp Glu Val Ala Glu Leu Ala Leu Ala Gly Arg
35 40 45
Pro Thr Val Leu Val His Gly Gly Gly Ala Val Ala Asp Leu Leu Ser
50 55 60
Glu Gln Leu Gly Val Glu Arg Arg Val Ile Arg Ser Pro Asp Gly Thr
65 70 75 80
His Ser Arg Arg Thr Asp Ala Ala Met Leu Asp Val Ile Thr Leu Ala
85 90 95
Leu Leu Gly Arg Val Lys Pro Arg Leu Ile Ser Gly Leu Arg Ala Arg
100 105 110
Gly Ala Arg Ser Val Gly Leu Ser Gly Ala Asp Gly Ala Leu Leu Thr
115 120 125
Ala Thr Arg Lys Pro Ala Leu Arg Ser Val Gln Asp Gly Arg Thr Val
130 135 140
Leu Ile Arg Asp Asp Arg Ser Gly Arg Ile Glu Arg Val Asp Pro Ala
145 150 155 160
Pro Val Arg Ala Val Leu Asp Arg Gly His Val Pro Val Val Ser Pro
165 170 175
Pro Ala Ser Asp Ala Ala Gly Asn Leu Leu Asn Val Asp Ala Asp Glu
180 185 190
Ala Ala Ala Arg Leu Ala Thr Ala Leu Asp Ala Ser Ala Leu Val Leu
195 200 205
Leu Thr Asp Val Gly Gly Val Leu Ala Asp Leu Gly Asp Pro Ala Thr
210 215 220
Arg Ile Ala Arg Val Gly Pro His His Leu Glu Gly Asp Val Val Arg
225 230 235 240
Gly Arg Met Arg His Lys Val Arg Ala Gly Leu Arg Ala Ser Arg Thr
245 250 255
Val Arg Gln Val Ala Ile Gly Ala Ala His Leu His Arg Pro Ile His
260 265 270
Gln Ala Leu Ser Gly Ala Gly Ser Arg Leu Cys Asp Glu Gly Glu Arg
275 280 285
Pro Glu Asp Gly
290
<210>23
<211>376
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ala Asp His Gln Arg Thr Ala Asp Arg Thr Arg Pro Ala Ala Gly
1 5 10 15
Asp Gly Asp Tyr Pro Ala Trp Leu Leu Arg Ala Ala Leu Gly Ile Pro
20 25 30
Ser Val Ser Gly Arg Glu Leu Pro Val Ala Arg Phe Leu Ala Asp Arg
35 40 45
Met Ala Glu Leu Gly Leu Asp Ser Tyr Val Asp Glu Val Gly Asn Val
50 55 60
His Gly Leu Ile Gly Pro Arg Thr Gly Pro Thr Val Met Leu Leu Gly
65 70 75 80
His Ile Asp Thr Val Pro Gly Gln Val Pro Val Val Gln Val Gly Asp
85 90 95
Leu Leu Tyr Gly Arg Gly Ala Val Asp Ala Lys Gly Ser Ile Val Thr
100 105 110
Met Ile Cys Ala Ala Ala Arg Thr Glu Gly Val Arg Val His Val Ile
115 120 125
Gly Ala Val Gly Glu Glu Val Ala Gly Ser Arg Gly Ala His His Val
130 135 140
Leu Ala Thr Val Pro Pro Pro Ala Ala Leu Val Ile Gly Glu Pro Ser
145 150 155 160
Gly Trp Asp Gly Val Cys Leu Gly Tyr Lys Gly Arg Ile Gly Ile Gly
165 170 175
Tyr Glu Ile Ser Arg Pro Pro Leu His Thr Ser Ser Pro Glu Pro Thr
180 185 190
Ala Val Glu Cys Ala Ala Arg Phe Ala His Asp Ile Gln Glu Tyr Leu
195 200 205
Arg Gly Leu Ser Pro Glu Gln Asp Asp Pro Val Ala Phe Gly Val Ala
210 215 220
Ala Ala Thr Leu Val Arg Leu His Gly Asp Leu Ala Glu Ala Glu Ala
225 230 235 240
Phe Leu Ser Cys Arg Val Pro Pro Gly Phe Asp Phe Ala Ala Leu Glu
245 250 255
Arg Phe Ala Arg Ser Arg Pro His Ser Arg Ile Arg Val Asp Glu Arg
260 265 270
Val Pro Gly Val Thr Arg Pro Arg Thr Asp Pro Val Val Ser Gln Leu
275 280 285
Arg Ala Ala Ile Ala Ala Gln Gly Val Arg Pro Thr Leu Lys Arg Lys
290 295 300
Ala Gly Thr Ser Asp Met Asn Thr Leu Ala Pro Trp Gly Ile Pro Met
305 3l0 315 320
Ala Ala Tyr Gly Pro Gly Asp Ala His Leu Asp His Thr Thr Asp Glu
325 330 335
His Ile Pro Leu Gly Asp Leu Tyr Arg Ala Ile Asp Val Leu Ala Leu
340 345 350
Ala Leu Pro Arg Leu Ala Ser Arg Leu Ala Ala Pro Pro Arg Thr Thr
355 360 365
Gly Ala Pro Glu Leu Leu Glu Ala
370 375
<210>24
<211>272
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Leu Asp Gly Asp Val Ala Leu Ile Thr Gly Ala Ala Arg Ala Leu
1 5 10 15
Gly Arg Ser His Ala Leu Ala Phe Ala Arg Glu Gly Ala Asp Leu Ile
20 25 30
Leu Leu Asp Arg Cys Ala Pro Asp Asp Gly Pro Tyr Pro Met Ala Gly
35 40 45
Arg Arg Asp Leu Glu Glu Thr Ala Glu Ala Cys Arg Arg Leu Gly Ser
50 55 60
Arg Val Leu Thr Gly Val Ala Asp Val Arg His Gln Ala Asp Val Asp
65 70 75 80
Lys Val Val Asp Ala Gly Leu Asp Ala Phe Gly Arg Ile Asp Val Leu
85 90 95
Leu Asn Asn Ala Gly Val Leu Gly Pro Gly Gly Val Arg Val His Glu
100 105 110
Leu Ser Glu Glu Gln Trp Gln Leu Ala Leu Asp Val Asn Leu Ser Gly
115 120 125
Thr Trp Arg Thr Cys Arg Ala Val Leu Pro His Met Val Glu Arg Gly
130 135 140
Gly Gly Arg Ile Val Asn Thr Ala Ser Thr Gly Gly Arg Val Ala Phe
145 150 155 160
Glu Arg Tyr Ala Asn Tyr Val Ala Ser Lys His Gly Val Ile Gly Leu
165 170 175
Thr Lys Ser Ile Ala Leu Glu Tyr Gly Arg His Gly Ile Arg Ala Asn
180 185 190
Ala Val Cys Pro Thr Thr Val Ala Asp Asp Ala Ala Leu Gly Thr Arg
195 200 205
Ser Thr Arg Ala Val Ala Ala Ser Met Gly Ala Glu Leu Pro Asp Tyr
210 215 220
Glu Arg Thr Ser Ala Ala Leu His Pro Ile Gly Arg Leu Val Thr Ala
225 230 235 240
Ala Glu Val Ser Ala Ala Cys Leu Trp Leu Ala Gly Asp Arg Ser Gly
245 250 255
Gly Val Thr Gly Thr Glu Leu Leu Val Asp Gly Gly Phe Thr Ala His
260 265 270
<210>25
<211>1541
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Asp Leu Pro Thr Glu Gln Pro Arg Ser Leu Pro Asp Thr Pro Val
1 5 10 15
Ala Ala Ala Val Pro Pro Asp Leu Ala Ala Leu Arg Ser Arg Leu Ala
20 25 30
Glu Leu Ser Pro Glu Lys Arg Arg Leu Val Glu Arg Leu Leu Ala Arg
