基于平衡UDP-糖供给合成乳酰-N-新四糖的重组枯草芽孢杆菌及其构建方法与应用与流程

本发明属于遗传工程的技术领域，具体涉及一种基于平衡udp-糖供给合成乳酰-n-新四糖的重组枯草芽孢杆菌及其构建方法与应用。

背景技术：

乳酰-n-新四糖是母乳中是重要的寡糖成分之一，由于具有促进细胞分化、提高肠道上皮细胞免疫调节功能、增加肠道益生菌的相对丰度等重要生理功能，被欧洲食品安全局(efsa)和美国的fda批准为食品天添加剂。到目前为止，仅在人乳中发现乳酰-n-新四糖，而从人乳中提取并不适合大规模生产。由于化学合成步骤繁琐、转化率低，酶法合成存在底物价格昂贵的缺点，所以采用绿色环境友好型的生物合成有望实现乳酰-n-新四糖的工业化生产。

枯草芽孢杆菌(bacillussubtilis)作为革兰氏阳性菌中的一种代表模式菌株，并且不产生内毒素，被美国食品药品监督管理局(fda)认证为是(generallyrecognizedassafe，gras)。因此，运用代谢工程手段构建枯草芽孢杆菌工程菌是生产乳酰-n-新四糖的有效策略。其自身途径中的尿苷二磷酸半乳糖，乙酰氨基葡萄糖尿苷二磷酸是合成的关键前体，但是由于这两个udp-糖的合成途径中的关键酶催化活力不足，以及前体之间供给的不平衡限制了产物的高效合成。因此，如何对前体udp-糖的供给进行平衡是亟需解决的问题。

技术实现要素：

为了解决上述问题，本发明提供了一种基于平衡udp-糖供给合成乳酰-n-新四糖的重组枯草芽孢杆菌及其构建方法与应用。

具体的，一方面，本发明提供了一种基于平衡udp-糖供给合成乳酰-n-新四糖的重组枯草芽孢杆菌，所述重组枯草芽孢杆菌是通过在宿主菌bacillussubtilis168△amye:p43-lacy，p43-lgtb，pxyla-comk的基因组上整合两个β-1,4-半乳糖转移酶，并整合葡萄糖-6-磷酸异构酶基因、谷氨酰胺-果糖-6-磷酸氨基转移酶基因、utp-葡萄糖-1-磷酸尿苷酰转移酶基因、udp-葡萄糖-差向异构酶基因，且敲除udp-葡萄糖6-脱氢酶基因，并外源表达β-1,3-n-葡糖氨基转移酶基因得到的。

本发明通过强化表达4个关键酶基因葡萄糖-6-磷酸异构酶基因、谷氨酰胺-果糖-6-磷酸氨基转移酶基因、utp-葡萄糖-1-磷酸尿苷酰转移酶基因、udp-葡萄糖-差向异构酶基因，以及敲除1个分支途径关键酶udp-葡萄糖6-脱氢酶基因，即通过优化udp-糖前体供给的合成途径中关键酶基因的表达量以及敲除分支途径中关键酶基因，明显增加乳酰-n-新四糖的产量。

需要进一步说明的，所述宿主菌是以枯草芽孢杆菌168为基础，通过以启动子pxyla调控基因comk的表达，乳糖透性酶编码基因整合到基因组amye位点，β-1,4-半乳糖转移酶编码基因整合到基因组ydes位点和ydzo位点中间得到的。

第二方面，本发明还提供了所述的基于平衡udp-糖供给合成乳酰-n-新四糖的构建方法，该方法包括以下步骤：

(1)通过融合pcr构建包含ydah，ydza基因同源臂、β-1,4-半乳糖转移酶基因、p43启动子、博来霉素抗性基因序列的重组片段1；包含ysza，ysxe基因同源臂、β-1,4-半乳糖转移酶基因、p43启动子、氯霉素抗性基因序列的重组片段2；

(2)通过融合pcr构建p43控制表达的葡萄糖-6-磷酸异构酶基因的重组片段3，p43控制表达的谷氨酰胺-果糖-6-磷酸氨基转移酶基因的重组片段4；

(3)通过融合pcr构建p43控制表达的utp-葡萄糖-1-磷酸尿苷酰转移酶基因的重组片段5，p43控制表达的udp-葡萄糖-差向异构酶基因的重组片段6，包含tuad基因左同源臂片段，p7z6片段，tuad基因右同源臂片段的重组片段7；

(4)将步骤(1)构建的重组片段1转化宿主菌，通过验证获得重组枯草芽孢杆菌by04；再将重组片段转化重组枯草芽孢杆菌by04，得到重组枯草芽孢杆菌by05；

(5)将步骤(2)构建的重组片段3转化重组枯草芽孢杆菌by05，获得重组枯草芽孢杆菌mi1，再将重组片段4转化mi1，得到重组枯草芽孢杆菌mi2；将mi2菌进行抗性基因的消除，得到mi2.2菌；

(6)将步骤(3)构建的重组片段5转化mi2.2菌，得到重组枯草芽孢杆菌mi3，再将重组片段6转化步骤mi3菌，得到重组枯草芽孢杆菌mi4，再将重组片段7转化mi4菌，得到重组枯草芽孢杆菌mi5.0；

(7)将pp43nmk-lgta转化重组枯草芽孢杆菌mi5.0，得到合成乳酰-n-新四糖的重组枯草芽孢杆菌mi5。

进一步的，步骤(1)中，重组片段1的序列如seqidno.1所示，重组片段2的序列如seqidno.2所示。

进一步的，步骤(2)中，重组片段3的序列如seqidno.3所示，重组片段4的序列如seqidno.4所示。

进一步的，步骤(3)中，重组片段5的序列如seqidno.5所示，重组片段6的序列如seqidno.6所示，重组片段7的序列如seqidno.7所示。

进一步的，步骤(5)中，通过同源重组，葡萄糖-6-磷酸异构酶基因插入在rsbrc位点和yojg位点之间，谷氨酰胺-果糖-6-磷酸氨基转移酶基因插入在comer位点和yqem位点之间。

进一步的，步骤(6)中，通过同源重组，utp-葡萄糖-1-磷酸尿苷酰转移酶基因插入在yyca位点和yybt位点之间，udp-葡萄糖-差向异构酶基因插入在ybge位点和ybgf位点之间。

第三方面，本发明还提供了所述重组枯草芽孢杆菌在发酵生产乳酰-n-新四糖中的应用。

本发明的有益效果为：

本发明通过优化udp-糖前体供给的合成途径中关键酶基因的表达量以及敲除分支途径中关键酶基因，明显增加乳酰-n-新四糖的产量。本发明构建的重组枯草芽孢杆菌合成乳酰-n-新四糖，在摇瓶上产量可达到1800mg/l，实现了乳酰-n-新四糖的高效合成，为进一步代谢工程改造枯草芽孢杆菌生产乳酰-n-新四糖及其工业化奠定了基础。

附图说明

图1为本发明重组mi1菌株的pcr琼脂糖凝胶电泳图；

图2为本发明重组mi2菌株的pcr琼脂糖凝胶电泳图；

图3为本发明重组mi3菌株的pcr琼脂糖凝胶电泳图；

图4为本发明重组mi4菌株的pcr琼脂糖凝胶电泳图；

图5为本发明重组mi5.0菌株的pcr琼脂糖凝胶电泳图；

图6为平衡udp-糖供给后对乳酰-n-新四糖产量的影响；

图7为不同udp-gal强化程度对乳酰-n-新四糖产量的影响。

具体实施方式

下面结合实施例和附图对本发明进行详细的解释说明。

实施例1重组pzl(p43-lacy)片段的构建

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，根据ncbi上公布的amye基因(geneid：938356)，设计两侧同源臂引物，序列分别为seqidno:1和seqidno:2的左侧同源臂引物：

amye-1f:5’-tattccgtatgtcaagtggctgcggtttat-3’(seqidno:1)，

amye-1r:5’-aattgttatccgctctcttgacactccttatttgattttttgaagacttacttcgg-3’(seqidno:2)，

序列分别为seqidno:3和seqidno:4的右侧同源臂引物：

amye-2f:5’-cttaagggcaaggctagacgggactta-3’(seqidno:3)，

amye-2r:5’-ggcacaccgatgtacacgtcatc-3’(seqidno:4)，

使用上述引物从枯草芽孢杆菌基因组中扩增amye两侧同源臂基因序列；以质粒pp43nmk为模板，分别设计序列为seqidno:5和seqidno:6的引物：

p43-f:5’-cattatacgaacggtaaatctgataggtggtatgttttcgcttgaacttttaaatacag-3’(seqidno:5)，

p43-r:5’-agtttgtgttttttaaatagtacatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:6)，

使用上述引物，获得p43的扩增片段。以p7z6质粒为模板，设计序列分别为seqidno:7和seqidno:8的博来霉素抗性基因扩增引物：

zeo-f:5’-tcaaataaggagtgtcaagagagcggataacaatttcacacaggaaacag-3’(seqidno:7)，

zeo-r:5’-aacataccacctatcagatttaccgttcgtataatgtatgctatacgaagttattcagt-3’(seqidno:8)，

使用上述引物，获得博来霉素抗性基因的扩增片段。将p43的扩增片段和博来霉素抗性基因扩增片段跑胶验证正确后进行柱回收。将p43的扩增片段和博来霉素抗性基因扩增片段通过融合pcr技术进行融合。第一轮pcr条件为：等摩尔量柱回收的p43和博来霉素抗性dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，zeo-f，p43-r分别为上下游引物，按照常规pcr设置条件，获得zp43片段。

以大肠杆菌(escherichiacolik-12)基因组为模板，根据ncbi上公布的乳糖透性酶基因lacy(geneid：949083)，设计序列分别为seqidno:9和seqidno:10的引物：

lacy-f:5’-gtaagagaggaatgtacacatgtactatttaaaaaacacaaacttttggatgttcggtt-3’(seqidno:9)，

lacy-r:5’-taagtcccgtctagccttgcccttaagcgacttcattcacctgacgacg-3’(seqidno:10)，

使用上述引物扩增乳糖透性酶基因片段。将amye基因左侧同源臂、amye基因右侧同源臂、zp43片段、乳糖透性酶基因片段跑胶验证正确后进行柱回收。通过融合pcr技术将amye基因左侧同源臂、amye基因右侧同源臂、zp43片段、乳糖透性酶基因片段进行融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:11和seqidno:12所示的序列:

zong-f:5’-catgtaagccataagccattcgtaa-3’(seqidno:11)，

zong-r:5’-agcaagactcatcgcaaccc-3’(seqidno:12)分别为上下游引物，按照常规pcr设置条件，获得pzl片段。重组片段pzl的序列如seqidno:13所示。

实施例2重组pxyla-comk片段的构建

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:14和seqidno:15的左侧同源臂引物：

yhzc-f:5’-catacataggaagcaggcattgttcataac-3’(seqidno:14)，

yhzc-r:5’-atacgggatcaaatccgatgaaagagaaaaaatcgtacactgagctc-3’(seqidno:15)，

序列分别为seqidno:16和seqidno:17的右侧同源臂引物：

comk-f:5’-aagggggaaatgggatccatgagtcagaaaacagacgcacct-3’(seqidno:16)，

comk-r:5’-actacctcagttgaaggctataatccaag-3’(seqidno:17)，

使用上述引物从枯草芽孢杆菌基因组中两侧同源臂基因序列；以质粒plcx-dcas9为模板，分别设计序列为seqidno:18和seqidno:19的引物：

pxyla-f:5’-tttctctttcatcggatttgatcccgtataccgttcgtatagcatacattat-3’(seqidno:18)，

pxyla-r:5’-gactcatggatcccatttccccctttgatttttagatatcac-3’(seqidno:19)，

使用上述引物，获得带有氯霉素抗性的pxyla启动子的扩增片段。通过融合pcr，将重组同源臂以及带抗性基因的启动子融合，第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:20和seqidno:21所示的序列:

zong-f:5’-gacccgcgtccggtttttgataagccgaagaaag-3’(seqidno:20)，

zong-r:5’-agtaaattcagttgcgcccggaccaatttggacac-3’(seqidno:21)，

分别为上下游引物，按照常规pcr设置条件，获得pxyla-comk片段，片段pxyla-comk的序列如seqidno:22所示。

实施例3p7s6p43-lgtb片段的构建

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:23和seqidno:24的左侧同源臂引物：

ydes-f:5’-gggacaaggaatagtaagccggcaa-3’(seqidno:23)，

ydes-r:5’-tcctgtgtgaaattgttatccgctcctacatactctctgtagcagaggtagcttga’(seqidno:24)，

序列分别为seqidno:25和seqidno:26的右侧同源臂引物：

ydzo-f:5’-tgaagcccgcctaatgagcgggcttttttctgataagaactgcaaaagctgcggattat-3’(seqidno:25)，

ydzo-r:5’-ccaccctatagataaatttttcggctgccatat-3’(seqidno:26)，

使用上述引物从枯草芽孢杆菌基因组中扩增两侧同源臂基因序列。

以质粒p7s6p43为模板，分别设计序列为seqidno:27和seqidno:28的引物：

p7s6p43-f:5’-agagtatgtaggagcggataacaatttcacacagga-3’(seqidno:27)，

p7s6p43-r:5’-aaagaaatgacatgattttgcatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:28)，

以重组质粒pp43nmk-lgta-lgtb为模板(在专利cn108410787a中已经有公开)，分别设计序列为seqidno:29和seqidno:30的引物：

lgtb-f:5’-aggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagcag-3’(seqidno:29)，

lgtb-r:5’-cccgctcattaggcgggcttcatcattactgaaacggaacgataaactgttcgcg-3’(seqidno:30)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将重组同源臂，β-1,4-半乳糖转移酶编码基因以及带抗性基因的启动子融合，第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:31和seqidno:32所示的序列:

zong-f:5’-cgtatccgatttgggtgagtgtga-3’(seqidno:31)，

zong-r:5’-tgatgtcaatctaatgcctccttactgg-3’(seqidno:32)分别为上下游引物，按照常规pcr设置条件，获得p7s6p43-lgtb片段。p7s6p43-lgtb片段的序列如seqidno:33所示。

实施例4重组质粒pp43nmk-lgta的构建

以重组质粒pp43nmk-lgta-lgtb为模版，设计反向重组引物，序列分别为seqidno:34和seqidno:35：

fz-lgta-f:5’-tgaagcccgcctaatgagcgggcttttttaagcttggcgtaatcatggtcatagctgtt-3’(seqidno:34)，

fz-lgta-r:5’-ctgtgtgaaattgttatccgctcccagcctttcttattaaaaccactttgtcagcc-3’(seqidno:35)，