35 40 45
Gln Gly Ala Gly Leu Gly Ile Thr Arg Arg Ala Gly Gly Ala Pro Glu
50 55 60
Ala Pro Cys Ser Phe Glu Gln Glu Arg Leu Trp Phe Met Tyr Glu Leu
65 70 75 80
Leu Thr Arg Arg Glu Ile Phe His Val Pro Val Ala Leu Arg Leu Glu
85 90 95
Gly Glu Leu Asp Pro Asp Ala Leu Glu Arg Ala Leu Arg Gln Leu Gly
100 105 110
Arg Arg His Glu Ala Leu Arg Thr Val Phe Arg Gln Arg Asp Gly Arg
115 120 125
Pro Tyr Gln Val Val Arg Glu Gln Leu Asp Leu Pro Leu Glu Arg Val
130 135 140
Asp Cys Arg Thr Ala Ala Asp Pro Ala Leu Ala Ala Arg Gln Gln Ala
145 150 155 160
Ser Ala Leu Val Thr Glu Asp Phe Asp Leu Glu Glu Gly Pro Leu Val
165 170 175
Arg Cys Thr Leu Tyr Arg Thr Gly Glu His Glu His Leu Leu Ala Val
180 185 190
Val Gln His His Ile Val Ser Asp Asn Trp Ser Leu Gly Ile Leu Leu
195 200 205
Asp Asp Leu Gly Lys Leu Tyr Ala Arg Glu Leu Gly Ala Pro Gly Glu
210 215 220
Leu Pro Pro Leu Asp Leu His Tyr Pro Asp Phe Ala Ala Trp Gln Arg
225 230 235 240
Ala Thr Val Asp Ser Thr Thr Met Arg Arg Thr Leu Asp His Trp Arg
245 250 255
Asp Arg Leu Asp Gly Ala Pro Asp Ser Leu Asp Leu Pro Thr Asp Arg
260 265 270
Pro Arg Pro Ala Val Arg Gly Ser Gln Gly Lys Phe His His Val Arg
275 280 285
Phe Asp Ala Asp Leu Val Ala Gly Leu Arg Glu Ala Ala Arg Gln His
290 295 300
Asp Thr Thr Leu Leu Gly Ala Phe Leu Ala Gly Tyr Ile Ala Leu Leu
305 310 315 320
Ser Arg Leu Val Arg Ser Asp Ser Leu Val Val Gly Val Pro Val Ala
325 330 335
Gly Arg Pro Arg Ala Glu Met Gln Gln Met Ile Gly Tyr Phe Leu Asn
340 345 350
Trp Leu Pro Ile His Val Gln Val Gly Asp Arg Pro Asp Leu His Thr
355 360 365
Leu Ile Arg Arg Thr Gly Thr Ala Leu Ser Glu Ala Met Thr His Gln
370 375 380
Asp Val Pro Phe Asp Met Leu Val Arg Glu Leu Arg Pro Ser Arg Arg
385 390 395 400
Pro Gly Glu Thr Pro Ile Phe Gln Thr Ser Phe Ser Leu Arg Asp Gly
405 410 415
Ala Pro Thr Pro Pro Ala Met Pro Gly Ile Asp Val Thr Phe Ala Glu
420 425 430
Leu Asp Gly Gly Ala Thr His Tyr Asp Leu Met Ala Glu Leu Trp Cys
435 440 445
Glu Gly Asp Glu Val Val Gly Tyr Val Pro Tyr Asp Asp Glu Leu Leu
450 455 460
Asp Ala Gln Thr Val Ala Arg Trp Ala Gly Trp Leu Lys Thr Leu Leu
465 470 475 480
Arg Ala Gly Leu Ala Ala Pro Asp Thr Pro Val Ala Asp Leu Glu Met
485 490 495
Leu Ala Pro Gly Glu Ser Val Val Ile Pro Ala Arg Ala Pro Val Thr
500 505 510
Ala Glu Gly Gly Val Ala Arg Pro Ala Arg Thr Leu His Gly Val Phe
515 520 525
Ala Ala Gln Ala Ala Arg Arg Pro Asp Ala Pro Ala Val Ser Asp Glu
530 535 540
Arg Asp Arg Leu Thr Tyr Ala Glu Leu Ser Ala Arg Ala Asp Arg Ile
545 550 555 560
Ala Ala Ala Leu Gln Glu Arg Gly Glu Gly Pro Gly Ser Ile Val Gly
565 570 575
Leu Val Leu Asp Arg Thr Val Asp Leu Pro Ala Ala Val Leu Gly Val
580 585 590
Leu Arg Ala Gly Ala Ala Tyr Leu Pro Val Asp Pro Glu Asn Pro Ala
595 600 605
Gly Arg Thr Ala Asp Gln Phe Thr Glu Cys Arg Val Arg Thr Val Leu
610 615 620
Thr Thr Pro Ala Thr Ala Ser Ala Pro Ala Leu Glu Glu Leu Asp Asp
625 630 635 640
Gly Pro Arg Ile Leu Val Val Asp Trp Gln Asp Pro Gly Trp Arg Glu
645 650 655
Gln Thr Pro Gly Pro Ala Ala Val Asp Val Pro Ala Asp Ala Pro Ala
660 665 670
Tyr Val Ile Tyr Thr Ser Gly Ser Thr Gly Arg Pro Lys Gly Val Leu
675 680 685
Val Thr His Arg Asn Val Leu Arg Leu Phe Thr Ala Cys Asp Glu His
690 695 700
Leu Arg Val Gly Pro Asp Asp Val Trp Thr Leu Phe His Ser Tyr Ala
705 710 715 720
Phe Asp Phe Ser Val Trp Glu Ile Trp Gly Ala Leu Leu His Gly Gly
725 730 735
Arg Leu Val Val Val Pro Gln Trp Ala Thr Arg Ala Pro Asp Val Phe
740 745 750
Ala Glu Leu Val Arg Asp Glu Arg Val Thr Val Leu Ser Gln Thr Pro
755 760 765
Ser Ala Phe Gly Gln Val Ser Glu Thr Leu Leu Lys His Pro Asp Pro
770 775 780
Ala Ala Leu Arg Tyr Val Val Phe Gly Gly Glu Ala Leu Asp Pro Thr
785 790 795 800
Ala Leu Arg Pro Trp Thr Arg Val Tyr Gly Asp Arg Arg Pro Glu Leu
805 810 815
Ile Asn Met Tyr Gly Ile Thr Glu Thr Thr Val His Val Thr Ala Arg
820 825 830
Pro Leu Arg Glu Lys Asp Leu Ala Gly Thr Ala Ser Pro Ile Gly Pro
835 840 845
Pro Leu Ala Asp Leu Ser Leu Tyr Leu Leu Asp Asp Ser Leu Arg Pro
850 855 860
Val Gly Thr Gly Val Pro Gly Glu Ile Phe Val Gly Gly Asp Gly Val
865 870 875 880
Ser Leu Gly Tyr Val Ala Asn Pro Gly Leu Thr Ala Glu Arg Met Leu
885 890 895
Pro Asp Pro Phe Ala Gly Arg Pro Gly Ala Arg Met Tyr Arg Ser Gly
900 905 910
Asp Leu Ala Val Arg Arg Gly Asp Gly Glu Leu Val Cys Leu Gly Arg
915 920 925
Ala Asp Asp Gln Val Lys Ile Arg Gly His Arg Ile Glu Pro Gly Glu
930 935 940
Val Arg Ala Ala Leu Asp Ala Leu Ala Val Val Ala Arg Ala Ala Val
945 950 955 960
Val Val Glu Arg Asp Arg Val Gly Ala Ala Val Leu Ala Ala Tyr Val
965 970 975
Val Pro Ala Glu Gly Asn Ala Gly Val Ser Gly Thr Gly Ile Arg Arg
980 985 990
Thr Leu Leu Arg Ser Leu Pro Glu Trp Met Val Pro Ala Ser Val Thr
995 1000 1005
Val Leu Asp Glu Leu Pro Leu Thr Arg Asn Gly Lys Leu Asp Arg Arg
1010 1015 1020
Ala Leu Thr Asp Arg Lys Glu His Ala Ala Pro Ala Gly Pro Arg Gly
1025 1030 1035 1040
Glu Ala Pro Arg Ser Asp Thr Ala Arg Gln Leu Ala Glu Ile Trp Gln
1045 1050 1055
Glu Leu Leu Gly Val Pro Ala Val Gly Gly Glu Asp Ser Phe Phe Glu
1060 1065 1070
Leu Gly Gly His Ser Leu Met Val Met His Leu Val Ala Arg Ile Arg
1075 1080 1085
Thr Ala Phe Gly Val Glu Met Pro Val Glu Thr Leu Phe Arg Arg Pro
1090 1095 1100
Gln Leu Gln Pro Leu Ala Asp Glu Val Asp Ala Ala Arg Thr Ala Ala
1105 1110 1115 1120
Gln Arg Ala Ala Ala Pro Glu Pro Gly Gln Asp Ala Ala Ala Glu Leu
1125 1130 1135
Ala Asp Val Arg Ala Asp Leu Ala Ala Arg Ala Ala Gly Ile Pro Arg
1140 1145 1150
Pro Gly Ala Arg Ala Asp Ala Asp Arg Asp Thr Val Leu Leu Thr Gly
1155 1160 1165
Ala Thr Gly Phe Val Gly Arg Phe Val Leu Ala Glu Leu Leu Ala Ala
1170 1175 1180
Gly Ala Arg Val Ile Cys Leu Leu Arg Gly Gly Thr Ala Arg Arg Glu
1185 1190 1195 1200
Glu Leu Val Ala Gly Met Ala Asp Leu Gly Leu Trp His Glu Glu His
1205 1210 1215
Ala ALa Arg Leu Glu Leu Val Asp Gly Asp Ile Ala Glu Pro Gly Leu
1220 1225 1230
Gly Leu Ala Gly Pro Asp Arg Asp Arg Leu Ala Asp Arg Ala Gly Arg
1235 1240 1245
Ile Ile His Ala Ala Ala Trp Val Asn His Val Tyr Pro Tyr Glu Arg
1250 1255 1260
Leu Ala Ala Ala Asn Thr His Cys Met Ala Gly Leu Leu Glu Leu Ala
1265 1270 1275 1280
Ala Arg Gly Arg Arg Ser Ala Leu Thr Val Val Ser Thr Ser Ser Val
1285 1290 1295
Ala Asp Ser Ala Ala Tyr Pro Pro Gly Ser Thr Val Pro Pro Gly Pro
1300 1305 1310
Leu Lys Ala Leu Pro Ser Ala Ala Asn Gly Tyr Val Arg Ser Lys Ala
1315 1320 1325
Val Ala Glu Gln Tyr Leu His Leu Ala Ala Glu Leu Asp Val Pro Ala
1330 1335 1340
Ala Val Ile Arg Ile Pro Ser Val Phe Gly Asp Gln Arg Arg Tyr Gln
1345 1350 1355 1360
Ile Asn Pro Ala Asp Ala Val Trp Ser Trp Cys Arg Ala Met Ile Glu
1365 1370 1375
Thr Ser Gly Phe Pro Glu Ser Phe Ala Gln Pro Gly Asn Glu Leu Phe
1380 1385 1390
Gln Ala Leu Pro Ala Asp Ala Val Ala Arg Ala Val Leu Leu Ala Asp
1395 1400 1405
Arg Asp His Thr Glu Pro Gly Thr Arg Tyr Leu Asp Ala Val Pro Ala
1410 1415 1420
Ala Val Gly Thr Thr Glu Asp Leu Leu Ala Ala Leu Arg Ala Ala Gly
1425 1430 1435 1440
His Ala Leu Ser Pro Cys Pro Asp His Ala Trp Tyr Thr Ala Val Gly
1445 1450 1455
Glu Leu Asp Pro Gly Arg Val Trp Val Ala Gly Ile Ala Gly Gln Ala
1460 1465 1470
Ala Ala Arg Leu Ala Ala Asp Pro Ser Ala Thr Ala Pro Arg Thr Leu
1475 1480 1485
Arg Arg Phe Thr Ala Pro Val Glu Pro Gly Glu Leu Asp Glu Leu Leu
1490 1495 1500
Arg Thr Arg Ala Leu Tyr Ser Ser Ala Gln Leu Ala Gly Tyr Ile Arg
1505 1510 1515 1520
Thr Leu Asp Ala Ser Phe Pro Pro Ala Gly Asn Asp Arg Glu Lys Ala
1525 1530 1535
Arg Pro Ala Arg Gly
1540
<210>26
<211>1779
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Leu Ala Glu Asn Val Gln Asn Pro Pro Val Glu Pro Leu Ala Val Ile
1 5 10 15
Gly Met Ser Cys Arg Phe Ala Pro Asp Leu Asp Thr Pro Gly Arg Leu
20 25 30
Trp Glu Phe Leu Arg Ala Gly Gly Ser Ala Val Gly Glu Met Pro Asp
35 40 45
Arg Arg Trp Asp Pro Tyr Val Thr Asp Ser Arg Thr Arg Asp Ile Leu
50 55 60
Arg Thr Thr Thr Arg Lys Gly Ser Phe Met Arg Asp Ile Glu Gly Phe
65 70 75 80
Asp Ala Glu Phe Phe Gln Ile Thr Pro Arg Glu Ala Glu Tyr Ile Asp
85 90 95
Pro Gln Gln Arg Ile Met Leu Glu Leu Ala Trp Glu Ala Leu Cys Asp
100 105 110
Ala Gly Leu Pro Pro Thr Ser Leu Ala Gly Thr Asp Ala Ser Val Tyr
115 120 125
Val Ala Ala Asn Ser Asn Asp Tyr Gly Arg Arg Leu Leu Glu Asp Leu
130 135 140
Asp Arg Thr Gly Ala Trp Ala Val Asn Gly Thr Thr Phe Tyr Gly Ile
145 150 155 160
Ala Asn Arg Ile Ser Tyr Phe Leu Asp Ala His Gly Pro Ser Met Ala
165 170 175
Val Asp Thr Ala Cys Ala Gly Ser Leu Thr Ala Leu His Val Ala Gly
180 185 190
Gln Ala Leu His Arg Gly Glu Thr Ser Val Ala Ile Val Gly Gly Ile
195 200 205
Asn Ile Met Ala Ser Pro Ala Leu Val Val Ala Leu Asp Ala Ala Ser
210 215 220
Ala Thr Ser Pro Asp Gly Arg Ser Lys Ser Phe Asp Lys Ala Ala Asp
225 230 235 240
Gly Tyr Gly Arg Gly Glu Gly Gly Gly Val Val Val Leu Lys Arg Leu
245 250 255
Ser Asp Ala Val Arg Asp Gly Asp Pro Val His Gly Leu Val Leu Ala
260 265 270
Ser Gly Val Phe Gln Asp Gly Arg Ser Asp Gly Met Met Ala Pro Asn
275 280 285
Gly Ser Ala Gln Gln Arg Met Leu Glu Glu Ile Tyr Arg Arg Ser Gly
290 295 300
Ile Asp Pro Gly Thr Val Gln Tyr Val Glu Ala His Gly Thr Gly Thr
305 310 315 320
Gln Leu Gly Asp Ala Ala Glu Ala Gln Ala Ile Gly Asn Val Phe Gly
325 330 335
Pro Gly Arg Asp Gly Asp Asn Pro Leu Leu Ile Gly Thr Leu Lys Pro
340 345 350
Asn Val Gly His Val Glu Ala Ala Ser Gly Ile Ala Gly Val Ile Lys
355 360 365
Val Leu Leu Gly Met Arg His Gly Glu Leu Pro Pro Ser Pro His Glu
370 375 380
Glu Pro Asp Pro Gly Leu Gly Leu Glu Ala Arg Gly Leu Arg Leu Val
385 390 395 400
Ala Glu Pro Thr Pro Trp Pro Arg Gly Glu His Gly Met Arg Ala Gly
405 410 415