使用上述引物将获得的扩增片段跑胶验证正确后，取10μl的pcr反应体系，转化大肠杆菌jm109感受态，通过测序，验证重组质粒pp43nmk-lgta构建成功，重组质粒pp43nmk-lgta的基因序列如seqidno:36所示。

实施例5重组p7z6p43-lgtb片段的构建

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:37和seqidno:38的左侧同源臂引物：

ydah-f:5’-ggctttcgttgtcttgtgttcaagaaatttcca-3’(seqidno:37)，

ydah-r:5’-ctgtgtgaaattgttatccgctcccagcctttcttattaaaaccactttgtcagcc-3’(seqidno:38)，

序列分别为seqidno:39和seqidno:40的右侧同源臂引物：

ydza-f:5’-aagcccgcctaatgagcgggcttttttatcataaagatccagcctttttgcgct-3’(seqidno:39)，

ydza-r:5’-ctaaagcccaagtcacaatatattgatcgcct-3’(seqidno:40)，

以质粒p7z6p43为模板，分别设计序列为seqidno:41和seqidno:42的引物：

p7z6p43-f:5’-gttttaataagaaaggctgggagcggataacaatttcacacaggaaacagc-3’(seqidno:41)，

p7z6p43-r:5’-aaagaaatgacatgattttgcatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:42)，

以出发菌株bacillussubtilis168△amye:p43-lacy，p43-lgtb，pxyla-comk的基因组为模板，分别设计序列为seqidno:43和seqidno:44的引物：

lgtb-1f:5’-aggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagcag-3’(seqidno:43)，

lgtb-1r:5’-cccgctcattaggcgggcttcatcattactgaaacggaacgataaactgttcgcg-3’(seqidno:44)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将重组同源臂，β-1,4-半乳糖转移酶编码基因以及带抗性基因的启动子融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，融合pcr的引物分别为seqidno:45和seqidno:46：

zong-1f:5’-aacctccttgtcttctttccagtcttatctc-3’(seqidno:45)，

zong-1r:5’-atcacttctctttctttcacgctcatcct-3’(seqidno:46)，按照常规pcr设置条件，获得p7z6p43-lgtb片段，p7z6p43-lgtb片段的序列如seqidno:47所示。

实施例6重组p7c6p43-lgtb片段的构建

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:48和seqidno:49的左侧同源臂引物：

ysza-f:5’-catccagcgtaaaacgttcacgggaataatctagg-3’(seqidno:48)，

ysza-r:5’-ttatccgctctggcgcggacttgtttgtttatatccattctaaatgaagg-3’(seqidno:49)，

序列分别为seqidno:50和seqidno:51的右侧同源臂引物：

ysxe-f:5’-gcccgcctaatgagcgggcttttttttaagacgtggactcgttttcagcctgaaatttt-3’(seqidno:50)，

ysxe-r:5’-catcccagcagctgatcaggatgaattct-3’(seqidno:51)，

以质粒p7c6p43为模板，分别设计序列为seqidno:52和seqidno:53的引物：

p7c6p43-f:5’-acaagtccgcgccagagcggataacaatttcacacaggaaacagctatg-3’(seqidno:52)，

p7c6p43-r:5’-agaaatgacatgattttgcatgtgtacattcctctcttacctataatggtaccgct-3’(seqidno:53)，

以出发菌株基因组为模板，分别设计序列为seqidno:54和seqidno:55的引物：

lgtb-2f:5’-ataggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagcagcg-3’(seqidno:54)，

lgtb-2r:5’-gcccgctcattaggcgggcttcatcattactgaaacggaacgataaactgttcgcgt-3’(seqidno:55)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将重组同源臂，β-1,4-半乳糖转移酶编码基因以及带抗性基因的启动子融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，融合pcr的引物分别为seqidno:56和seqidno:57：

zong-2f:5’-gccaggaagccacagcacatcatagc-3’(seqidno:56)，

zong-2r:5’-ttcgttcattcgttcccgtttacagaacaaatagc-3’(seqidno:57)，按照常规pcr设置条件，获得p7c6p43-lgtb片段，p7c6p43-lgtb片段的序列如seqidno:58所示。

实施例7出发菌株枯草芽孢杆菌的构建

将构建好的重组片段pxyla-comk转化枯草芽孢杆菌感受态细胞(bacillussubtilis168)，重组片段的添加量为100-300ng，电转条件：电压2.5kv，电击试剂5ms，37℃复苏5h后涂布氯霉素抗性平板，37℃培养24h，获得重组枯草芽孢杆菌bs168comk。

bs168comk重组枯草芽孢杆菌通过木糖诱导制作感受态。在37℃，220rpm条件下，lb液体培养12h后，用lb液体培养基将od600稀释至1，并添加终浓度为2％的木糖，继续培养2h，获得bs168comk感受态，重组片段pzl的添加量为500ng，37℃培养1h后涂布博来霉素抗性平板，37℃培养过夜，获得重组枯草芽孢杆菌by00。

by00通过木糖诱导制作成感受态细胞，重组片段p7s6p43-lgtb的添加量为500ng，37℃培养1h后涂布壮观霉素抗性平板，37℃培养过夜，获得重组枯草芽孢杆菌by01。通过cre/lox重组系统，敲除抗性基因，获得by01.2菌。

by01.2通过木糖诱导制作成感受态细胞，重组质粒pp43nmk-lgta的添加量为500ng，37℃培养1h后涂布卡那霉素抗性平板，37℃培养过夜，获得菌株by03。

实施例8

将构建好的重组p7z6p43-lgtb片段转化感受态细胞by01.2，挑选转化子进行菌落pcr验证，分别设计验证引物序列seqidno:59和seqidno:60的引物：

yz-1f:5’-cgaaggaatgccggtgtgccaactgaatc-3’(seqidno:59)，

yz-1r:5’-cgacctccagcgtgtattgcttgatgatgc-3’(seqidno:60)，验证正确后获得重组枯草芽孢杆菌by03a，菌落pcr琼脂糖凝胶电泳图如图1所示。

将重组质粒pp43nmk-lgta转化上述重组枯草芽孢杆菌by03a，37℃培养1h后涂布卡那霉素抗性平板，37℃培养过夜，获得重组枯草芽孢杆菌by04。

将构建好的重组p7c6p43-lgtb片段转化重组枯草芽孢杆菌by03a感受态细胞，挑选转化子进行菌落pcr验证，分别设计验证引物序列seqidno:61和seqidno:62的引物：

yz-2f:5’-gggcacagcgggatacatcatctcaagaaa-3’(seqidno:61)，

yz-2r:5’-tgaaccacggcagcctgtctatacaccat-3’(seqidno:62)。

验证正确后获得重组枯草芽孢杆菌by03b，得到的菌落pcr琼脂糖凝胶电泳图如图2所示。

将重组质粒pp43nmk-lgta转化重组枯草芽孢杆菌by03b，37℃培养1h后涂布卡那霉素抗性平板，37℃培养过夜，获得一种提高乳酰-n-新四糖的重组枯草芽孢杆菌by05。

所用plcx-dcas9，p7s6p43，p7z6p43，p7c6p43质粒来自江苏省无锡市滨湖区蠡湖大道1800号，生物工程学院，生物系统与生物加工工程研究室，上述p7s6p43，p7z6p43，p7c6p43质粒在modularpathwayengineeringofkeycarbon-precursorsupply-pathwaysforimprovedn-acetylnuraminicacidproductioninbacillussubtilis(zhangxl，liuyf，liul，wangm，lijh，dugc，chenj.2018)中已经公开。上述plcx-dcas9质粒在crispriallowsoptimaltemporalcontrolofnacetylglucosaminebioproductionbyadynamiccoordinationofglucoseandxylosemetabolisminbacillussubtilis(wuyk,chentc,liuyf,lvxq,lijh，dugc，amarorl,liul.2018)中已经公开。

实施例9udp-glcnac前体供给的强化

本发明中的udp-glcnac的前体供给强化包括强化途径中葡萄糖-6-磷酸异构酶基因pgi，谷氨酰胺-果糖-6-磷酸氨基转移酶基因glms。

葡萄糖-6-磷酸异构酶基因pgi的强化，其构建过程如下：

设计将p43-pgi基因插入在rsbrc位点和yoji位点之间

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:63和seqidno:64的左侧同源臂引物：

rsbrc-f:5’-gagggtacaaagatgtgaactatacgcttgc_-3’(seqidno:63)，

rsbrc-r:5’-gaaattgttatccgctcaaggccttaagccttataccttttttaattttaagtccga-3’(seqidno:64)，

序列分别为seqidno:65和seqidno:66的右侧同源臂引物：

yogi-f:5’-gcccgcctaatgagcgggcttttttatacctttttgattaatcattccattgatacgtcca-3’(seqidno:65)，

yogi-r:5’-catgaaagcgatcataaaagaagatgtgcaag-3’(seqidno:66)，

以质粒p7z6p43为模板，分别设计序列为seqidno:67和seqidno:68的引物：

p7z6p43-f:5’-ggtataaggcttaaggccttgagcggataacaatttcacacaggaaacag-3’(seqidno:67)，

p7z6p43-r:5’-caaagcgtacatgcgtcatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:68)，

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，分别设计序列为seqidno:69和seqidno:70的引物：

pgi-f:5’-agaggaatgtacacatgacgcatgtacgctttgactactcaaaa-3’(seqidno:69)，

pgi-r:5’-gcccgctcattaggcgggcttcatcattaatcttccagacgtttttcaagctctgctt-3’(seqidno:70)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将左同源臂片段，p7zp43片段，pgi片段，右同源臂片段融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:71和seqidno:72所示的序列:

zong-f:5’-gctgtttgcgttcttgccgtgatttcgtgta-3’(seqidno:71)，

zong-r:5’-gataaatgcccttctcatctttcccttcgctttctcc-3’(seqidno:72)分别为上下游引物，按照常规pcr设置条件，获得3900bp的扩增片段，如seqidno.73所示。

谷氨酰胺-果糖-6-磷酸氨基转移酶基因glms的强化，设计如下：

设计将p43-glms基因插入在comer位点和yqem位点之间。

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:74和seqidno:75的左侧同源臂引物：

comer-f:5’-gagggtacaaagatgtgaactatacgcttgc_-3’(seqidno:74)，

comer-r:5’-gaaattgttatccgctcaaggccttaagccttataccttttttaattttaagtccga-3’(seqidno:75)，

序列分别为seqidno:76和seqidno:77的右侧同源臂引物：yqem-f:5’-taaagcccgcctaatgagcgggcttttttacgagtatgtcatacgttcgtctgtactgcaaaa-3’(seqidno:76)，

yqem-r:5’-ccattttaaagatcgcagctgcggc-3’(seqidno:77)，

以质粒p7s6p43为模板，分别设计序列为seqidno:78和seqidno:79的引物：

p7s6p43-f:5’-agcagtttcatgtgtaggagcggataacaatttcacacaggaaacag-3’(seqidno:78)，

p7s6p43-r:5’-acctacgattccacacatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:79)，

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，分别设计序列为seqidno:80和seqidno:81的引物：

glms-f:5’-agaggaatgtacacatgtgtggaatcgtaggttatatcggtcagct-3’(seqidno:80)，no:80)，

glms-r:5’-cccgctcattaggcgggctttactccacagtaacactcttcgcaaggttacga-3’(seqidno:81)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将左同源臂片段，p7sp43片段，glms片段，右同源臂片段融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:82和seqidno:83所示的序列:

zong-f:5’-tgtttcgtttgactcgtcgttgtttgcttcct-3’(seqidno:82)，

zong-r:5’-ctaggcgatgtagttgtccatgtcttccacaag-3’(seqidno:83)分别为上下游引物，按照常规pcr设置条件，获得5335bp的扩增片段，如seqidno.84所示。

重组枯草芽孢杆菌(bacillussubtilis168△amye::p43-lacy,3copiesp43-lgtb,pxyla-comk)by03b为宿主菌，通过cre/lox重组系统，敲除抗性基因，获得by03b.2。消除抗性基因后，葡萄糖-6-磷酸异构酶基因pgi的扩增片段的添加量为500ng，37℃培养1h后涂布博来霉素抗性平板，37℃培养过夜，获得mi1菌株，菌落pcr琼脂凝胶电泳图如图1所示。

谷氨酰胺-果糖-6-磷酸氨基转移酶基因glms的扩增片段的添加量为500ng，转化到重组枯草芽孢杆菌mi1感受态细胞，37℃培养1h后涂布壮观霉素抗性平板，37℃培养过夜，获得mi2菌株。通过cre/lox重组系统，敲除抗性基因，获得mi2.2，菌落pcr琼脂凝胶电泳图如图2所示。

实施例10udp-gal前体供给的强化

本发明中的udp-gal的前体供给强化包括强化途径中utp-葡萄糖-1-磷酸尿苷酰转移酶基因gtab，udp-葡萄糖-差向异构酶基因gale以及敲除udp-葡萄糖-脱氢酶基因tuad。

utp-葡萄糖-1-磷酸尿苷酰转移酶基因gtab的强化，其构建过程如下：

设计将p43-gtab基因插入在yyca位点和yybt位点之间。

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:85和seqidno:86的左侧同源臂引物：

yyca-f:5’-ggcaacggttcatctttttctcagaacg_-3’(seqidno:85)，

yyca-r:5’-cctgtgtgaaattgttatccgctcttattccacgagataaagagtttgctggct-3’(seqidno:86)，

序列分别为seqidno:87和seqidno:88的右侧同源臂引物：yybt-f:5’-gcccgcctaatgagcgggcttttttatacctttttgattaatcattccattgatacgtcca-3’(seqidno:87)，

yybt-r:5’-catgaaagcgatcataaaagaagatgtgcaag-3’(seqidno:88)，

以质粒p7s6p43为模板，分别设计序列为seqidno:89和seqidno:90的引物：

p7s6p43-f:5’-ctttatctcgtggaataagagcggataacaatttcacacaggaaacag-3’(seqidno:89)，

p7s6p43-r:5’-caaagcgtacatgcgtcatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:90)，