Val Ser Ser Tyr Gly Val Gly Gly Ser Ile Ala His Ala Val Leu Gln
420 425 430
Gln Ala Pro Pro Arg Pro Asp Arg Thr Glu Arg Pro Ala Ala Ala Ala
435 440 445
Thr Gly Arg Pro Gln Val Phe Pro Leu Ser Ala Ala Ser Glu Gln Gly
450 455 460
Val Arg Gly Leu Ala Gly Ser Val Ala Ala Trp Leu Arg Ala His Pro
465 470 475 480
Glu Thr Ala Leu Asp Asp Leu Ala His Thr Phe Thr Ala Arg Arg Ser
485 490 495
His Leu Ser Arg Arg Ala Ala Val Val Ala Gly Thr Thr Glu Glu Leu
500 505 510
Leu Gly Gly Leu Asp Ala Leu Ala Gly Gly Glu Lys Ser Pro Ala Val
515 520 525
Ala Leu Ala Ser Ala Ser Gly Phe Gly Asp Gly Gly Ala Ala Gly Pro
530 535 540
Ala Trp Val Phe Ser Gly His Gly Ala Gln Trp Ser Gly Met Gly Arg
545 550 555 560
Glu Leu Leu Thr Thr Glu Pro Val Phe Ala Gln Val Ile Asp Glu Leu
565 570 575
Ala Pro Val Phe Ser Glu Glu Leu Gly Trp Thr Pro Arg Glu Ala Ile
580 585 590
Glu Ala Gly Gly Pro Trp Thr Val Val Arg Thr Gln Ala Met Thr Phe
595 600 605
Ala Met Gln Val Ala Leu Ala Glu Val Trp Ser Asp Leu Gly Leu Arg
610 615 620
Pro Gly Ala Ile Ile Gly His Ser Val Gly Glu Ile Ala Ala Ala Ala
625 630 635 640
Val Ala Gly Ser Leu Asp Arg Ala Glu Ala Ala Arg Phe Ala Cys Arg
645 650 655
Arg Ala Arg Ala Leu Gly Lys Ile Ala Gly Arg Gly Ala Met Ala Met
660 665 670
Val Pro Met Ala Phe Ala Asp Val Glu Gln Arg Val Ala Gly Arg Asp
675 680 685
Ala Val Val Ala Ala Ile Ala Ala Ser Pro Leu Ser Thr Val Val Ser
690 695 700
Gly Asp Thr Ala Ala Val Glu Ala Leu Leu Ala Asp Leu Glu Ala Asp
705 710 715 720
Gly Ile Gln Ala Arg Arg Val Asn Thr Asp Val Ala Phe His Ser Pro
725 730 735
His Val Gln Glu Ile Leu Asp Glu Val Arg Gln Ala Ala Ala Ala Leu
740 745 750
Arg Ala Gly Thr Pro Arg Val Thr Leu Tyr Ser Thr Ala Leu Ala Asp
755 760 765
Pro Arg Ser Asp Ala Pro Arg Glu Gly Glu Tyr Trp Ala Thr Asn Leu
770 775 780
Ala Asp Pro Val Arg Phe His Gln Ala Val Arg Ala Ala Leu Asp Asp
785 790 795 800
Gly Thr Arg Val Phe Leu Glu Val Ser Ser His Pro Val Val Ala His
805 8l0 815
Ser Ile Thr Glu Thr Ala Leu Asp Ala Gly Val Pro Asp Ala His Val
820 825 830
Ala Ile Thr Leu Arg Arg Glu Gln Pro Glu Gln Arg Thr Val Leu Ala
835 840 845
Asn Leu Ala Arg Leu His Ser Leu Gly Thr Pro Val Thr Trp Ser Tyr
850 855 860
Asp Gly Asp Leu Val Asp Val Pro Ala Val Arg Trp Gln His Lys Pro
865 870 875 880
Tyr Trp Ile Phe Pro Asp Thr Ala Pro Glu Gln Gly Ala Gly Leu Gly
885 890 895
His Asp Pro Gln Thr His Thr Leu Ile Gly Ala Arg Thr Thr Val Ala
900 905 910
Ser Ala Pro Val Gln Arg Val Trp Gln Thr Glu Leu His Met Glu Asn
915 920 925
Arg Pro Tyr Ala Gln Ser His Lys Val Val Gly Val Glu Thr Val Pro
930 935 940
Ala Ser Val Val Leu Asn Ser Phe Ile Thr Ala Ala Thr Asn Glu Gly
945 950 955 960
Glu Arg Ala Cys Gly Leu Arg Asp Ile Val Phe Arg Ile Pro Leu Ala
965 970 975
Ala His Pro Thr Arg Val Val Gln Val Val Leu Glu Gln Asp Lys Val
980 985 990
Arg Ile Ala Ser Arg Ile Lys Arg Asp Gln Glu Ser Gly Gly Val Arg
995 1000 1005
Asp Asp Glu Trp Leu Thr His Thr Thr Ala Thr Val Val His Glu Pro
1010 1015 1020
Glu Val Gly Ala Arg Pro Met Glu Asp Pro Asp Val Ile Arg Ala Arg
1025 1030 1035 1040
Cys Pro Val Ser Trp Thr Trp Ala Lys Val Asp Gly Ile Phe Arg Thr
1045 1050 1055
Met Gly Val Asp Gly Tyr Thr Phe Pro Trp Val Val Glu Glu Leu Leu
1060 1065 1070
Arg Gly Glu Asp Glu Gln Phe Ser Thr Ile Thr Val Asp His Thr Pro
1075 1080 1085
Lys Leu His Pro Ser Ser Trp Thr Ala Val Val Asp Ala Ala Leu Thr
1090 1095 1100
Ala Ser Gly Val Leu Val Met Asp Glu Asn Ser Asn Val Leu Arg Thr
1105 1110 1115 1120
Cys Ser His Leu Glu Ser Leu Ser Phe Val Gly Pro Pro Pro Pro Arg
1125 1130 1135
Ile His Val His Thr Val Arg Asp Pro Arg Thr Pro Asp Thr Ile Ser
1140 1145 1150
Met Thr Val Ala Asp Glu Ser Gly Ala Val Val Cys Glu Ala Arg Gly
1155 1160 1165
Leu Arg Tyr Val Lys Val Gln Asp Ile Gly Ser Gly Ala Val Gly Pro
1170 1175 1180
Arg Asp Leu Val His Glu Leu Ala Trp Glu Pro Val Glu Val Pro Ala
1185 1190 1195 1200
Asp Ala Pro Val Pro Ser Gln Ala Leu Val Val Gly Gly Ala Ala Gly
1205 1210 1215
Gly Pro Ala Leu Val Glu Ala Leu Thr Ala Arg Gly Val Arg Ala Arg
1220 1225 1230
Ala Val Pro Asp Ala Thr Ala Ile Gly Asp Ala Ser Leu Thr Cys Ala
1235 1240 1245
Asp Val Val Val Val Ala Pro Glu Ala Leu Leu Pro Gly Glu Ala Pro
1250 1255 1260
Glu Gln Ala Ala Arg Arg Cys Ala Gln Leu Leu Val Asp Ala Val Gln
1265 1270 1275 1280
Gln Val Ala Ala Val Pro Asp Glu Arg Arg Arg Pro Arg Val Trp Ala
1285 1290 1295
Leu Thr Arg Glu Val Arg Ala Gly Ala Thr Glu Ala Ala Leu Ala His
1300 1305 1310
Ala Pro Leu Trp Gly Ala Gly Arg Ile Val Ala Gly Glu Arg Pro Asp
1315 1320 1325
Leu Trp Gly Gly Val Ile Asp Val Ala Glu Asn Ala Val Pro Gln Gln
1330 1335 1340
Val Ala Ser Leu Ile Gly Ala Leu Pro His Thr Glu Asp Val Leu Ser
1345 1350 1355 1360
Leu Asp Ser Glu Gly Val Thr Ala Ala Arg Leu Arg Gln Val Ala Arg
1365 1370 1375
Pro Ala Glu Arg Glu Pro Val Asp Cys Arg Pro Asp Gly Thr Tyr Leu
1380 1385 1390
Val Thr Gly Gly Leu Gly Ala Leu Gly Leu Glu Ala Ala Arg His Leu
1395 1400 1405
Val Glu Gln Gly Ala Arg Arg Leu Val Leu Ile Gly Arg Arg Gly Leu
1410 1415 1420
Pro Ser Arg Ser Arg Trp Asp Gln Val Asp Asp Pro Ala Val Ala Ala
1425 1430 1435 1440
Gln Ile Ala Glu Val Val Ala Leu Glu Ala Ala Gly Ala Thr Val Arg
1445 1450 1455
Val Leu Ser Leu Asp Ile Ser Asp Ala Glu Ala Thr Ala Arg Ala Leu
1460 1465 1470
Asp Pro Gly Ala Leu Asp Met Pro Pro Val Arg Gly Ile Val His Cys
1475 1480 1485
Ala Gly Val Val Ser Asp Ala Leu Val Glu Lys Thr Gly Ala Ala Asn
1490 1495 1500
Leu Asp Thr Thr Met Gly Pro Lys Ala Asp Gly Ala Met Val Leu His
1505 1510 1515 1520
Arg Leu Phe Pro Ala Gly Thr Leu Asp Phe Phe Thr Met Phe Ser Ser
1525 1530 1535
Cys Gly Gln Leu Ala Arg Leu Thr Gly Gln Val Ser Tyr Ala Ser Ala
1540 1545 1550
Asn Ser Phe Leu Asp Ala Leu Ala Ala Leu Arg Arg Ser Arg Gly Glu
1555 1560 1565
Thr Gly Thr Thr Ser Phe Ala Trp Ala Gln Trp Ile Gly Arg Gly Met
1570 1575 1580
Gly Glu Thr Thr Gly Arg Ala Thr Ile Leu Glu Ala Glu Ser Arg Gly
1585 1590 1595 1600
Leu Gly Gly Ile Thr Val Ser Glu Ala Leu Arg Ser Trp Ala Tyr Ala
1605 1610 1615
Asp Arg Phe Ala Leu Pro Tyr Ala Ala Val Met Arg Val Met Pro Asp
1620 1625 1630
His Thr Leu Pro Val Phe Ser His Leu Ser Val Thr Asp Ala Gly Ala
1635 1640 1645
Gln Ser Ala Asp Ala Gly Gly Val Asp Trp Ala Thr Val Pro Ala Gly
1650 1655 1660
Glu Leu Pro Glu Leu Val Leu Lys Val Thr His Glu Gln Val Ala Ala
1665 1670 1675 1680
Glu Leu Asn Leu Ala Val Asp Asp Ile Ala Ile Asp Gln Pro Leu Leu
1685 1690 1695
Glu Leu Gly Val Asp Ser Val Leu Thr Val Ala Leu Arg Val Arg Leu
1700 1705 1710
His Arg Cys Phe Ala Val Asp Leu Pro Pro Thr Ile Leu Trp Ser Asn
1715 1720 1725
Pro Thr Val Arg Ala Leu Ala Glu Phe Leu Ala Ala Glu Val Gly Gly
1730 1735 1740
Ala Thr Ala Asp Ala Glu Glu Thr Asp Pro Val Ala Gly Leu Pro Ala
1745 1750 1755 1760
Pro Gln Gln Gly Ser Gly Thr Ala Glu Gln Leu Asp Ala Val Ala Ala
1765 1770 1775
Ala Ala Gly
<210>27
<211>257
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Pro Gly Gln Asp Ser Val Leu Ile Glu Glu Ser Ile Pro Gly Arg
1 5 10 15
Leu Arg Ala Asp His Phe Ala Gln His Gly Trp Ala His Gly Arg Leu
20 25 30
Ser Val Pro Thr Glu Gly Ser Asp Trp Glu Gln Phe Ala Ser Ala Trp
35 40 45
Asp Glu Leu Val Pro Asp Pro Tyr Leu Gly Glu Glu Arg Cys Arg Arg
50 55 60
Asn Arg Arg Phe Gly Arg Val Leu Ala His Arg Asp Gly Ala Leu Glu
65 70 75 80
Pro Leu Arg Gly Ser Asp Phe Phe Gln Ser Lys Glu Ile Asn Arg Ala
85 90 95
Phe Gly Gly Arg Leu Arg Val Phe Glu Pro Leu Thr Asp Ala Val Leu
100 105 110
Thr Gly Pro Cys Leu Pro Arg Leu Leu Arg Glu Thr Val Ala Val Val
115 120 125
Asn Glu Ala Ala Gly Lys Gln Asp Trp Glu Leu Gly Ile His Phe Ile
130 135 140
Arg Val Ile Ala Asp Pro Gly Glu Ser Ser Glu Pro Ala Pro Glu Gly
145 150 155 160
Arg His Ser Asp Gly His Ala Tyr Val Ala Ile His Leu Ile Asp Arg
165 170 175
His Gln Cys Ala Gly Gly Arg Asn Gln Leu Phe Arg Asn Asn Glu Ser
180 185 190
Arg Ala Gln Phe Ser Val Val Met Thr Glu Pro Leu Glu Thr Leu Ile
195 200 205
Val Ser Asp Thr Thr Met Glu His Ser Val Ser Glu Ile Arg Pro Glu
210 215 220
Ala Gly Thr Gly Ser Gly Trp Arg Asp Thr Met Ile Val Asp Phe Asn
225 230 235 240
Arg Val Pro Val Pro Glu Glu Val Thr Gly Arg Thr Tyr Gly Phe Ile
245 250 255
Arg
<210>28
<211>221
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Thr His Val Ala Glu Thr Ser Ala Pro Thr Arg Ser Glu Pro Asp
1 5 10 15
Thr Arg Val Leu Thr Leu Pro Gly Thr Ala Ser Ala Pro Glu Phe Arg
20 25 30
Leu Ile Asp Ile Asp Gly Leu Leu Asn Asn Arg Ala Thr Thr Asp Val
35 40 45
Arg Asp Leu Gly Ser Gly Arg Leu Asn Ala Trp Gly Asn Ser Phe Pro
50 55 60
Ala Ala Glu Leu Pro Ala Pro Gly Ser Leu Ile Thr Val Ala Gly Ile
65 70 75 80
Pro Phe Thr Trp Ala Asn Ala His Ala Arg Gly Asp Asn Ile Arg Cys
85 90 95
Glu Gly Gln Val Val Asp Ile Pro Pro Gly Gln Tyr Asp Trp Ile Tyr
100 105 110
Leu Leu Ala Ala Ser Glu Arg Arg Ser Glu Asp Thr Ile Trp Ala His
115 120 125
Tyr Asp Asp Gly His Ala Asp Pro Leu Arg Val Gly Ile Ser Asp Phe
130 135 140
Leu Asp Gly Thr Pro Ala Phe Gly Glu Leu Ser Ala Phe Arg Thr Ser
145 150 155 160
Arg Met His Tyr Pro His His Val Gln Glu Gly Leu Pro Thr Thr Met
165 170 175
Trp Leu Thr Arg Val Gly Met Pro Arg His Gly Val Ala Arg Ser Leu
180 185 190
Arg Leu Pro Arg Ser Val Ala Met His Val Phe Ala Leu Thr Leu Arg
195 200 205
Thr Ala Ala Ala Val Arg Leu Ala Glu Gly Ala Thr Thr
210 215 220
<210>29
<211>337
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Thr Thr Thr Ala Pro Pro Val Glu Leu Trp Thr Arg Asp Leu Gly
1 5 10 15
Ser Cys Leu His Gly Thr Leu Ala Thr Ala Leu Ile Arg Asp Gly His
20 25 30
Asp Pro Val Thr Val Leu Gly Ala Pro Trp Glu Phe Arg Arg Arg Pro
35 40 45