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，分别设计序列为seqidno:91和seqidno:92的引物：

gtab-f:5’-aggtaagagaggaatgtacacatgaaaaaagtacgtaaagccataattccagca-3’(seqidno:91)，

gtab--r:5’-gcccgctcattaggcgggctttagatttcttctttgtttagtaaaccttccataaatggaacg-3’(seqidno:92)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将左同源臂片段，p7sp43片段，gtab片段，右同源臂片段融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:93和seqidno:94所示的序列:

zong-f:5’-ttcttctattatcggtgttcagcaggt-3’(seqidno:93)，

zong-r:5’-ggaggcgtacagagatgaaggttat-3’(seqidno:94)分别为上下游引物，按照常规pcr设置条件，获得4509bp的扩增片段，如seqidno.95所示。

udp-葡萄糖-差向异构酶基因gale的强化，其构建过程如下：设计将p43-gale基因插入在ybge位点和ybgf位点之间。

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:96和seqidno:97的左侧同源臂引物：

ybge-f:5’-gtgaagcggcgtattattaaaggaaaggctgta_-3’(seqidno:96)，

ybge-r:5’-tgtgtgaaattgttatccgctctcacacttccactgtccagttaaacgggc-3’(seqidno:97)，

序列分别为seqidno:98和seqidno:99的右侧同源臂引物：

ybgf-f:5’-agcccgcctaatgagcgggcttttttaaatcgaaaaagaacctgcccggaggca-3’(seqidno:98)，

ybgf-r:5’-ccggatttacggtcgcttggctgtatt-3’(seqidno:99)，

以质粒p7s6p43为模板，分别设计序列为seqidno:100和seqidno:101的引物：

p7c6p43-f:5’-cagtggaagtgtgagagcggataacaatttcacacaggaaacag-3’(seqidno:100)，

p7c6p43-r:5’-gccagtaacaagtattgccatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:101)，

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，分别设计序列为seqidno:102和seqidno:103的引物：

gale-f:5’-aagagaggaatgtacacatggcaatacttgttactggcggt-3’(seqidno:102)，

gale-r:5’-cccgctcattaggcgggctttattccgcactcttatacccattcacattagaaga-3’(seqidno:103)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将左同源臂片段，p7cp43片段，gale片段，右同源臂片段融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:104和seqidno:105所示的序列:

zong-f:5’-aaagcccgacccgtcttcct-3’(seqidno:104)，

zong-r:5’-ttggatgtggagtgccgtctt-3’(seqidno:105)分别为上下游引物，按照常规pcr设置条件，获得4666bp的扩增片段，如seqidno.106所示。

udp-葡萄糖-脱氢酶基因tuad的敲除，其构建过程如下：

以枯草芽孢杆菌(bacillussubtilis168)基因组为模板，设计两侧同源臂引物，序列分别为seqidno:107和seqidno:108的左侧同源臂引物：

tuac-f:5’-caaattccttatgtcctgaccttgcatggc_-3’(seqidno:107)，

tuac-r:5’-tgtgtgaaattgttatccgctcagcgctcaaccctctcctgtaaaat_-3’(seqidno:108)，

序列分别为seqidno:109和seqidno:110的右侧同源臂引物：

tuae-f:5’-accctggcgttacggaaccctctgacaagtattttccgg-3’(seqidno:109)，

tuae-r:5’-gccattctctttaaaacaggcggcaatacaa-3’(seqidno:110)，

以质粒p7z6为模板，分别设计序列为seqidno:111和seqidno:112的引物：

p7z6-f:5’-cagtggaagtgtgagagcggataacaatttcacacaggaaacag-3’(seqidno:111)，

p7z6-r:5’-gccagtaacaagtattgccatgtgtacattcctctcttacctataatggtaccgc-3’(seqidno:112)，

使用上述引物，将获得的扩增片段跑胶验证正确后进行柱回收。通过融合pcr，将左同源臂片段，p7z6片段，右同源臂片段融合。第一轮pcr条件为：等摩尔量柱回收的dna，总量大于1000ng，primerstar酶量25μl，加ddh2o定容50μl，pcr条件55℃，11个循环。第二轮pcr条件为：上述pcr产物为模板，以seqidno:113和seqidno:114所示的序列：

zong-f:5’-ccgtaggttctgcgtcagttgtt-3’(seqidno:113)，

zong-r:5’-gatgtcggataatgctgcttgtactc-3’(seqidno:114)分别为上下游引物，按照常规pcr设置条件，获得2238bp的扩增片段，如seqidno.115)。

utp-葡萄糖-1-磷酸尿苷酰转移酶基因gtab的扩增片段的添加量为500ng，转化到重组枯草芽孢杆菌mi2.2感受态细胞，37℃培养1h后涂布壮观霉素抗性平板，37℃培养过夜，获得mi3菌株，菌落pcr琼脂凝胶电泳图如图3所示。

udp-葡萄糖-差向异构酶基因gale的扩增片段添加量为500ng，转化到重组枯草芽孢杆菌mi3感受态细胞，37℃培养1h后涂布氯霉素抗性平板，37℃培养过夜，获得mi4菌株，菌落pcr琼脂凝胶电泳图如图4所示。

敲除udp-葡萄糖-脱氢酶基因tuad的扩增片段添加量为500ng，转化到重组枯草芽孢杆菌mi4感受态细胞，37℃培养1h后涂布博来霉素抗性平板，37℃培养过夜，获得mi5.0菌株，菌落pcr琼脂凝胶电泳图如图5所示。

将重组质粒pp43nmk-lgta转化重组枯草芽孢杆菌mi5.0，37℃培养1h后涂布卡那霉素抗性平板，37℃培养过夜，获得一种提高乳酰-n-新四糖的重组枯草芽孢杆菌mi5。

实施例11重组枯草芽孢杆菌mi5发酵法生产乳酰-n-新四糖

采用实施例10构建得到的重组枯草芽孢杆菌mi5菌与宿主菌进行摇瓶发酵，制成种子液。种子液的制作方法为：挑取平板上的单菌落在lb液体培养基中，同时添加终浓度为30μg/ml的卡那霉素，在37℃、220rpm条件下，培养10-12h。

将种子液按10％的接种量转入发酵培养基中(发酵培养基：胰蛋白胨6g/l，酵母粉12g/l，(nh4)so46g/l，k2hpo4·3h2o12.5g/l，kh2po42.5g/l，mgso4·7h2o1.2g/l，葡萄糖60g/l，乳糖5g/l)，于37℃、220rpm条件下培养48h。如图6在发酵培养液的上清测得乳酰-n-新四糖，含量为1800mg/l。较重组枯草芽孢杆菌by05提高38.5％；同时本发明提供的重组枯草芽孢杆菌发酵产乳酰-n-新四糖的葡萄糖得率为33.7mg/g，较重组枯草芽孢杆菌by05提高了25％，实现了乳酰-n-新四糖在重组枯草芽孢杆菌的生产强度和产量的提高，为进一步代谢工程改造枯草芽孢杆菌生产乳酰-n-新四糖及其工业化奠定了基础。

对照实施例1

pdg-cre质粒用于敲除mi1菌株的抗性基因，获得无抗性的mi1.2菌株。重组枯草芽孢杆菌mi1.2为宿主菌，将utp-葡萄糖-1-磷酸尿苷酰转移酶基因gtab的扩增片段转化到宿主菌，获得mi6.0菌株。

将udp-葡萄糖-差向异构酶基因gale的扩增片段转化到重组枯草芽孢杆菌mi6.0菌株，获得mi7.0菌株。

将敲除udp-葡萄糖-脱氢酶基因tuad的扩增片段转化到重组枯草芽孢杆菌mi7.0菌株，获得mi8.0菌株。

将重组质粒pp43nmk-lgta转化重组枯草芽孢杆菌mi6.0、mi7.0、mi8.0，37℃培养1h后涂布卡那霉素抗性平板，37℃培养过夜，分别获得重组枯草芽孢杆菌mi6、mi7、mi8。

将mi6、mi7、mi8进行摇瓶发酵。将三株菌制成种子液，种子液的制作方法为：挑取平板上的单菌落在lb液体培养基中，同时添加终浓度为30μg/ml的卡那霉素，在37℃、220rpm条件下，培养10-12h。将种子液按10％的接种量转入发酵培养基中。于37℃、220rpm条件下培养48h。在发酵培养液的上清测得乳酰-n-新四糖。已知mi6、mi7、mi8与mi3、mi4、mi5相对应的都没有强化谷氨酰胺-果糖-6-磷酸氨基转移酶基因glms。如图7可知，mi6、mi7、mi8菌株均没有mi5菌产乳酰-n-新四糖的含量高。

以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明实质内容上所作的任何修改、等同替换和简单改进等，均应包含在本发明的保护范围之内。

序列表

<110>光明乳业股份有限公司

<120>基于平衡udp-糖供给合成乳酰-n-新四糖的重组枯草芽孢杆菌及其构建方法与应用

<160>115

<170>siposequencelisting1.0

<210>1

<211>30

<212>dna

<213>人工序列(未知)

<400>1

tattccgtatgtcaagtggctgcggtttat30

<210>2

<211>56

<212>dna

<213>人工序列(未知)

<400>2

aattgttatccgctctcttgacactccttatttgattttttgaagacttacttcgg56

<210>3

<211>27

<212>dna

<213>人工序列(未知)

<400>3

cttaagggcaaggctagacgggactta27

<210>4

<211>23

<212>dna

<213>人工序列(未知)

<400>4

ggcacaccgatgtacacgtcatc23

<210>5

<211>59

<212>dna

<213>人工序列(未知)

<400>5

cattatacgaacggtaaatctgataggtggtatgttttcgcttgaacttttaaatacag59

<210>6

<211>59

<212>dna

<213>人工序列(未知)

<400>6

agtttgtgttttttaaatagtacatgtgtacattcctctcttacctataatggtaccgc59

<210>7

<211>50

<212>dna

<213>人工序列(未知)

<400>7

tcaaataaggagtgtcaagagagcggataacaatttcacacaggaaacag50

<210>8

<211>59

<212>dna

<213>人工序列(未知)

<400>8

aacataccacctatcagatttaccgttcgtataatgtatgctatacgaagttattcagt59

<210>9

<211>59

<212>dna

<213>人工序列(未知)

<400>9

gtaagagaggaatgtacacatgtactatttaaaaaacacaaacttttggatgttcggtt59

<210>10

<211>49

<212>dna

<213>人工序列(未知)

<400>10

taagtcccgtctagccttgcccttaagcgacttcattcacctgacgacg49

<210>11

<211>25

<212>dna

<213>人工序列(未知)

<400>11

catgtaagccataagccattcgtaa25

<210>12

<211>20

<212>dna

<213>人工序列(未知)

<400>12

agcaagactcatcgcaaccc20

<210>13

<211>3811

<212>dna

<213>人工序列(未知)

<400>13

catgtaagccataagccattcgtaaaagtgcgggaggaaggtcatgaataatctgcgtaa60

tagactttcaggcgtgaatgggaaaaataagagagtaaaagaaaaagaacaaaaaatctg120

gtcggagattgggatgatagcgggagcatttgcgctgcttgatgtgatcatccgcggcat180

tatgtttgaatttccgtttaaagaatgggctgcaagccttgtgtttttgttcatcattat240

cttatattactgcatcagggctgcggcatccggaatgctcatgccgagaatagacaccaa300

agaagaactgcaaaaacgggtgaagcagcagcgaatagaatcaattgcggtcgcctttgc360

ggtagtggtgcttacgatgtacgacagggggattccccatacattcttcgcttggctgaa420

aatgattcttctttttatcgtctgcggcggcgttctgtttctgcttcggtatgtgattgt480

gaagctggcttacagaagagcggtaaaagaagaaataaaaaagaaatcatcttttttgtt540

tggaaagcgagggaagcgttcacagtttcgggcagctttttttataggaacattgatttg600

tattcactctgccaagttgttttgatagagtgattgtgataattttaaatgtaagcgtta660

acaaaattctccagtcttcacatcggtttgaaaggaggaagcggaagaatgaagtaagag720

ggatttttgactccgaagtaagtcttcaaaaaatcaaataaggagtgtcaagagagcgga780

taacaatttcacacaggaaacagctatgaccatgattacgaattcgagctcggtacccgg840

ggatcctctagagataccgttcgtatagcatacattatacgaagttatcttgatatggct900

ttttatatgtgttactctacatacagaaaggaggaactaaacatggccaagttgaccagt960

gccgttccggtgctcaccgcgcgcgacgtcgccggagcggtcgagttctggaccgaccgg1020

ctcgggttctcccgggacttcgtggaggacgacttcgccggtgtggtccgggacgacgtg1080

accctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtg1140

tgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttc1200

cgggacgcctccgggccggccatgaccgagatcggcgagcagccgtgggggcgggagttc1260

gccctgcgcgacccggccggcaactgcgtgcacttcgtggccgaggagcaggactgaata1320

acttcgtatagcatacattatacgaacggtaaatctgataggtggtatgttttcgcttga1380

acttttaaatacagccattgaacatacggttgatttaataactgacaaacatcaccctct1440

tgctaaagcggccaaggacgccgccgccggggctgtttgcgttcttgccgtgatttcgtg1500

taccattggtttacttatttttttgccaaggctgtaatggctgaaaattcttacatttat1560

tttacatttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatataaagtgat1620

agcggtaccattataggtaagagaggaatgtacacatgtactatttaaaaaacacaaact1680

tttggatgttcggtttattctttttcttttacttttttatcatgggagcctacttcccgt1740

ttttcccgatttggctacatgacatcaaccatatcagcaaaagtgatacgggtattattt1800

ttgccgctatttctctgttctcgctattattccaaccgctgtttggtctgctttctgaca1860

aactcgggctgcgcaaatacctgctgtggattattaccggcatgttagtgatgtttgcgc1920

cgttctttatttttatcttcgggccactgttacaatacaacattttagtaggatcgattg1980

ttggtggtatttatctaggcttttgttttaacgccggtgcgccagcagtagaggcattta2040

ttgagaaagtcagccgtcgcagtaatttcgaatttggtcgcgcgcggatgtttggctgtg2100

ttggctgggcgctgtgtgcctcgattgtcggcatcatgttcaccatcaataatcagtttg2160

ttttctggctgggctctggctgtgcactcatcctcgccgttttactctttttcgccaaaa2220

cggatgcgccctcttctgccacggttgccaatgcggtaggtgccaaccattcggcattta2280

gccttaagctggcactggaactgttcagacagccaaaactgtggtttttgtcactgtatg2340

ttattggcgtttcctgcacctacgatgtttttgaccaacagtttgctaatttctttactt2400

cgttctttgctaccggtgaacagggtacgcgggtatttggctacgtaacgacaatgggcg2460

aattacttaacgcctcgattatgttctttgcgccactgatcattaatcgcatcggtggga2520

aaaacgccctgctgctggctggcactattatgtctgtacgtattattggctcatcgttcg2580

ccacctcagcgctggaagtggttattctgaaaacgctgcatatgtttgaagtaccgttcc2640

tgctggtgggctgctttaaatatattaccagccagtttgaagtgcgtttttcagcgacga2700

tttatctggtctgtttctgcttctttaagcaactggcgatgatttttatgtctgtactgg2760

cgggcaatatgtatgaaagcatcggtttccagggcgcttatctggtgctgggtctggtgg2820

cgctgggcttcaccttaatttccgtgttcacgcttagcggccccggcccgctttccctgc2880

tgcgtcgtcaggtgaatgaagtcgcttaagggcaaggctagacgggacttaccgaaagaa2940

accatcaatgatggtttcttttttgttcataaatcagacaaaacttttctcttgcaaaag3000

tttgtgaagtgttgcacaatataaatgtgaaatacttcacaaacaaaaagacatcaaaga3060

gaaacataccctggaaggatgattaatgatgaacaaacatgtaaataaagtagctttaat3120

cggagcgggttttgttggaagcagttatgcatttgcgttaattaaccaaggaatcacaga3180

tgagcttgtggtcattgatgtaaataaagaaaaagcaatgggcgatgtgatggatttaaa3240

ccacggaaaggcgtttgcgccacaaccggtcaaaacatcttacggaacatatgaagactg3300

caaggatgctgatattgtctgcatttgcgccggagcaaaccaaaaacctggtgagacacg3360

ccttgaattagtagaaaagaacttgaagattttcaaaggcatcgttagtgaagtcatggc3420

gagcggatttgacggcattttcttagtcgcgacaaatccggttgatatcctgacttacgc3480

aacatggaaattcagcggcctgccaaaagagcgggtgattggaagcggcacaacacttga3540

ttctgcgagattccgtttcatgctgagcgaatactttggcgcagcgcctcaaaacgtaca3600

cgcgcatattatcggagagcacggcgacacagagcttcctgtttggagccacgcgaatgt3660

cggcggtgtgccggtcagtgaactcgttgagaaaaacgatgcgtacaaacaagaggagct3720

ggaccaaattgtagatgatgtgaaaaacgcagcttaccatatcattgagaaaaaaggcgc3780

gacttattatggggttgcgatgagtcttgct3811

<210>14

<211>30

<212>dna

<213>人工序列(未知)

<400>14

catacataggaagcaggcattgttcataac30

<210>15

<211>47

<212>dna

<213>人工序列(未知)

<400>15

atacgggatcaaatccgatgaaagagaaaaaatcgtacactgagctc47

<210>16

<211>42

<212>dna

<213>人工序列(未知)

<400>16

aagggggaaatgggatccatgagtcagaaaacagacgcacct42

<210>17

<211>29

<212>dna

<213>人工序列(未知)

<400>17

actacctcagttgaaggctataatccaag29

<210>18

<211>52

<212>dna

<213>人工序列(未知)

<400>18

tttctctttcatcggatttgatcccgtataccgttcgtatagcatacattat52

<210>19

<211>42

<212>dna

<213>人工序列(未知)

<400>19

gactcatggatcccatttccccctttgatttttagatatcac42

<210>20

<211>34

<212>dna

<213>人工序列(未知)

<400>20

gacccgcgtccggtttttgataagccgaagaaag34

<210>21

<211>35

<212>dna

<213>人工序列(未知)

<400>21

agtaaattcagttgcgcccggaccaatttggacac35

<210>22

<211>4568

<212>dna

<213>人工序列(未知)

<400>22

gacccgcgtccggtttttgataagccgaagaaagcgaaaaaattagagggcaaaaccgct60

attattactggaggagacagcgggattggacgcgctgtctcggtgttattcgcaaaagaa120

ggggctaatgtggtcattgtgtatttgaatgaacatcaggacgccgaggaaacaaagcag180

tatgtagaaaaggaaggggtaaaatgcctgctgattgcaggagatgtcggggatgaagcg240

ttttgcaatgatgtggtcgggcaggcaagccaagtgtttccatccattgatatattggtt300

aacaatgcagctgagcagcatgtccagcccagcattgaaaaaatcaccagccaccagctg360

atcagaaccttccaaacaaacattttttccatgttttacttaacaaaggcagtgctgcct420

catttaaaaaaagggagctctattattaataccgcttcaattaccgcctataaaggcaat480

aaaacgctgatcgattattcagcgacaaaaggcgcgatcgttacgtttacaaggtccctt540

tcccagtcgcttgttcagcagggcatacgggtcaatgcagtggcaccgggtcccatttgg600

acaccgcttatcccggccagctttgccgcaaaagacgtggaagtgtttggttcagacgtg660

ccgatggaacgcccggggcagccggtcgaagtggcgccaagctatctataccttgccagc720

gacgattccacatacgtcacagggcagacgattcacgtcaatggcggaacaattgtgaac780

ggataataaaaagccaatccttgaaggattggcttattcgctctgcttgagcgctgcata840

ttctttagagagcgtaagaaacgcatctttatctttttcgtccaaggctttgttaatttg900

gtccgtcagcatggcaagccgacggttatacaaggattcatcgagcaccatttgaatgta960

aatatccgtcattgtgacatcgaattcttttgttttttgcgtgttgcgggacttcatgag1020

ctcagtgtacgattttttctctttcatcggatttgatcccgtataccgttcgtatagcat1080

acattatacgaagttatgccatagtgactggcgatgctgtcggaatggacgacggcaata1140

gttacccttattatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaa1200

aaaaacacaaaagaccacattttttaatgtggtcttttattcttcaactaaagcacccat1260

tagttcaacaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttac1320

agtaatattgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctc1380

ctaaattcactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgat1440

ttagacaattggaagagaaaagagatatttaatcattatttgaaccaacaaacgactttt1500

agtataaccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatat1560

aaattttaccctgcatttattttcttagtgacaagggtgataaactcaaatacagctttt1620

agaactggttacaatagcgacggagagttaggttattgggataagttagagccactttat1680

acaatttttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgac1740

ttcaaagagttttatgatttatacctttctgatgtagagaaatataatggttcggggaaa1800

ttgtttcccaaaacacctatacctgaaaatgctttttctctttctattattccatggact1860

tcatttactgggtttaacttaaatatcaataataatagtaattaccttctacccattatt1920

acagcaggaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggta1980

catcattctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattg2040

tcagataggcctaatgactggcttttataatatgagataatgccgactgtactttttaca2100

gtcggttttctaaaacgatacattaataggtacgaaaaagcaactttttttgcgcttaaa2160

accagtcataccaataaataacttcgtatagcatacattatacgaacggtattcagaacg2220

ctcggttgccgccgggcgttttttatgcagcaatggcaagaacgtcccggggagctccta2280

acttataggggtaacacttaaaaaagaatcaataacgatagaaaccgctcctaaagcagg2340

tgcattttttcctaacgaagaaggcaatagttcacatttattgtctaaatgagaatggac2400

tctagaagaaacttcgtttttaatcgtatttaaaacaatgggatgagattcaattatatg2460

atttctcaagataacagcttctatatcaaatgtattaaggatattggttaatccaattcc2520

gatataaaagccaaagttttgaagtgcatttaacatttctacatcatttttatttgcgcg2580

ttccacaatctcttttcgagaaatattcttttcttctttagagagcgaagccagtaacgc2640

tttttcagaagcatataattcccaacagcctcgatttccacagctgcatttgggtccatt2700

aaaatctatcgtcatatgacccatttccccagaaaaaccctgaacacctttatacaattc2760

gttgttaataacaagtccagttccaattccgatattaatactgatgtaaacgatgttttc2820

atagttttttgtcataccaaatactttttcaccgtatgctcctgcattagcttcattttc2880

aacaaaaaccggaacattaaactcactctcaattaaaaactgcaaatctttgatattcca2940

atttaagttaggcatgaaaataatttgctgatgacgatctacaaggcctggaacacaaat3000

tcctattccgactagaccataaggggactcaggcatatgggttacaaaaccatgaataag3060

tgcaaataaaatctcttttacttcactagcggaagaactagacaagtcagaagtcttctc3120

gagaataatatttccttctaagtcggttagaattccgttaagatagtcgactcctatatc3180

aataccaatcgagtagcctgcattcttattaaaaacaagcattacaggtcttctgccgcc3240

tctagattgccctgccccaatttcaaaaataaaatctttttcaagcagtgtatttacttg3300

agaggagacagtagacttgtttaatcctgtaatctcagagagagttgccctggagacagg3360

ggagttcttcaaaatttcatctaatattaatttttgattcattttttttactaaagcttg3420

atctgcaatttgaataataaccactcctttgtttatccaccgaactaagttggtgttttt3480

tgaagcttgaattagatatttaaaagtatcatatctaatattataactaaattttctaaa3540

aaaaacattgaaataaacatttattttgtatatgatgagataaagttagtttattggata3600

aacaaactaactcaattaagatagttgatggataaacttgttcacttaaatcaaaggggg3660

aaatgacaaatggtccaaactagtgatatctaaaaatcaaagggggaaatgggatccatg3720

agtcagaaaacagacgcacctttagaatcgtatgaagtgaacggcgcaacaattgccgtg3780

ctgccagaagaaatagacggcaaaatctgttccaaaattattgaaaaagattgcgtgttt3840

tatgtaaacatgaagccgctgcaaattgtcgacagaagctgccgattttttggatcaagc3900

tatgcgggaagaaaagcaggaacttatgaagtgacaaaaatttcacacaagccgccgatc3960

atggtggacccttcgaaccaaatctttttattccctacactttcttcgacaagaccccaa4020

tgcggctggatttcccatgtgcatgtaaaagaattcaaagcgactgaattcgacgatacg4080

gaagtgacgttttccaatgggaaaacgatggagctgccgatctcttataattcgttcgag4140

aaccaggtataccgaacagcgtggctcagaaccaaattccaagacagaatcgaccaccgc4200

gtgccgaaaagacaggaatttatgctgtacccgaaagaagagcggacgaagatgatttat4260

gattttattttgcgtgagctcggggaacggtattagaaaaataggaaggagctgaccgaa4320

cagggcagctcctttcataaagctatgccgtcggaattcctccggtgatgccatatacct4380

gagatgtcacatagcttgaattctctgaagctagaaatacataaacatcggccaattcca4440

ctggctgcccagcgcgatttaatggagctggcggcgtgccttgtccgaatttcgggatgt4500

tttccgtcggctgtccgccggaaatttgcagcggtgtccaaattggtccgggcgcaactg4560

aatttact4568

<210>23

<211>25

<212>dna

<213>人工序列(未知)

<400>23

gggacaaggaatagtaagccggcaa25

<210>24

<211>56

<212>dna

<213>人工序列(未知)

<400>24

tcctgtgtgaaattgttatccgctcctacatactctctgtagcagaggtagcttga56

<210>25

<211>59

<212>dna

<213>人工序列(未知)

<400>25

tgaagcccgcctaatgagcgggcttttttctgataagaactgcaaaagctgcggattat59

<210>26

<211>33

<212>dna

<213>人工序列(未知)

<400>26

ccaccctatagataaatttttcggctgccatat33

<210>27

<211>36

<212>dna

<213>人工序列(未知)

<400>27

agagtatgtaggagcggataacaatttcacacagga36

<210>28

<211>57

<212>dna

<213>人工序列(未知)

<400>28

aaagaaatgacatgattttgcatgtgtacattcctctcttacctataatggtaccgc57

<210>29

<211>55

<212>dna

<213>人工序列(未知)

<400>29

aggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagcag55

<210>30

<211>55

<212>dna

<213>人工序列(未知)

<400>30

cccgctcattaggcgggcttcatcattactgaaacggaacgataaactgttcgcg55

<210>31

<211>24

<212>dna

<213>人工序列(未知)

<400>31

cgtatccgatttgggtgagtgtga24

<210>32

<211>28

<212>dna

<213>人工序列(未知)

<400>32

tgatgtcaatctaatgcctccttactgg28

<210>33

<211>4313

<212>dna

<213>人工序列(未知)