Gly Ala Trp Ser Ser Glu Glu Tyr Phe Phe Phe Ala Glu Pro Asp Ser
50 55 60
Leu Ala Gly Arg Leu Ala Leu Tyr His Pro Phe Glu Ser Thr Trp His
65 70 75 80
Arg Ser Asp Gly Asp Gly Val Asp Asp Leu Arg Glu Ala Leu Ala Ala
85 90 95
Gly Val Leu Pro Ile Ala Ala Val Asp Asn Phe His Leu Pro Phe Arg
100 105 110
Pro Ala Phe His Asp Val His Ala Ala His Leu Leu Val Val Tyr Arg
115 120 125
Ile Thr Glu Thr Glu Val Tyr Val Ser Asp Ala Gln Pro Pro Ala Phe
130 135 140
Gln Gly Ala Ile Pro Leu Ala Asp Phe Leu Ala Ser Trp Gly Ser Leu
145 150 155 160
Asn Pro Pro Asp Asp Ala Asp Val Phe Phe Ser Ala Ser Pro Ser Gly
165 170 175
Arg Arg Trp Leu Arg Thr Arg Met Thr Gly Pro Val Pro Glu Pro Asp
180 185 190
Arg His Trp Val Gly Arg Val Ile Arg Glu Asn Val Ala Arg Tyr Arg
195 200 205
Gln Glu Pro Pro Ala Asp Thr Gln Thr Gly Leu Pro Gly Leu Arg Arg
210 215 220
Tyr Leu Asp Glu Leu Cys Ala Leu Thr Pro Gly Thr Asn Ala Ala Ser
225 230 235 240
Glu Ala Leu Ser Glu Leu Tyr Val Ile Ser Trp Asn Ile Gln Ala Gln
245 250 255
Ser Gly Leu His Ala Glu Phe Leu Arg Ala His Ser Val Lys Trp Arg
260 265 270
Ile Pro Glu Leu Ala Glu Ala Ala Ala Gly Val Asp Ala Val Ala His
275 280 285
Gly Trp Thr Gly Val Arg Met Thr Gly Ala His Ser Arg Val Trp Gln
290 295 300
Arg His Arg Pro Ala Glu Leu Arg Gly His Ala Thr Ala Leu Val Arg
305 310 315 320
Arg Leu Glu Ala Ala Leu Asp Leu Leu Glu Leu Ala Ala Asp Ala Val
325 330 335
Ser
<210>30
<211>287
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Pro Ile Ala Val Val Ala Ser Arg Ile Arg Arg Glu Glu Arg Leu
1 5 10 15
Ile Leu Glu Ala Phe Glu Arg Leu Gly Leu Arg Cys Glu His Val Asp
20 25 30
Pro Arg Gly Leu Arg Leu Leu Leu Ala Asp Gly Arg Pro Pro Tyr Arg
35 40 45
Ala Ala Leu Met Arg Glu Val Ser His Ser Arg Ala Ala Ser Val Ala
50 55 60
Thr Thr Leu Glu Ala Leu Gly Val Pro Thr Tyr Asn Arg Pro Arg Val
65 70 75 80
Leu Asp Val Cys Gly Asp Lys Leu Arg Thr Ala Leu Ala Phe His Ser
85 90 95
Ala Gly Leu Arg Met Pro Gln Ala Ala Ala Thr Trp Gly Thr Gly Ala
100 105 110
Ala Leu Glu Ala Met Pro Lys Leu Gly Tyr Pro Val Val Val Lys Pro
115 120 125
Val Thr Gly Ser Trp Gly His Leu Thr Thr Arg Val Arg Asp Glu Glu
130 135 140
Gln Gly Arg Ala Val Leu Glu His Arg Ala Ala Leu Pro Asn Pro Gln
145 150 155 160
Gln His Val Phe Phe Leu Gln Glu His Ile Asp Lys Pro Gly Arg Asp
165 170 175
Ile Lys Ala Tyr Val Ala Gly Asp Arg Val Ile Cys Ala Ile Tyr Lys
180 185 190
Asn Ala Val Asp Asp Trp Arg Thr Asn Thr Ala Ile Gly Gly Ala Ala
195 200 205
Thr Pro Cys Pro Ile Thr Pro Glu Leu Thr Glu Leu Ala Val Ala Ala
210 215 220
Ala His Ala Val Gly Gly Gly Phe Leu Gly Val Asp Leu Leu Ile Asp
225 230 235 240
Gln Gln Glu Arg Leu Tyr Ala Asn Glu Val Asn His Thr Pro Glu Phe
245 250 255
His Gly Ala Val Asp Ala Thr Gly Ile Asp Val Ala Gln Gly Ile Val
260 265 270
Glu Trp Ala Val Ala Glu Ile Glu Ala Gln Glu Arg Ala Ala Ala
275 280 285
<210>31
<211>243
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ser Ala Thr Ala Gly Pro Pro Ser Leu Leu Ala Leu Trp Ser Ala
1 5 10 15
Pro Arg Ser Arg Ser Thr Val Phe Phe Arg Met Met Val Glu Arg Gln
20 25 30
Asp Val Leu Ala Leu His Glu Pro Phe Cys Asn Ile Ala Asn Asp Gly
35 40 45
Ser Thr Val Ile Asp Gly Arg Val Val His Ser Thr Asp Thr Leu Thr
50 55 60
Asp Thr Leu Ile Gly Leu Ala Ala Gln Arg Thr Val Phe Phe Lys Asp
65 70 75 80
Thr Thr Asp Cys Phe Tyr Asp Ser Val Phe Ala Arg Pro Asp Phe Leu
85 90 95
Lys Ser Gly His His Ala Phe Leu Leu Arg Asn Pro Lys Glu Ile Ile
100 105 110
Pro Ser Tyr Ala Ala Ile Lys Pro Asp Met Ala Cys His Glu Val Gly
115 120 125
Ile Ala Tyr Leu His Arg Ile His Gln Ala Val Leu Asp Ala Gly Gly
130 135 140
Ser Ala Val Ile Leu Asp Ser Asp Asp Phe Val Asp His Pro Glu Ala
145 150 155 160
Thr Val Arg Gln Tyr Cys Ala Thr Val Gly Leu Pro Phe Arg Ala Asp
165 170 175
Ala Leu Gln Trp Gln Pro Gly Gln Arg Pro Glu Trp Ser Gln Ser Ala
180 185 190
Arg Trp His Thr Asp Val Ser Ala Thr Ser Thr Ile Arg Arg Arg Glu
195 200 205
Arg Thr Tyr Asp Arg Thr Thr Asp Asn Asp Pro Leu Leu Gln Arg Phe
210 215 220
Tyr Glu His His Ile Pro Phe Tyr Glu Tyr Leu Trp Glu Arg Arg Leu
225 230 235 240
Arg Ile Ser
<210>32
<211>419
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Ile Ile Ser Lys Pro Pro Phe Cys Pro Val Gln Phe Gln Asp Ser
1 5 10 15
Asp Leu Leu Asn Ser Gly Phe His Ala Ala Thr Asp Met His Ala Leu
20 25 30
Trp Ala Asp Leu Arg Glu Asn Gln Pro Leu Tyr Arg Ala Asp Pro Gly
35 40 45
Gly Gly Arg Glu Pro Phe Trp Val Val Thr Arg His Glu Asp Val Ser
50 55 60
Arg Val Leu Arg Ser His Gly Glu Phe Ser Ser Arg Arg Gly Thr Ile
65 70 75 80
Leu Cys Val Leu Asp Leu Asn Thr Ala Asp Ile Ala Ser Asp Asn Met
85 90 95
Met Ala Asp Thr Asp Pro Pro Arg His Arg Glu Phe Arg Glu Pro Leu
100 105 110
Asn Lys Ala Phe Ala Pro Ser Val Val Ala Thr Gln Glu Arg Leu Leu
115 120 125