<400>33

cgtatccgatttgggtgagtgtgacaatcaatccagcaccgcttggagaaagtccaattg60

ccttactaattaatccgaccaagggttgtgcatagtaaagattagcaacaatgataccgc120

atgcagttgcgagaagaagggttaaactgggtgaaatcctctgacctgcttgtttgtttc180

cattagccatatgaatgaactcctttttaagtggttttttgagtgtaatcatgacttctc240

acaagtgattttataaactgaacgttcagtttcgtcgattaataaaaataatatactaaa300

cgttcagttttgtaaatggatttctgttttctttttttattcaatttaagtatactgaac360

gtatagtttatattgcgggaaggaatgatttatgtgcagagtaaacgagggcggccgcgt420

gatgaaggaacgcataaggcgattctctctgcagcctatgacctattgctggaaaaaggc480

ttcgatgcggtgacagtcgataaaattgccgagcgtgcgaaagtgagtaaagcaacgatt540

tataaatggtggtctaacaaggctgccgttatcatggacagctttctttcgaccgcgacg600

gacaggctgcctgtgcctgatacagggtcgtcagtacaagatatagtaacccacgccacg660

aatttagctaggtttttgacaagccgggaaggaaccgttattaaggaattaataggtgca720

gggcagttggatgaaaaattggcagaagaatatcgcacgcgatttttccagcctcgccgc780

ctccaagcgaaaggccttctagaaaagggaattcagaaaggtgaattgagagagaatctt840

gatattgaagtaagcatagatctcatttacggaccaattttctatcgtctgcttataaca900

ggggatgaggtgaatgattcctatgtgcgtgatttggtgatgaatgcgtttaagggagtt960

caagctacctctgctacagagagtatgtaggagcggataacaatttcacacaggaaacag1020

ctatgaccatgattacgaattcgagctcggtacccggggatcctctagagattgtaccgt1080

tcgtatagcatacattatacgaagttatcgattttcgttcgtgaatacatgttataataa1140

ctataactaataacgtaacgtgactggcaagagatatttttaaaacaatgaataggttta1200

cacttactttagttttatggaaatgaaagatcatatcatatataatctagaataaaatta1260

actaaaataattattatctagataaaaaatttagaagccaatgaaatctataaataaact1320

aaattaagtttatttaattaacaactatggatataaaataggtactaatcaaaatagtga1380

ggaggatatatttgaatacatacgaacaagttaataaagtgaaaaaaatacttcggaaac1440

atttaaaaaataaccttattggtacttacatgtttggatcaggagttgagagtggactaa1500

aaccaaatagtgatcttgactttttagtcgtcgtatctgaaccattgacagatcaaagta1560

aagaaatacttatacaaaaaattagacctatttcaaaaaaaataggagataaaagcaact1620

tacgatatattgaattaacaattattattcagcaagaaatggtaccgtggaatcatcctc1680

ccaaacaagaatttatttatggagaatggttacaagagctttatgaacaaggatacattc1740

ctcagaaggaattaaattcagatttaaccataatgctttaccaagcaaaacgaaaaaata1800

aaagaatatacggaaattatgacttagaggaattactacctgatattccattttctgatg1860

tgagaagagccattatggattcgtcagaggaattaatagataattatcaggatgatgaaa1920

ccaactctatattaactttatgccgtatgattttaactatggacacgggtaaaatcatac1980

caaaagatattgcgggaaatgcagtggctgaatcttctccattagaacatagggagagaa2040

ttttgttagcagttcgtagttatcttggagagaatattgaatggactaatgaaaatgtaa2100

atttaactataaactatttaaataacagattaaaaaaattataaataacttcgtatagca2160

tacattatacgaacggtagaatcgtcgactgataggtggtatgttttcgcttgaactttt2220

aaatacagccattgaacatacggttgatttaataactgacaaacatcaccctcttgctaa2280

agcggccaaggacgccgccgccggggctgtttgcgttcttgccgtgatttcgtgtaccat2340

tggtttacttatttttttgccaaggctgtaatggctgaaaattcttacatttattttaca2400

tttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatataaagtgatagcggt2460

accattataggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcag2520

cagcggaaagacgcgctcatattgccgatacatttggcagacatggaatcccgtttcaat2580

ttttcgatgcgcttatgccgtcagaacgcttagaacaggcaatggcggaattagttccgg2640

gcctgtcagctcatccgtatcttagcggagtggaaaaagcatgctttatgagccatgcgg2700

tcttatggaaacaagctcttgatgaaggcctgccgtacatcacagtttttgaagatgatg2760

tgctgcttggcgaaggagccgaaaaatttctggcagaagatgcgtggcttcaggaaagat2820

ttgatccggatacagcatttatcgtgcgcttagaaacaatgtttatgcatgtcctgacat2880

caccgagcggcgttgccgattattgtggaagagcatttccgttactggaatctgaacatt2940

ggggcacagcgggatacatcatctcaagaaaagctatgagatttttcctggatagatttg3000

ctgcccttccgccggaaggcttacatccggttgatctgatgatgttttctgatttctttg3060

atcgcgaaggaatgccggtgtgccaactgaatccggctctttgtgcccaggaacttcatt3120

acgccaaatttcatgatcaaaacagcgcactgggatctcttatcgaacatgatagacttc3180

tgaaccgcaaacaacagagacgcgatagcccggcgaacacatttaaacatagattaattc3240

gcgctctgacaaaaatctctagagaacgcgaaaaaagacgccaaagacgcgaacagttta3300

tcgttccgtttcagtaatgatgaagcccgcctaatgagcgggcttttttctgataagaac3360

tgcaaaagctgcggattatatgacaaattggttccatcgtacagtgaaatataagatttt3420

tgttgctgagcggaatcatcgagggaaaagttctcttatcggatgtcgaaaaagaagtca3480

ttcaatcgtgaatacataacatcatattattacattccttttcatcgattggaacaaggg3540

ttctatacttcaatggaataccgcaaagttggagtatttaaacaacaggggaaaatcgat3600

ggccattatgtaggtgtcatggcaatggaaaaaatacttgcaatgtaaacaggcctctaa3660

agagacctgttttttaatatccagatgattgtctagtttcagcttgggctctatgaccac3720

tcccataactgaaatagtctaatatttataccagaagggattgtttatataaataaaagt3780

ttatataatgacgggggcgctaaggaaacgcggggagcgactcaggttttcgtttcaaag3840

ttttgatccttaagatatacggagcgctacatatggaaacgtaaaggggaatgtataatc3900

atttggcgtgtatataacgatttgcttatatattgatttgggaaggaggagctcatatga3960

cgattgatgttgcggcgatgactcgttgtttgaaaacacttagtgatcaaactaggctta4020

tcatgatgagattatttcttgaacaagaatattgtgtctgtcaattggttgatatgttcg4080

aaatgagccagcccgccataagccagcatttgcggaaattaaaaaacgcaggttttgtga4140

atgaggacagaagaggccaatggcgttattattcaataaatggttcctgtcctgagtttg4200

atacattgcaattgatcttacatcaaattgatcaagaggatgaattgttaaaccatatca4260

aacaaaagaaaactcaagcgtgttgccagtaaggaggcattagattgacatca4313

<210>34

<211>59

<212>dna

<213>人工序列(未知)

<400>34

tgaagcccgcctaatgagcgggcttttttaagcttggcgtaatcatggtcatagctgtt59

<210>35

<211>56

<212>dna

<213>人工序列(未知)

<400>35

ctgtgtgaaattgttatccgctcccagcctttcttattaaaaccactttgtcagcc56

<210>36

<211>7781

<212>dna

<213>人工序列(未知)

<400>36

tgaagcccgcctaatgagcgggcttttttaagcttggcgtaatcatggtcatagctgttt60

cctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaag120

tgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactg180

cccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcg240

gggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgc300

tcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatcc360

acagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccagg420

aaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcat480

cacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccag540

gcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccgga600

tacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtagg660

tatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgtt720

cagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacac780

gacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggc840

ggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtattt900

ggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatcc960

ggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgc1020

agaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtgg1080

aacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctag1140

atccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttgg1200

tctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgt1260

tcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttacca1320

tctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatca1380

gcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcc1440

tccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagt1500

ttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatg1560

gcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgc1620

aaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtg1680

ttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaaga1740

tgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcga1800

ccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaacttta1860

aaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctg1920

ttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttact1980

ttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaata2040

agggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatt2100

tatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaa2160

ataggggttccgcgcacatttccccgaaaagtgccacctgacgtctaagaaaccattatt2220

atcatgacattaacctataaaaataggcgtatcacgaggccctttcgtctcgcgcgtttc2280

ggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctg2340

taagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgt2400

cggggctggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcgg2460

tgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgccattcgccattc2520

aggctgcgcaactgttgggaagggcgatcggtgcgggcctcttcgctattacgccagctg2580

gcgaaagggggatgtgctgcaaggcgattaagttgggtaacgccagggttttcccagtca2640

cgacgttgtaaaacgacggccagtgaattccttaaggaacgtacagacggcttaaaagcc2700

tttaaaaacgtttttaaggggtttgtagacaaggtaaaggataaaacagcacaattccaa2760

gaaaaacacgatttagaacctaaaaagaacgaatttgaactaactcataaccgagaggta2820

aaaaaagaacgaagtcgagatcagggaatgagtttataaaataaaaaaagcacctgaaaa2880

ggtgtctttttttgatggttttgaacttgttctttcttatcttgatacatatagaaataa2940

cgtcatttttattttagttgctgaaaggtgcgttgaagtgttggtatgtatgtgttttaa3000

agtattgaaaacccttaaaattggttgcacagaaaaaccccatctgttaaagttataagt3060

gactaaacaaataactaaatagatgggggtttcttttaatattatgtgtcctaatagtag3120

catttattcagatgaaaaatcaagggttttagtggacaagacaaaaagtggaaaagtgag3180

accatggagagaaaagaaaatcgctaatgttgattactttgaacttctgcatattcttga3240

atttaaaaaggctgaaagagtaaaagattgtgctgaaatattagagtataaacaaaatcg3300

tgaaacaggcgaaagaaagttgtatcgagtgtggttttgtaaatccaggctttgtccaat3360

gtgcaactggaggagagcaatgaaacatggcattcagtcacaaaaggttgttgctgaagt3420

tattaaacaaaagccaacagttcgttggttgtttctcacattaacagttaaaaatgttta3480

tgatggcgaagaattaaataagagtttgtcagatatggctcaaggatttcgccgaatgat3540

gcaatataaaaaaattaataaaaatcttgttggttttatgcgtgcaacggaagtgacaat3600

aaataataaagataattcttataatcagcacatgcatgtattggtatgtgtggaaccaac3660

ttattttaagaatacagaaaactacgtgaatcaaaaacaatggattcaattttggaaaaa3720

ggcaatgaaattagactatgatccaaatgtaaaagttcaaatgattcgaccgaaaaataa3780

atataaatcggatatacaatcggcaattgacgaaactgcaaaatatcctgtaaaggatac3840

ggattttatgaccgatgatgaagaaaagaatttgaaacgtttgtctgatttggaggaagg3900

tttacaccgtaaaaggttaatctcctatggtggtttgttaaaagaaatacataaaaaatt3960

aaaccttgatgacacagaagaaggcgatttgattcatacagatgatgacgaaaaagccga4020

tgaagatggattttctattattgcaatgtggaattgggaacggaaaaattattttattaa4080

agagtagttcaacaaacgggccagtttgttgaagattagatgctataattgttattaaaa4140

ggattgaaggatgcttaggaagacgagttattaatagctgaataagaacggtgctctcca4200

aatattcttatttagaaaagcaaatctaaaattatctgaaaagggaatgagaatagtgaa4260

tggaccaataataatgactagagaagaaagaatgaagattgttcatgaaattaaggaacg4320

aatattggataaatatggggatgatgttaaggctattggtgtttatggctctcttggtcg4380

tcagactgatgggccctattcggatattgagatgatgtgtgtcatgtcaacagaggaagc4440

agagttcagccatgaatggacaaccggtgagtggaaggtggaagtgaattttgatagcga4500

agagattctactagattatgcatctcaggtggaatcagattggccgcttacacatggtca4560

atttttctctattttgccgatttatgattcaggtggatacttagagaaagtgtatcaaac4620

tgctaaatcggtagaagcccaaacgttccacgatgcgatttgtgcccttatcgtagaaga4680

gctgtttgaatatgcaggcaaatggcgtaatattcgtgtgcaaggaccgacaacatttct4740

accatccttgactgtacaggtagcaatggcaggtgccatgttgattggtctgcatcatcg4800

catctgttatacgacgagcgcttcggtcttaactgaagcagttaagcaatcagatcttcc4860

ttcaggttatgaccatctgtgccagttcgtaatgtctggtcaactttccgactctgagaa4920

acttctggaatcgctagagaatttctggaatgggattcaggagtggacagaacgacacgg4980

atatatagtggatgtgtcaaaacgcataccattttgaacgatgacctctaataattgtta5040

atcatgttggttacgtatttattaacttctcctagtattagtaattatcatggctgtcat5100

ggcgcattaacggaataaagggtgtgcttaaatcgggccattttgcgtaataagaaaaag5160

gattaattatgagcgaattgaattaataataaggtaatagatttacattagaaaatgaaa5220

ggggattttatgcgtgagaatgttacagtctatcccggcattgccagtcggggatattaa5280

aaagagtataggtttttattgggataaagtaggtttcactttggttcaccatgaagatgg5340

attcgcagttctaatgtgtaatgaggttcggattcatctatgggaggcaagtgatgaagg5400

ctggcgcctcgtagtaatgattcaccggtttgtacaggtgcggagtcgtttattgctggt5460

actgctagttgccgcattgaagtagagggaattgatgaattatatcaacatattaagcct5520

ttgggcattttgcaccccaatacatcattaaaagatcagtggtgggatgaacgagacttt5580

gcagtaattgatcccgacaacaatttgattagcttttttcaacaaataaaaagctaaaat5640

ctattattaatctgttcagcaatcgggcgcgattgctgaataaaagatacgagagacctc5700

tcttgtatcttttttattttgagtggttttgtccgttacactagaaaaccgaaagacaat5760

aaaaattttattcttgctgagtctggctttcggtaagctagacaaaacggacaaaataaa5820

aattggcaagggtttaaaggtggagattttttgagtgatcttctcaaaaaatactacctg5880

tcccttgctgatttttaaacgagcacgagagcaaaacccccctttgctgaggtggcagag5940

ggcaggtttttttgtttcttttttctcgtaaaaaaaagaaaggtcttaaaggttttatgg6000

ttttggtcggcactgccgcgcctcgcagagcacacactttatgaatataaagtatagtgt6060

gttatactttacttggaagtggttgccggaaagagcgaaaatgcctcacatttgtgccac6120

ctaaaaaggagcgatttacatatgagttatgcagtttgtagaatgcaaaaagtgaaatca6180

gctggactaaaaggcatgcaatttcataatcaaagagagcgaaaaagtagaacgaatgat6240

gatattgaccatgagcgaacacgtgaaaattatgatttgaaaaatgataaaaatattgat6300

tacaacgaacgtgtcaaagaaattattgaatcacaaaaaacaggtacaagaaaaacgagg6360

aaagatgctgttcttgtaaatgagttgctagtaacatctgaccgagatttttttgagcaa6420

ctggatcctgataggtggtatgttttcgcttgaacttttaaatacagccattgaacatac6480

ggttgatttaataactgacaaacatcaccctcttgctaaagcggccaaggacgccgccgc6540

cggggctgtttgcgttcttgccgtgatttcgtgtaccattggtttacttatttttttgcc6600

aaggctgtaatggctgaaaattcttacatttattttacatttttagaaatgggcgtgaaa6660

aaaagcgcgcgattatgtaaaatataaagtgatagcggtaccattataggtaagagagga6720

atgtacacatgccgtctgaagcttttagacgccatagagcctatcgcgaaaataaacttc6780

aaccgttagtctctgttttaatctgcgcttataacgtggaaaaatattttgcccaatcac6840

tggcagcggttgtgaatcagacatggagaaacctggatattcttatcgtcgatgatggct6900

caacagatggaacacttgctattgcccaaagatttcaagaacaggatggcagaattcgca6960

tcttagcacagccgcgcaattctggccttattccgtcattaaacatcggattagatgaac7020

tggcgaaaagcggcggaggcggagaatatatcgctagaacagatgccgatgatattgctg7080

ccccggattggattgaaaaaatcgttggcgaaatggaaaaagatagatcaatcatcgcaa7140

tgggagcgtggcttgaagtgttaagcgaagaaaaagatggcaatagactggctcgccatc7200

atgaacatggaaaaatctggaaaaaaccgacaagacatgaagatattgccgatttctttc7260

cgtttggcaatccgattcataataacacaatgatcatgagacgcagcgtcattgatggcg7320

gacttagatataacacagaacgcgattgggcggaagattaccagttttggtacgatgttt7380

ctaaactgggaagacttgcatattatccggaagcgctggtgaaatatcgccttcatgcta7440

atcaagtctcaagcaaatatagcatcagacagcatgaaattgcacaaggcatccagaaaa7500

cagcgcgcaacgattttcttcaaagcatgggatttaaaacaagatttgattctctggaat7560

accgccagatcaaagcagttgcgtatgaactgcttgaaaaacatctgccggaagaagatt7620

ttgaaagagcaagacgctttctttaccaatgttttaaacgcacagatacattaccggctg7680

gcgcctggctggattttgcagcggatggaagaatgagacgcttatttacactgcgccagt7740

actttggaatcctgcatagactgctgaaaaaccgctaatga7781

<210>37

<211>33

<212>dna

<213>人工序列(未知)

<400>37

ggctttcgttgtcttgtgttcaagaaatttcca33

<210>38

<211>56

<212>dna

<213>人工序列(未知)

<400>38

ctgtgtgaaattgttatccgctcccagcctttcttattaaaaccactttgtcagcc56

<210>39

<211>54

<212>dna

<213>人工序列(未知)

<400>39

aagcccgcctaatgagcgggcttttttatcataaagatccagcctttttgcgct54

<210>40

<211>32

<212>dna

<213>人工序列(未知)

<400>40

ctaaagcccaagtcacaatatattgatcgcct32

<210>41

<211>51

<212>dna

<213>人工序列(未知)

<400>41

gttttaataagaaaggctgggagcggataacaatttcacacaggaaacagc51

<210>42

<211>57

<212>dna

<213>人工序列(未知)

<400>42

aaagaaatgacatgattttgcatgtgtacattcctctcttacctataatggtaccgc57

<210>43

<211>55

<212>dna

<213>人工序列(未知)

<400>43

aggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagcag55

<210>44

<211>55

<212>dna

<213>人工序列(未知)

<400>44

cccgctcattaggcgggcttcatcattactgaaacggaacgataaactgttcgcg55

<210>45

<211>31

<212>dna

<213>人工序列(未知)

<400>45

aacctccttgtcttctttccagtcttatctc31

<210>46

<211>29

<212>dna

<213>人工序列(未知)

<400>46

atcacttctctttctttcacgctcatcct29

<210>47

<211>3627

<212>dna

<213>人工序列(未知)

<400>47

aacctccttgtcttctttccagtcttatctctaggatgaaattggaggaagaataacgtg60

catgtcattacaacacaagtactttttattttttgttttttattgctgattcactcgata120

gaaaccttagcctatgcgacaaggctttccggagctcgcgttggatttattgcgtccgcg180

ctttctctgtttaatgtcatggtcatcgtatccagaatgtcgaatatggtgcagcagccc240

tttactgggcatttaattgatgatgctggaaaaaacgcactggcgattgtaggggagcag300

ttccgctttttaattttcggatcgacagtcggcaccattttgggcattatcctgctcccg360

tcttttgtcgctcttttttcacgggcgattattcacttggcgggcggcggcggctccgtt420

tttcaagtattccgaaagggattctcgaaacaaggattcaaaaatgccctttcctatttg480

cgtctgccgtccatttcatatgtaaaaggatttcatatgcgcttgattccgaagcgtttg540

tttgtcatcaacatgctgatcacatcgatttatacgattggtgtgctttcggctttatac600

gcaggccttttggcgccggagcgcagcacgacagccgtcatggcttcgggtttgatcaac660

ggaattgcaacgatgctgctggctatttttgttgatcctaaggtatccgttcttgctgat720

gatgtggcaaaaggaaaacgaagctatatctatttaaaatggacctctgtcacaatggtc780

acatcaagggtggcgggcacactcctcgcccagctcatgtttattcccggggcctactat840

atcgcgtggctgacaaagtggttttaataagaaaggctgggagcggataacaatttcaca900

caggaaacagctatgaccatgattacgaattcgagctcggtacccggggatcctctagag960

ataccgttcgtatagcatacattatacgaagttatcttgatatggctttttatatgtgtt1020

actctacatacagaaaggaggaactaaacatggccaagttgaccagtgccgttccggtgc1080

tcaccgcgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctccc1140

gggacttcgtggaggacgacttcgccggtgtggtccgggacgacgtgaccctgttcatca1200

gcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtgtgggtgcgcggcc1260

tggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccg1320

ggccggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacc1380

cggccggcaactgcgtgcacttcgtggccgaggagcaggactgaataacttcgtatagca1440

tacattatacgaacggtaaatcgtcgactgataggtggtatgttttcgcttgaactttta1500

aatacagccattgaacatacggttgatttaataactgacaaacatcaccctcttgctaaa1560

gcggccaaggacgccgccgccggggctgtttgcgttcttgccgtgatttcgtgtaccatt1620

ggtttacttatttttttgccaaggctgtaatggctgaaaattcttacatttattttacat1680

ttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatataaagtgatagcggta1740

ccattataggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagc1800

agcggaaagacgcgctcatattgccgatacatttggcagacatggaatcccgtttcaatt1860

tttcgatgcgcttatgccgtcagaacgcttagaacaggcaatggcggaattagttccggg1920

cctgtcagctcatccgtatcttagcggagtggaaaaagcatgctttatgagccatgcggt1980

cttatggaaacaagctcttgatgaaggcctgccgtacatcacagtttttgaagatgatgt2040

gctgcttggcgaaggagccgaaaaatttctggcagaagatgcgtggcttcaggaaagatt2100

tgatccggatacagcatttatcgtgcgcttagaaacaatgtttatgcatgtcctgacatc2160

accgagcggcgttgccgattattgtggaagagcatttccgttactggaatctgaacattg2220

gggcacagcgggatacatcatctcaagaaaagctatgagatttttcctggatagatttgc2280

tgcccttccgccggaaggcttacatccggttgatctgatgatgttttctgatttctttga2340

tcgcgaaggaatgccggtgtgccaactgaatccggctctttgtgcccaggaacttcatta2400

cgccaaatttcatgatcaaaacagcgcactgggatctcttatcgaacatgatagacttct2460

gaaccgcaaacaacagagacgcgatagcccggcgaacacatttaaacatagattaattcg2520

cgctctgacaaaaatctctagagaacgcgaaaaaagacgccaaagacgcgaacagtttat2580

cgttccgtttcagtaatgatgaagcccgcctaatgagcgggcttttttatcataaagatc2640

cagcctttttgcgcttccttatttatagttccttaatccgcggtcgtacaaaaaatttcc2700

gaatgggacgaaagcggcgatgaagccggcagctgaccatttcagcggccatttgacaga2760

gaaggttgcataagccaagacaagcaaatacaaaatgaacaacccgccgtgaaccgaacc2820

gacaattgtcaccgcaagcggaaggcctgcccaatatttaagcggcatagcgatgaacaa2880

caggattaagagtgacattccttcaataaaacccatcgtgcgaagtcttccgatcggcgt2940

gtgcagcataaatcgccctccttgtggacacgttttcattttatactataaacaatccgg3000

gggggcatatgacagctttcaaaaaatgttcggaaaacattcatttttacatgccttttc3060

tagggaactgtacttgtcatttacaaaaatacccgagataatgtgtacaaaatcaaaaaa3120

gaaggatgttgaaatgaaacttgaccagattgatctgaatatcattgaggagctgaagaa3180

ggacagccgtttgtcgatgagggaattaggcagaaaaattaagctgtcgcctccatctgt3240

aacagaacgggtaagacagcttgaatcgtttggcatcatcaagcaatacacgctggaggt3300

cgaccagaaaaaactggggcttcccgtttcctgcattgtggaagcaaccgttaaaaacgc3360

ggattatgagcggttcaaaagctatattcaaacattgccgaatattgaattttgctaccg3420

gattgcgggtgcagcctgctatatgctgaaaatcaatgccgaaagcctcgaagcggtaga3480

agatttcattaacaaaacatcgccctacgcgcaaaccgtcactcacgtcattttctcaga3540

aattgacacgaaaaacgggcgcggttagagagtgccgcgcgaagtctgttataataacag3600

gatgagcgtgaaagaaagagaagtgat3627

<210>48

<211>35

<212>dna

<213>人工序列(未知)

<400>48

catccagcgtaaaacgttcacgggaataatctagg35

<210>49

<211>50

<212>dna

<213>人工序列(未知)

<400>49

ttatccgctctggcgcggacttgtttgtttatatccattctaaatgaagg50

<210>50

<211>59

<212>dna

<213>人工序列(未知)

<400>50

gcccgcctaatgagcgggcttttttttaagacgtggactcgttttcagcctgaaatttt59

<210>51

<211>29

<212>dna

<213>人工序列(未知)

<400>51

catcccagcagctgatcaggatgaattct29

<210>52

<211>49

<212>dna

<213>人工序列(未知)

<400>52

acaagtccgcgccagagcggataacaatttcacacaggaaacagctatg49

<210>53

<211>56

<212>dna

<213>人工序列(未知)

<400>53

agaaatgacatgattttgcatgtgtacattcctctcttacctataatggtaccgct56

<210>54

<211>59

<212>dna

<213>人工序列(未知)

<400>54

ataggtaagagaggaatgtacacatgcaaaatcatgtcatttctttagcatcagcagcg59

<210>55

<211>57

<212>dna

<213>人工序列(未知)

<400>55

gcccgctcattaggcgggcttcatcattactgaaacggaacgataaactgttcgcgt57

<210>56

<211>26

<212>dna

<213>人工序列(未知)

<400>56

gccaggaagccacagcacatcatagc26

<210>57

<211>35

<212>dna

<213>人工序列(未知)

<400>57

ttcgttcattcgttcccgtttacagaacaaatagc35

<210>58

<211>4150

<212>dna

<213>人工序列(未知)

<400>58

gccaggaagccacagcacatcatagccctgcatgcgtttcattcttgttacaatgtcttg60

cagcgttgtgtcccatgcgtggccaaggtgaagtctccctgttacgtttggcggcgggat120

gacaacagagtatggctcttttgtctggtcactgcccgcttcaaaaaatttgccttttag180

ccaaaaatcatatcggtctttttcaaccgctgccggatcatatttcgtcggcattgtttg240

ttcattcgtttccatcataaccctccagcatgtttaaaaatggctgagaacgaaaaaaga300

gctcttcatccctgtaaaaggacgaaaagctctttcgcggtaccaccttttttcatgaac360

tcaatcagttcatgcacttataacggataacgggtgtttcccgaatttctctactcttct420

ttcaagaaattatgctcccgggcgaccttccaatcagacaacataagaagcctttcagca480

aacggcttctctctctgcatgcgccttttttgtactcttcccgttctaagcaggtgtatt540

tatcagctcattgtatacattatttatgttctttattatagtaaaagtgtttttgattcg600

tcaatcatcttttccaaaaacaccgctttctatacagatgggcaggcacctattgcccaa660

gtttacatatgatgcttgtaaatgcgtagaggaggatgcaccgtgaagaaacgtttcagc720

tcttattcgctgccgccatgggtaaggcaaattcggcttgtatccgcacaagtgattatt780

cccattacgatttttcaaggaatcagaaccattttctttccgacaacctttgatgttttg840

ctgctcgcaatcctaatctttttagcttgcgcccttcatttagaatggatataaacaaac900

aagtccgcgccagagcggataacaatttcacacaggaaacagctatgaccatgattacga960

attcgagctcggtacccggggatcctctagagattgtaccgttcgtatagcatacattat1020

acgaagttatgccatagtgactggcgatgctgtcggaatggacgacggcaatagttaccc1080

ttattatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaaca1140

caaaagaccacattttttaatgtggtcttttattcttcaactaaagcacccattagttca1200

acaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaata1260

ttgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaatt1320

cactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagaca1380

attggaagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataa1440

ccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatataaatttt1500

accctgcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactg1560

gttacaatagcgacggagagttaggttattgggataagttagagccactttatacaattt1620

ttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaag1680

agttttatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttc1740

ccaaaacacctatacctgaaaatgctttttctctttctattattccatggacttcattta1800

ctgggtttaacttaaatatcaataataatagtaattaccttctacccattattacagcag1860

gaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcatt1920

ctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagata1980

ggcctaatgactggcttttataatatgagataatgccgactgtactttttacagtcggtt2040

ttctaacgatacattaataggtacgaaaaagcaactttttttgcgcttaaaaccagtcat2100

accaataaataacttcgtatagcatacattatacgaacggtatgataggtggtatgtttt2160

cgcttgaacttttaaatacagccattgaacatacggttgatttaataactgacaaacatc2220

accctcttgctaaagcggccaaggacgccgccgccggggctgtttgcgttcttgccgtga2280

tttcgtgtaccattggtttacttatttttttgccaaggctgtaatggctgaaaattctta2340

catttattttacatttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatata2400

aagtgatagcggtaccattataggtaagagaggaatgtacacatgcaaaatcatgtcatt2460

tctttagcatcagcagcggaaagacgcgctcatattgccgatacatttggcagacatgga2520

atcccgtttcaatttttcgatgcgcttatgccgtcagaacgcttagaacaggcaatggcg2580

gaattagttccgggcctgtcagctcatccgtatcttagcggagtggaaaaagcatgcttt2640

atgagccatgcggtcttatggaaacaagctcttgatgaaggcctgccgtacatcacagtt2700

tttgaagatgatgtgctgcttggcgaaggagccgaaaaatttctggcagaagatgcgtgg2760

cttcaggaaagatttgatccggatacagcatttatcgtgcgcttagaaacaatgtttatg2820

catgtcctgacatcaccgagcggcgttgccgattattgtggaagagcatttccgttactg2880

gaatctgaacattggggcacagcgggatacatcatctcaagaaaagctatgagatttttc2940

ctggatagatttgctgcccttccgccggaaggcttacatccggttgatctgatgatgttt3000

tctgatttctttgatcgcgaaggaatgccggtgtgccaactgaatccggctctttgtgcc3060

caggaacttcattacgccaaatttcatgatcaaaacagcgcactgggatctcttatcgaa3120

catgatagacttctgaaccgcaaacaacagagacgcgatagcccggcgaacacatttaaa3180

catagattaattcgcgctctgacaaaaatctctagagaacgcgaaaaaagacgccaaaga3240

cgcgaacagtttatcgttccgtttcagtaatgatgaagcccgcctaatgagcgggctttt3300

ttttaagacgtggactcgttttcagcctgaaattttttctcttgttcaatctgatgaatt3360

ttcatcacaaacggctctatgtttttcatttgccaataagcctttaataattgtgtacat3420

tcctctttttcatgctgtcttttcccagtttgataaccgttcagcaccttgtacatggct3480

gccggatatgccatatagctgagaaaaaggctgacttccgcttcacgcaaagggaatgat3540

ttcgtatatccataaaaccattcgacgcattccggacaagctttcggaaagcctctgaac3600

atttttgtataaaagccgagcaggtcgttttgcggcggaccgacagatgctctttcgaag3660

tttgtaaaatatccggtcccggcatcattatacagaaaatggtgtatagacaggctgccg3720

tggttcattacaacccttgaaacgtccttttcctttgcggactcataccagtcctccagc3780

cgttccagcgcaaagttcaccgcggaaatggtttcagaaaaataggttatggcctggagt3840

tcaaagggagaaaggtaccattccttctccgctctatccacaaactgctcatagaaaatt3900

ttatcttgttcccacttttttttcgtttgcccgtaataacgttcaatctcatcacgtcta3960

acctttagttcctgcgcggttctctcgtgaagacgggctgtttctctaaataaataagca4020

tgtttttgatcccgttcttcctcttggtcaaattggagccaaggcattaaatagtagata4080

tctccgtattgtatgcctgctgagaaaaactcaccgctatttgttctgtaaacgggaacg4140

aatgaacgaa4150

<210>59

<211>29

<212>dna

<213>人工序列(未知)