Arg Glu Leu Ser Gln Asp Leu Ile Gln Ser Val Leu Glu Ala Gly Val
130 135 140
Tyr Asp Ile Ala His Lys Thr Met Met Phe Pro Met Ala Val Thr Gly
145 150 155 160
Thr Leu Met Gly Leu Pro Pro Glu Ser Trp Glu Arg Leu Ala Glu Leu
165 170 175
Val Met Met Thr Ile Ala Tyr Asp Asp Pro Asp Tyr Ser Ala Gly Ser
180 185 190
Ala Gln Ala Thr Val Arg Gln Ala Arg His Glu Leu Phe Glu Tyr Phe
195 200 205
Gln Lys Glu Tyr Ala Gln Arg Ser Arg Asp Asp Val Asp Pro Asp Val
210 215 220
Ile Gly Ala Met Val Gly Met Asp Leu Ser Glu Gly Pro Met Thr Gln
225 230 235 240
Glu Gln Val Leu Leu Asn Ala Phe Val Leu Leu Ile Gly Ala Asn Val
245 250 255
Thr Thr Pro His Ala Leu Cys Thr Leu Met Ser Val Met Ala Glu His
260 265 270
Pro Glu Gln Tyr Arg Ala Val Gln Glu Asn Pro Glu Leu Arg Ala Ser
275 280 285
Cys Leu Gln Glu Leu Leu Arg Trp Ser Ser Pro Val Thr Ala Leu Met
290 295 300
Arg Tyr Ala Val Lys Asp Val Glu Met His Gly Arg Thr Ile Arg Ala
305 310 315 320
Gly Glu Pro Val Thr Ala Trp Ile Gly Ala Ala Asn Arg Asp Glu Arg
325 330 335
Val Phe Pro Asp Pro Tyr Thr Phe Asp Val Ala Arg Arg Pro Asn Lys
340 345 350
His Leu Ala Phe Gly Leu Gly Pro His Tyr Cys Ile Gly Ala Asn Leu
355 360 365
Ala Lys Val Gly Leu Asp Ile Phe Leu Asp Glu Leu Leu Glu Arg Val
370 375 380
Glu Ser Ile Glu Ile Ala Gly Glu Val Lys His Val Ala Ser His Phe
385 390 395 400
Val Pro Gly Tyr Lys Ser Met Pro Val Arg Phe Thr Pro Arg Glu Ala
405 410 415
His Val Arg
<210>33
<211>249
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val His Arg Ser Asp Ala Arg Leu Trp Phe Asp Gly Arg Phe Arg Ser
1 5 10 15
Ala Thr Ala Thr His Ser Leu Tyr Cys Leu Pro Phe Ala Gly Gly Ser
20 25 30
Ala Thr Tyr Tyr Ala Asp Trp Ala Pro His Cys Ala Ser Pro Val Glu
35 40 45
Leu Val Pro Val Gln Leu Pro Gly Arg Gly Gly Arg Met Thr Glu Ser
50 55 60
Ser Ala Lys Asp Leu Val Gln Leu Ala Glu Glu Ile Ala Asp Thr Ile
65 70 75 80
Ala Ala Glu Pro Thr Arg Thr Leu Leu Tyr Gly His Ser Met Gly Ala
85 90 95
Met Leu Ala Phe Glu Val Ser Arg Arg Leu Gln Thr Leu Asn Arg Pro
100 105 110
Val Arg His Leu Phe Val Ser Gly Arg Pro Ala Pro Thr Ile Val Arg
115 120 125
Pro Ile Ala Pro Val Ser His Leu Pro Arg Ala Glu Phe Ile Gln Met
130 135 140
Leu Arg Asp Tyr Gly Ala Ala Asp Gln Thr Val Phe Glu His Asp Glu
145 150 155 160
Leu Leu Asp Leu Leu Met Pro Met Ile Arg Ala Asp Phe Ala Met Ile
165 170 175
Glu Asn Tyr Arg Tyr Gln Glu Ala Pro Arg Leu Ser Cys Pro Ile Ser
180 185 190
Ala Trp Cys Gly Asp Ala Asp Pro Glu Val Pro Pro Thr Ala Met Arg
195 200 205
Gly Trp Gly Asp Gln Thr Ser Gly Glu Phe Thr Leu Ser Val Leu Arg
210 215 220
Gly Gly His Phe Phe Leu Thr Glu His Arg Ala Glu Ile Met Arg Ala
225 230 235 240
Val Leu Ala Ala Val Arg Arg Ala Arg
245
<210>34
<211>382
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Val Pro Pro Gln Ser Val Asp Phe Ala Asn Glu Val Glu Glu Ile Leu
1 5 10 15
Thr Ala Pro Glu Thr Glu Arg Leu Leu Arg Glu Leu Ala Thr Arg Gln
20 25 30
Asp Gly Met Asp Gly Asp Pro Arg Ala Leu Tyr Arg His Leu Gly Asp
35 40 45
Ala Gly Ile Leu Ala Pro Ser Trp Pro Val Glu Tyr Gly Gly Arg Gly
50 55 60
Ala Asp Phe Thr Ala Thr Val Ala Leu Leu Glu Lys Leu Val Ala His
65 70 75 80
Arg Ile Pro Gln Asn Leu Tyr Cys Ile Ser Val Gln Asn Val Gly Ser
85 90 95
Leu Ile Leu Ala Ala Gly Ser Glu Gln Gln Arg Arg Thr Leu Leu Pro
100 105 110
Ala Leu Ala Ser Thr Glu Leu Thr Ser Cys Ile Leu Phe Thr Glu Pro
115 120 125
Gly Asn Gly Ser Asp Leu Ala Gly Ile Thr Thr Thr Ala Val Arg Asp
130 135 140
Gly Thr Gly Trp Val Leu Asn Gly Arg Lys Thr Tyr Asn Leu Lys Ser
145 150 155 160
Ala Tyr Ala Asp Ile Ala Leu Ile Ala Val Arg Thr Asp Pro Gln Ser
165 170 175
Ser Gln Tyr Glu Gly Leu Thr Leu Phe Leu Val Pro Leu Asp Ala Pro
180 185 190
Gly Val Val Ile Arg Pro Ile Pro Ser Leu Ala Asp Glu Gln Phe His
195 200 205
Asp Ile Trp Phe Thr Asp Val Arg Val Asp Glu Ser Ala Leu Phe Gly
210 215 220
Gln Val Gly Glu Gly Trp Ser Leu Ile Thr Gln Met Phe Ala Ala Glu
225 230 235 240
Arg Thr Gly Leu Asp Tyr Tyr Ala Arg Gly Arg His Trp Leu Asp Leu
245 250 255
Val Ala Gln Arg Val Asp Arg Asn Gly Ala Arg Gln Ser Asp Ala Val
260 265 270
Arg Thr Glu Leu Ala Arg His Glu Ala Arg Ile Thr Ala Ser Lys Leu
275 280 285
Leu Ser Cys Gln Val Met Gln Asn Leu Gln Asp Gly Ser Pro Asp Ile
290 295 300
Ala Glu Ser Ser Phe Ala Lys Trp His Cys Ser Glu Ser Ala Gln Arg
305 310 315 320
Ile Ala Trp Trp Ala Leu Asp Thr Leu Gly Gln Asp Leu Leu Glu Pro
325 330 335
Gly Pro Asp Pro Gly Asp Arg Thr Leu Glu Ala Ala Phe Arg G1u Ala
340 345 350
Pro Gly Met Thr Ile Ser Gly Gly Ala Ser Glu Val Met Leu Asp Ile
355 360 365
Leu Ser Ser Ala Arg Val Phe Thr His Ala Pro Arg Glu Asp
370 375 380
<210>35
<211>333
<212>PRT
<213>Streptomyces sahachiroi NRRL2485
<400>1
Met Asp Leu Arg Pro Asp Pro Phe Gln Gln Arg Leu Arg Ser Ala Val
1 5 10 15
Asp Thr Val Leu Thr Arg Thr Gly Asp Leu Pro Ala Gln Leu Ala Asp