<400>59

cgaaggaatgccggtgtgccaactgaatc29

<210>60

<211>30

<212>dna

<213>人工序列(未知)

<400>60

cgacctccagcgtgtattgcttgatgatgc30

<210>61

<211>30

<212>dna

<213>人工序列(未知)

<400>61

gggcacagcgggatacatcatctcaagaaa30

<210>62

<211>29

<212>dna

<213>人工序列(未知)

<400>62

tgaaccacggcagcctgtctatacaccat29

<210>63

<211>31

<212>dna

<213>人工序列(未知)

<400>63

gagggtacaaagatgtgaactatacgcttgc31

<210>64

<211>57

<212>dna

<213>人工序列(未知)

<400>64

gaaattgttatccgctcaaggccttaagccttataccttttttaattttaagtccga57

<210>65

<211>61

<212>dna

<213>人工序列(未知)

<400>65

gcccgcctaatgagcgggcttttttatacctttttgattaatcattccattgatacgtcc60

a61

<210>66

<211>32

<212>dna

<213>人工序列(未知)

<400>66

catgaaagcgatcataaaagaagatgtgcaag32

<210>67

<211>50

<212>dna

<213>人工序列(未知)

<400>67

ggtataaggcttaaggccttgagcggataacaatttcacacaggaaacag50

<210>68

<211>53

<212>dna

<213>人工序列(未知)

<400>68

caaagcgtacatgcgtcatgtgtacattcctctcttacctataatggtaccgc53

<210>69

<211>44

<212>dna

<213>人工序列(未知)

<400>69

agaggaatgtacacatgacgcatgtacgctttgactactcaaaa44

<210>70

<211>58

<212>dna

<213>人工序列(未知)

<400>70

gcccgctcattaggcgggcttcatcattaatcttccagacgtttttcaagctctgctt58

<210>71

<211>31

<212>dna

<213>人工序列(未知)

<400>71

gctgtttgcgttcttgccgtgatttcgtgta31

<210>72

<211>37

<212>dna

<213>人工序列(未知)

<400>72

gataaatgcccttctcatctttcccttcgctttctcc37

<210>73

<211>3900

<212>dna

<213>人工序列(未知)

<400>73

gttcaacatggtatgaaacaatcgaggagactgatcccaattccatttatgcctcaacag60

accctgttgtcattcacaatttaaagagccagaatcttgcctttaactataaaataaacc120

gtatttttatagatgatgaggacgtatacttgcctatactgaaagaatgggcctttgaag180

tcacacaggatcaagagcatttaaaaactcctatccactatattattcgggagtttgtca240

gagtgagagatttatatgtttcttatgtgaaagaatttgtccatctcaatcagaatactg300

tgaaaagcgaagaagccgaagacctgtatcacgcccttataaaagcgtttgatcttgtga360

ttcatatttttatagaagaaatgtacaaaaacacaagccttcagcttcaggcccaaaaag420

atatgattactgaattgagcgctccggtcatcgtgctgtttcacagcgtcggactgctgc480

ctttaatcggagatattgatacagtccgcgccaagctgattatggaaaacacactgcatc540

aatgtgcgaaaaaaaaggtgacacaactgtatattgatttgtcaggagtagctgtgattg600

atacgatggttgcccatcagctgttcagtctgattgaggcgcttcgtttaatcggtgttt660

cttcaactttgtctggaatccgtccggaaatcgcgcaaacagcggttcagctaggtttgt720

ctttcgaaggtatttccttgagatccactctcgcttcagctattgcatcggacttaaaat780

taaaaaaggtataaggcttaaggccttgagcggataacaatttcacacaggaaacagcta840

tgaccatgattacgaattcgagctcggtacccggggatcctctagagataccgttcgtat900

agcatacattatacgaagttatcttgatatggctttttatatgtgttactctacatacag960

aaaggaggaactaaacatggccaagttgaccagtgccgttccggtgctcaccgcgcgcga1020

cgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtgga1080

ggacgacttcgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtccagga1140

ccaggtggtgccggacaacaccctggcctgggtgtgggtgcgcggcctggacgagctgta1200

cgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccggccatgac1260

cgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactg1320

cgtgcacttcgtggccgaggagcaggactgaataacttcgtatagcatacattatacgaa1380

cggtaaatcgtcgactgataggtggtatgttttcgcttgaacttttaaatacagccattg1440

aacatacggttgatttaataactgacaaacatcaccctcttgctaaagcggccaaggacg1500

ccgccgccggggctgtttgcgttcttgccgtgatttcgtgtaccattggtttacttattt1560

ttttgccaaggctgtaatggctgaaaattcttacatttattttacatttttagaaatggg1620

cgtgaaaaaaagcgcgcgattatgtaaaatataaagtgatagcggtaccattataggtaa1680

gagaggaatgtacacatgacgcatgtacgctttgactactcaaaagcgttgactttcttc1740

aacgaacatgaacttacatacctgcgggactttgtaaaaacagcacaccataatatccat1800

gagaaaacaggcgcgggcagcgattttctaggctgggtggacctccctgaacattatgat1860

aaagaagaattcgcgcgcatcaaaaaaagcgcggaaaaaatcaaatctgactctgatgtc1920

ttgcttgttgtcggcatcggcggttcttatcttggagcgcgggcagcgattgaagcgctg1980

aatcacgcgttttataacactttgccaaaagcaaaacgcggcaatccgcaagtcattttt2040

atcgggaacaacatcagttcatcttatatgagagacgtcatggatcttcttgaagatgtt2100

gacttctctattaatgtgatttctaaatcaggtacgacaactgaacctgcaatcgctttc2160

cgtattttccgcaagcttcttgaagagaaatacggtaaagaagaagcgaaagcgcggatt2220

tatgcaacaactgataaagagcgcggcgcattaaaaacgctttctaacgaagaaggcttt2280

gaatcattcgtaattcctgacgatgtcggcggccgttattcagttttaacagctgtaggt2340

ctcttgccgattgctgtcagcggcgtcaacattgacgacatgatgaaaggcgccctggat2400

gcgagcaaagattttgcaacatctgaactggaagataacccagcataccaatatgcggtt2460

gttcgcaatgtcctttataataagggcaaaacaattgaaatgctcatcaactacgaaccg2520

gcgcttcaatactttgcggaatggtggaagcagctgttcggagaaagcgaagggaaagat2580

gagaagggcatttatccttcttcagcgaactattcaacagaccttcattctttaggccag2640

tatgtacaagaaggccgcagagatttattcgaaacggtcctgaacgtagagaagcctaaa2700

catgaactgacaattgaggaagcggataacgatcttgacggcttgaactatttagccggt2760

aaaactgttgatttcgttaacaaaaaagcattccaaggtacaatgcttgcccatacagac2820

ggaaatgttccgaacttaatcgttaacattcctgagctgaatgcatatacttttggatac2880

cttgtatatttcttcgaaaaagcctgcgcgatgagcggttacctccttggcgtcaatccg2940

tttgaccagcctggtgtagaagcgtataaagtcaatatgtttgcgttactcggcaaacct3000

ggctttgaagagaaaaaagcagagcttgaaaaacgtctggaagattaatgatgaagcccg3060

cctaatgagcgggcttttttatacctttttgattaatcattccattgatacgtccaatat3120

atttcttcttcaaaccatgttgccataacaggttcattatttttcaatttctcttcaagc3180

tctctgagcatgccttctgtttgtgtgcggtgagctcttaacgcattcattttaatatca3240

gctacttctttgatatcaagtacgacatcagcttcgccgagaacctcttctctgttgcgg3300

gtaatggccatgcagattgtacgcggacggtcttctttctttttgcggtataatgcacgg3360

ataaccgcttccccgcatgcatcatggtcagggtgcacaccgtgtccgggataaaatgtg3420

acaattaaacttggttttacatcatctatgatctcttccatgatatcagctaaatactca3480

tcgtcttcaaattcgagcgttttgtcacggagtccaagcatccgaaggtcattgatatcc3540

atttctttgcaagcgttgatgagctcttgttttcttagcagcggcaacgtttcccgattc3600

gcgaaaaatggatcgcccatatttctgcccatttcacctaaagtggcgcaggcatacgta3660

acgggaatgtctttttttctgtttaaggcaatgaggcctgccactccatatgattcatca3720

tcagggtgaggcaggattactagtacatgttctttcattattattcacccttttctttaa3780

actgtgaacggtttttcgctgatttgcagagaaatagccagtttaccgcccggcaggtgt3840

cccgccattaataaacggttttcttcatctaccgtgtattcagtcaagccttcagcataa3900

<210>74

<211>31

<212>dna

<213>人工序列(未知)

<400>74

gagggtacaaagatgtgaactatacgcttgc31

<210>75

<211>57

<212>dna

<213>人工序列(未知)

<400>75

gaaattgttatccgctcaaggccttaagccttataccttttttaattttaagtccga57

<210>76

<211>63

<212>dna

<213>人工序列(未知)

<400>76

taaagcccgcctaatgagcgggcttttttacgagtatgtcatacgttcgtctgtactgca60

aaa63

<210>77

<211>25

<212>dna

<213>人工序列(未知)

<400>77

ccattttaaagatcgcagctgcggc25

<210>78

<211>47

<212>dna

<213>人工序列(未知)

<400>78

agcagtttcatgtgtaggagcggataacaatttcacacaggaaacag47

<210>79

<211>52

<212>dna

<213>人工序列(未知)

<400>79

acctacgattccacacatgtgtacattcctctcttacctataatggtaccgc52

<210>80

<211>46

<212>dna

<213>人工序列(未知)

<400>80

agaggaatgtacacatgtgtggaatcgtaggttatatcggtcagct46

<210>81

<211>53

<212>dna

<213>人工序列(未知)

<400>81

cccgctcattaggcgggctttactccacagtaacactcttcgcaaggttacga53

<210>82

<211>32

<212>dna

<213>人工序列(未知)

<400>82

tgtttcgtttgactcgtcgttgtttgcttcct32

<210>83

<211>33

<212>dna

<213>人工序列(未知)

<400>83

ctaggcgatgtagttgtccatgtcttccacaag33

<210>84

<211>5335

<212>dna

<213>人工序列(未知)