20 25 30
Ile Gly Val Pro Ser Leu Gly Ala Pro Glu Arg Leu Gly Gly Phe Ala
35 40 45
Leu Gly Leu Ser Ala Asp Ile Val Val Asn Glu Arg Leu Gly His Gly
50 55 60
Leu Glu Pro Leu Asp Ser Tyr Arg Ala Thr Ala Phe Ala Leu Asp Leu
65 70 75 80
Leu Asp Cys Asp Gly Ala Pro Thr Glu Leu Leu Ala Glu Ile Leu Ala
85 90 95
Asp Leu Tyr Lys Gly Thr Arg His Ala Val Val Val Gly Ala Arg Gly
100 105 110
Gly Gly Ala Val Gln Val Arg Asp Gly Gly Arg Leu Trp Gly Glu Ser
115 120 125
Glu Ser Leu Pro Ala Gly Asp Ile Ala Leu Cys Ile Val Arg Thr Ala
130 135 140
Gly Glu Asp Gly Ala Ala Trp Arg Leu Ala Ile Pro Glu Pro Gly Thr
145 150 155 160
Cys Val Thr Glu Pro Ala Glu His Leu Gly Ile Pro Ala Ser Arg Leu
165 170 175
Arg Phe His Gly Ala Pro Ser Gln Ser Leu Pro Val Thr Thr Ala His
180 185 190
Trp Gln Arg Ala Leu Ala Ala Ala Arg Ile Arg Gln Ala Ala Leu Leu
195 200 205
Leu Gly Ile Ala Asp Trp Ile Thr Asp Val Ala Arg Ala His Val Asn
210 215 220
Thr Arg Thr Gln Ser Gly Lys Pro Leu Val Glu Arg Gln Thr Val Ala
225 230 235 240
His Arg Leu Ala Val Leu Ile Gly Glu Gly Glu Gly Trp Lys Leu Leu
245 250 255
Leu Asn Lys Ala Ala Trp Glu Phe Asp Arg Gly Gly Arg Pro Asp Ala
260 265 270
Ala Ala Met Leu Ala Val Ala Gly Glu His Ala Gln Leu Ala Ser Arg
275 280 285
Ile Ala Leu Gln Leu His Gly Val Arg Gly Met Leu Ala His Ser Thr
290 295 300
Ala Ala Ser Val Tyr Arg Met Val Ala Val Glu Ser Ala Arg Leu Gly
305 310 315 320
Thr Pro Ala Ala Leu Trp Arg Glu Ala Gly Ser Ala Ala
325 330
Claims (9)
1. 一种抗肿瘤活性的抗生素-阿嗪霉素Azinomycin的生物合成基因簇,其特征在于编码阿嗪霉素生物合成所涉及的34个基因,具体为:
1)1个重复使用的I型聚酮合成酶基因,即aziB:
aziB位于基因簇核苷酸序列第38515-43854个碱基处,长度为5340个碱基对,编码I型聚酮合成酶,1779个氨基酸;
2)萘环修饰酶基因,即aziB1,aziB2共个基因:
aziB1位于基因簇核苷酸序列第12587-13792个碱基处,长度为1206个碱基对,编码细胞色素P450氧化酶,401个氨基酸;
aziB2位于基因簇核苷酸序列第11548-12585个碱基处,长度为1038个碱基对,编码O-甲基转移酶,345个氨基酸;
3)非核糖体聚肽骨架合成及修饰酶基因,即aziA1,aziA2,aziA3,aziA4,aziA5,aziA6,aziA7,aziA8共8个基因:
aziA1位于基因簇核苷酸序列第13873-15756个碱基处,长度为1884个碱基对,编码非核糖体聚肽合成酶,627个氨基酸;
aziA2位于基因簇核苷酸序列第17541-20888个碱基处,长度为3348个碱基对,编码非核糖体聚肽合成酶,1115个氨基酸;
aziA3位于基因簇核苷酸序列第2596-6387个碱基处,长度为3792个碱基对,编码非核糖体聚肽合成酶,1263个氨基酸;
aziA4位于基因簇核苷酸序列第7202-10513个碱基处,长度为3312个碱基对,编码非核糖体聚肽合成酶,1103个氨基酸;
aziA5位于基因簇核苷酸序列第33636-38261个碱基处,长度为4626个碱基对,编码非核糖体聚肽合成酶,1541个氨基酸;
aziA6位于基因簇核苷酸序列第6384-7205个碱基处,长度为822个碱基对,编码硫酯酶,273个氨基酸;
aziA7位于基因簇核苷酸序列第10796-11548个碱基处,长度为753个碱基对,编码硫酯酶,250个氨基酸;
aziA8位于基因簇核苷酸序列第49525-50274个碱基处,长度为750个碱基对,编码硫酯酶,249个氨基酸;
4)非天然氨基酸结构单元合成酶基因,即aziC1,aziC2,aziC3,aziC4,aziC5,aziC6,aziC7,aziC8,aziC9,aziC10,aziC11共11个基因:
aziC1位于基因簇核苷酸序列第个47453-48484碱基处,长度为732个碱基对,编码分枝氨基酸氨基转移酶,243个氨基酸;
aziC2位于基因簇核苷酸序列第46593-47456个碱基处,长度为864个碱基对,编码赖氨酸生物合成酶,287个氨基酸;
aziC3位于基因簇核苷酸序列第30737-31615个碱基处,长度为879个碱基对,编码N-己酰谷氨酸激酶,292个氨基酸;
aziC4位于基因簇核苷酸序列第29735-30658个碱基处,长度为924个碱基对,编码N-己酰-γ-谷氨酰磷酸还原酶,307个氨基酸;
aziC5位于基因簇核苷酸序列第27040-27975个碱基处,长度为936个碱基对,编码酮基转移酶N-端亚单位,311个氨基酸;
aziC6位于基因簇核苷酸序列第26021-27043个碱基处,长度为1023个碱基对,编码酮基转移酶C-端亚单位,340个氨基酸;
aziC7位于基因簇核苷酸序列第27972-29294个碱基处,长度为1323个碱基对,编码氨基转移酶,440个氨基酸;
aziC8位于基因簇核苷酸序列第32741-33559个碱基处,长度为819个碱基对,编码己酰-CoA脱氢酶,272个氨基酸;
aziC9位于基因簇核苷酸序列第48254-49513个碱基处,长度为1260个碱基对,编码细胞色素P450氧化酶,419个氨基酸;
aziC10位于基因簇核苷酸序列第31608-32738个碱基处,长度为1131个碱基对,编码N-己酰鸟氨酸/N-己酰赖氨酸脱酰基酶,376个氨基酸;
aziC11位于基因簇核苷酸序列第25755-26018个碱基处,长度为264个碱基对,编码肽酰载体蛋白,87个氨基酸;
5)抗性基因,即aziE:
aziE位于基因簇核苷酸序列第15839-17239个碱基处,长度为1401个碱基对,编码转运蛋白,466个氨基酸;
6)后修饰酶基因,即aziD1,aziD2,aziD3共3个基因:
aziD1位于基因簇核苷酸序列第24547-25758个碱基处,长度为1212个碱基对,编码O-酰基转移酶,403个氨基酸;
aziD2位于基因簇核苷酸序列第51462-52463个碱基处,长度为1002个碱基对,编码己酰-CoA脱氢酶,333个氨基酸;
aziD3位于基因簇核苷酸序列第50308-51456个碱基处,长度为1149个碱基对,编码己酰-CoA脱氢酶,382个氨基酸;
7)还包括8个功能不明确的基因,即aziH1,aziH2,aziH3,aziF,aziG,aziU1,aziU2,aziU3:
aziH1位于基因簇核苷酸序列第22185-22997个碱基处,长度为813个碱基对,编码硫酸腺苷酰转移酶亚单位2,270个氨基酸;
aziH2位于基因簇核苷酸序列第20878-22185个碱基处,长度为1308个碱基对,编码硫酸腺苷酰转移酶亚单位1,435个氨基酸;
aziH3位于基因簇核苷酸序列第23101-23658个碱基处,长度为558个碱基对,编码腺苷酰硫酸激酶,185个氨基酸;
aziF位于基因簇核苷酸序列第24328-24516个碱基处,长度为189个碱基对,编码MbtH类似蛋白,62个氨基酸;
aziG位于基因簇核苷酸序列第23726-24127个碱基处,长度为402个碱基对,编码硫酯酶,133个氨基酸;
aziU1位于基因簇核苷酸序列第44054-44827个碱基处,长度为774个碱基对,编码未知功能蛋白,257个氨基酸;
aziU2位于基因簇核苷酸序列第44838-45503个碱基处,长度为666个碱基对,编码未知功能蛋白,221个氨基酸;
aziU3位于基因簇核苷酸序列第45500-46513个碱基处,长度为1014个碱基对,编码未知功能蛋白,337个氨基酸。
2. 根据权利要求1所述的阿嗪霉素的生物合成基因簇,其特征在于编码的非核糖体聚肽合成酶包含下述模块或结构域:酰基辅酶A连接酶结构域AL、肽酰缩合酶结构域C、腺苷化酶结构域A、肽酰基载体蛋白PCP、端基还原酶结构域RE。
3. 根据权利要求1所述的阿嗪霉素的生物合成基因簇,其特征在于编码的聚酮合成酶包含下述模块或结构域:酮基合成酶结构域KS、酰基转移酶结构域AT、酮基还原酶结构域KR、脱水酶结构域DH、酰基载体蛋白ACP。
4. 根据权利要求1所述的阿嗪霉素的生物合成基因簇的编码蛋白用于催化合成抗生素阿嗪菌素及其类似物。
5. 根据权利要求4所述的阿嗪霉素的生物合成基因簇的编码蛋白用于催化合成3-甲氧基-5-甲基萘甲酸。
6. 根据权利要求4所述的阿嗪霉素的生物合成基因簇的编码蛋白用于催化合成非天然氨基酸三肽骨架。
7. 根据权利要求4所述的阿嗪霉素的生物合成基因簇的用途,对其中的基因进行遗传改造获得的突变体经生物发酵产生3-甲氧基-5-甲基萘甲酸。
8. 根据权利要求4所述的阿嗪霉素的生物合成基因簇的用途,其中的基因在Streptomyces albus J1074中进行异源表达产生5-甲基萘甲酸。
9. 根据权利要求4所述的阿嗪霉素的生物合成基因簇的用途,其中的基因在Streptomyces albus J1074中进行异源表达产生3-甲氧基-5-甲基萘甲酸。
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