<400>84

tgtttcgtttgactcgtcgttgtttgcttcctgctttaccactgtattttctgtctctgt60

tggtacagcttgcttcaccggctctttatttttccctgtggccagaaagatcataatcgc120

tgtgaaaacagccgcagaagccgctaaaataattgctttcttatgctgattcaaccaatt180

catcgtgcatgttccccgctttcgtcatattttattttcataacacatatggtttagagg240

ataatagctcaaaggaggggaaaccattgaagataggctttatcggcacaggaaatatgg300

gtacgatccttatcgaatcatttattgaatcaaaagcggccgatccctcaaacatgacaa360

tcacaaaccgcacaattgaaaaagcgttgcacataaaaaaccgctacaacagcataaacg420

ttaccgaaagtctcgaaaagctcgtttctgaaaacgagatgatttttatttgtgttaagc480

cgcttgatatatatcctttactggcaagagcgttaccttatttaagaaaagatcatattc540

tcatttcaattacaagccccgttcaaaccgaacagctcgaacaatatgtgccgtgtcaag600

tggcgcgagtgatcccaagtattacgaacagagcgctggcaggtgtttctctcgtcactt660

tcggcacaagctgcggcgaatctgcaaaagcgaaaatcaatgagctgatgcagcatatct720

ctcatccgctgcaaattgaaagtgacatcacgagggttgcatcagacatcgtaagctgcg780

gccccgcttttatgagctatctgattcagcggtttatagatgcagctgtttcggaaacat840

cagtgtctaaacaggacgcgattttgatgtgtaaggaaatgcttgtcggaatgggaaaac900

tgttagaaaccgaattgtataccctgcctgcattacaggaaaaggtttgcgttaaaggcg960

gtgtaacaggagaaggcattaaagcgctggaaagcggtgtgcaggatatgtttcaccggg1020

tatttcaaaatactcatatgaagtacgaggaggatatttccgcagtgaaaaagcagtttc1080

atgtgtaggagcggataacaatttcacacaggaaacagctatgaccatgattacgaattc1140

gagctcggtacccggggatcctctagagattgtaccgttcgtatagcatacattatacga1200

agttatcgattttcgttcgtgaatacatgttataataactataactaataacgtaacgtg1260

actggcaagagatatttttaaaacaatgaataggtttacacttactttagttttatggaa1320

atgaaagatcatatcatatataatctagaataaaattaactaaaataattattatctaga1380

taaaaaatttagaagccaatgaaatctataaataaactaaattaagtttatttaattaac1440

aactatggatataaaataggtactaatcaaaatagtgaggaggatatatttgaatacata1500

cgaacaagttaataaagtgaaaaaaatacttcggaaacatttaaaaaataaccttattgg1560

tacttacatgtttggatcaggagttgagagtggactaaaaccaaatagtgatcttgactt1620

tttagtcgtcgtatctgaaccattgacagatcaaagtaaagaaatacttatacaaaaaat1680

tagacctatttcaaaaaaaataggagataaaagcaacttacgatatattgaattaacaat1740

tattattcagcaagaaatggtaccgtggaatcatcctcccaaacaagaatttatttatgg1800

agaatggttacaagagctttatgaacaaggatacattcctcagaaggaattaaattcaga1860

tttaaccataatgctttaccaagcaaaacgaaaaaataaaagaatatacggaaattatga1920

cttagaggaattactacctgatattccattttctgatgtgagaagagccattatggattc1980

gtcagaggaattaatagataattatcaggatgatgaaaccaactctatattaactttatg2040

ccgtatgattttaactatggacacgggtaaaatcataccaaaagatattgcgggaaatgc2100

agtggctgaatcttctccattagaacatagggagagaattttgttagcagttcgtagtta2160

tcttggagagaatattgaatggactaatgaaaatgtaaatttaactataaactatttaaa2220

taacagattaaaaaaattataaataacttcgtatagcatacattatacgaacggtagaat2280

cgtcgactgataggtggtatgttttcgcttgaacttttaaatacagccattgaacatacg2340

gttgatttaataactgacaaacatcaccctcttgctaaagcggccaaggacgccgccgcc2400

ggggctgtttgcgttcttgccgtgatttcgtgtaccattggtttacttatttttttgcca2460

aggctgtaatggctgaaaattcttacatttattttacatttttagaaatgggcgtgaaaa2520

aaagcgcgcgattatgtaaaatataaagtgatagcggtaccattataggtaagagaggaa2580

tgtacacatgtgtggaatcgtaggttatatcggtcagcttgatgcgaaggaaattttatt2640

aaaagggttagagaagcttgagtatcgcggttatgactctgctggtattgctgttgccaa2700

cgaacagggaatccatgtgttcaaagaaaaaggacgcattgcagatcttcgtgaagttgt2760

ggatgccaatgtagaagcgaaagccggaattgggcatactcgctgggcgacacacggcga2820

accaagctatctgaacgctcacccgcatcaaagcgcactgggccgctttacacttgttca2880

caacggcgtgatcgagaactatgttcagctgaagcaagagtatttgcaagatgtagagct2940

caaaagtgacaccgatacagaagtagtcgttcaagtaatcgagcaattcgtcaatggagg3000

acttgagacagaagaagcgttccgcaaaacacttacactgttaaaaggctcttatgcaat3060

tgctttattcgataacgacaacagagaaacgatttttgtagcgaaaaacaaaagccctct3120

attagtaggtcttggagatacattcaacgtcgtagcatctgatgcgatggcgatgcttca3180

agtaaccaacgaatacgtagagctgatggataaagaaatggttatcgtcactgatgacca3240

agttgtcatcaaaaaccttgatggtgacgtgattacacgtgcgtcttatattgctgagct3300

tgatgccagtgatatcgaaaaaggcacgtaccctcactacatgttgaaagaaacggatga3360

gcagcctgttgttatgcgcaaaatcatccaaacgtatcaagatgaaaacggcaagctgtc3420

tgtgcctggcgatatcgctgccgctgtagcggaagcggaccgcatctatatcattggctg3480

cggaacaagctaccatgcaggacttgtcggtaaacaatatattgaaatgtgggcaaacgt3540

gccggttgaagtgcatgtagcgagtgaattctcctacaacatgccgcttctgtctaagaa3600

accgctcttcattttcctttctcaaagcggagaaacagcagacagccgcgcggtactcgt3660

tcaagtcaaagcgctcggacacaaagccctgacaatcacaaacgtacctggatcaacgct3720

ttctcgtgaagctgactatacattgctgcttcatgcaggccctgagatcgctgttgcgtc3780

aacgaaagcatacactgcacaaatcgcagttctggcggttcttgcttctgtggctgctga3840

caaaaatggcatcaatatcggatttgacctcgtcaaagaactcggtatcgctgcaaacgc3900

aatggaagctctatgcgaccagaaagacgaaatggaaatgatcgctcgtgaatacctgac3960

tgtatccagaaatgctttcttcatcggacgcggccttgactacttcgtatgtgtcgaagg4020

cgcactgaagctgaaagagatttcttacatccaggcagaaggttttgccggcggtgagct4080

aaagcacggaacgattgccttgatcgaacaaggaacaccagtattcgcactggcaactca4140

agagcatgtaaacctaagcatccgcggaaacgtcaaagaagttgctgctcgcggagcaaa4200

cacatgcatcatctcactgaaaggcctagacgatgcggatgacagattcgtattgccgga4260

agtaaacccagcgcttgctccgttggtatctgttgttccattgcagctgatcgcttacta4320

tgctgcactgcatcgcggctgtgatgtggataaacctcgtaaccttgcgaagagtgttac4380

tgtggagtaaagcccgcctaatgagcgggcttttttacgagtatgtcatacgttcgtctg4440

tactgcaaaagcataaaaaagcattttaaaaaaccatcgttttaggaaacgatggttttt4500

gatttctgcgccttgaaaaagaggcgttctgattgtgctgatggctcggtatccgtaaaa4560

tcagctgtcacgcgatggagctgaaagccgcagttcttaagcatttcttcgtattcttcc4620

accgggaacgttctttgctcgtgtgtctcatcaaaacgatcatacgcttctccgttccac4680

acaaaaaacgacatatcatgaatgacagaaagctcgtcacttccggcaaagctctgccaa4740

atatagctgatatcttcgtcctggtcagcaaatgtgctatccggaaagacttccgcaatt4800

ttaaaagatgaatgaacatcaaacagcaggatgccttcaggctttaatacccgaaaaaca4860

cttttaaaggtttcgatcacatcattttttgtttttaaataatttaatgagtcacagcat4920

atcacaacagcgtcaaattgaccgtcaaagccggtaatctctctcatatcctgctgaaga4980

aataaaataggctggctactcgataccttttgctgggcgaaagaaagcatctcttcgctg5040

agatcgatgcctgtaacctcgaagcctttttcggcaagccggatggagatttcccccgtg5100

ccgcatgccagatcgagaatacggcctttttccgggagagatgcttcaatccattttgtc5160

cattgatcgtaaggcgcgtgtgacattagctcgtcatacacgcttgcaaagccttgataa5220

atcattgattcattccaaagtcaagatctgcgagcggagcgtctccccacagcttttcaa5280

ggttgtagtagcttctttcatccttgtggaagacatggacaactacatcgcctag5335

<210>85

<211>28

<212>dna

<213>人工序列(未知)

<400>85

ggcaacggttcatctttttctcagaacg28

<210>86

<211>54

<212>dna

<213>人工序列(未知)

<400>86

cctgtgtgaaattgttatccgctcttattccacgagataaagagtttgctggct54

<210>87

<211>61

<212>dna

<213>人工序列(未知)

<400>87

gcccgcctaatgagcgggcttttttatacctttttgattaatcattccattgatacgtcc60

a61

<210>88

<211>32

<212>dna

<213>人工序列(未知)

<400>88

catgaaagcgatcataaaagaagatgtgcaag32

<210>89

<211>48

<212>dna

<213>人工序列(未知)

<400>89

ctttatctcgtggaataagagcggataacaatttcacacaggaaacag48

<210>90

<211>53

<212>dna

<213>人工序列(未知)

<400>90

caaagcgtacatgcgtcatgtgtacattcctctcttacctataatggtaccgc53

<210>91

<211>54

<212>dna

<213>人工序列(未知)

<400>91

aggtaagagaggaatgtacacatgaaaaaagtacgtaaagccataattccagca54

<210>92

<211>63

<212>dna

<213>人工序列(未知)

<400>92

gcccgctcattaggcgggctttagatttcttctttgtttagtaaaccttccataaatgga60

acg63

<210>93

<211>27

<212>dna

<213>人工序列(未知)

<400>93

ttcttctattatcggtgttcagcaggt27

<210>94

<211>25

<212>dna

<213>人工序列(未知)

<400>94

ggaggcgtacagagatgaaggttat25

<210>95

<211>4509

<212>dna

<213>人工序列(未知)

<400>95

tggcaacggacctggaaatggagggcctggcggcggaggcggaaaaagcgtgaatatgtt60

cggcaccggtgatcctggaccgcttcgtctcttccaatcagcactttccggccaaatcag120

ctggatgcttccattctcattgatcggattactgggcgcaatcatcagctggtaccgtga180

tcgccgcggacatgctgctaaaatgaaagaaacgctattctgggcggcttggcttgttcc240

cgtggctggtttcttcagcatcgcaggattcttccaccagtattatttgattatgctggc300

accgccgattgccgccctttctggtatcggatggtatacgatgcaccgcttatacaaaaa360

taataaagactggtccagctatcttctgccggcagccgtcttgatcacagccgtgtttca420

ggtgtatattctgagcgcttatacaagccaaatcggcagtgtatggatgtatgtcctggg480

gctcttaggactgggcatcacccttgcacttctgatgcttaaacgcagccatccgttcag540

taagctgctgaccatcatcagcttgtgcgttctgcttctcacaccagtttactggtcagc600

aacgccgcttctatacggtggcaacagcgtccttcctgaatccggcccgcagctcaaagg660

ctcagcaaacggcgggaacatgttctcttcagaggtcgacagcgggctcctgtcctatct720

gagaaaacacaataccggcgaagagtatttatttgctactttgacaaccgtaacagctgc780

gccatacatcatttacgaaaatgaatctgttatggcaatgggcgggtttaacggcacgga840

tcccattctgactgtgtcggaactgaaaaaactgatcaaagaaggcaaagtgaaatactt900

ccttctgtcagaaaacaactcaggcaacagcgaacttgtttcctggattaagaaaaacgg960

aaaagaaataacatctgacgagtacagcggttcctccagcagcacaaacagcgtgcaggg1020

aatgcgaagaggacctggcggagaaagccagcaaactctttatctcgtggaataagagcg1080

gataacaatttcacacaggaaacagctatgaccatgattacgaattcgagctcggtaccc1140

ggggatcctctagagattgtaccgttcgtatagcatacattatacgaagttatcgatttt1200

cgttcgtgaatacatgttataataactataactaataacgtaacgtgactggcaagagat1260

atttttaaaacaatgaataggtttacacttactttagttttatggaaatgaaagatcata1320

tcatatataatctagaataaaattaactaaaataattattatctagataaaaaatttaga1380

agccaatgaaatctataaataaactaaattaagtttatttaattaacaactatggatata1440

aaataggtactaatcaaaatagtgaggaggatatatttgaatacatacgaacaagttaat1500

aaagtgaaaaaaatacttcggaaacatttaaaaaataaccttattggtacttacatgttt1560

ggatcaggagttgagagtggactaaaaccaaatagtgatcttgactttttagtcgtcgta1620

tctgaaccattgacagatcaaagtaaagaaatacttatacaaaaaattagacctatttca1680

aaaaaaataggagataaaagcaacttacgatatattgaattaacaattattattcagcaa1740

gaaatggtaccgtggaatcatcctcccaaacaagaatttatttatggagaatggttacaa1800

gagctttatgaacaaggatacattcctcagaaggaattaaattcagatttaaccataatg1860

ctttaccaagcaaaacgaaaaaataaaagaatatacggaaattatgacttagaggaatta1920

ctacctgatattccattttctgatgtgagaagagccattatggattcgtcagaggaatta1980

atagataattatcaggatgatgaaaccaactctatattaactttatgccgtatgatttta2040

actatggacacgggtaaaatcataccaaaagatattgcgggaaatgcagtggctgaatct2100

tctccattagaacatagggagagaattttgttagcagttcgtagttatcttggagagaat2160

attgaatggactaatgaaaatgtaaatttaactataaactatttaaataacagattaaaa2220

aaattataaataacttcgtatagcatacattatacgaacggtagaatcgtcgactgatag2280

gtggtatgttttcgcttgaacttttaaatacagccattgaacatacggttgatttaataa2340

ctgacaaacatcaccctcttgctaaagcggccaaggacgccgccgccggggctgtttgcg2400

ttcttgccgtgatttcgtgtaccattggtttacttatttttttgccaaggctgtaatggc2460

tgaaaattcttacatttattttacatttttagaaatgggcgtgaaaaaaagcgcgcgatt2520

atgtaaaatataaagtgatagcggtaccattataggtaagagaggaatgtacacatgaaa2580

aaagtacgtaaagccataattccagcagcaggcttaggaacacgttttcttccggctacg2640

aaagcaatgccgaaagaaatgcttcctatcgttgataaacctaccattcaatacataatt2700

gaagaagctgttgaagccggtattgaagatattattatcgtaacaggaaaaagcaagcgt2760

gcgattgaggatcattttgattactctcctgagcttgaaagaaacctagaagaaaaagga2820

aaaactgagctgcttgaaaaagtgaaaaaggcttctaacctggctgacattcactatatc2880

cgccaaaaagaacctaaaggtctcggacatgctgtctggtgcgcacgcaactttatcggc2940

gatgagccgtttgcggtactgcttggtgacgatattgttcaggctgaaactccagggttg3000

cgccaattaatggatgaatatgaaaaaacactttcttctattatcggtgttcagcaggtg3060

cccgaagaagaaacacaccgctacggcattattgacccgctgacaagtgaaggccgccgt3120

tatcaggtgaaaaacttcgttgaaaaaccgcctaaaggcacagcaccttctaatcttgcc3180

atcttaggccgttacgtattcacgcctgagatcttcatgtatttagaagagcagcaggtt3240

ggcgccggcggagaaattcagctcacagacgccattcaaaagctgaatgaaattcaaaga3300

gtgtttgcttacgattttgaaggcaagcgttatgatgttggtgaaaagctcggctttatc3360

acaacaactcttgaatttgcgatgcaggataaagagcttcgcgatcagctcgttccattt3420

atggaaggtttactaaacaaagaagaaatctaaagcccgcctaatgagcgggcttttttt3480

taagcttcttcttttacatgtacctttaacacagcctgcacttcaggatgcagcttcaca3540

ggcacgtttgtatatcctaatgcacgaatgccgtccggcagttcaagcttccgcttatct3600

actttaatgttatggtctttttggagctgttcagtgatttgcttgcttgtcactgaacca3660

aataaacggccgccctcacctgattttgcgctaagctcaacagtcagtttttctaatgtc3720

tctttcaagctcttagcctgctcaagctcagcaatggcttcttttttctctttctgcttt3780

tgtccgtttaacgcgctgatgtttgacgcgttcgcttcaaccgcgaggccttttttaatt3840

agaaagttatgcgcataaccgtctgctacgtttttgacttcgcctttttttccttttcct3900

tttacatcttgtaagaaaataaccttcatctctgtacgcctccctcaaaatactcatcta3960

tggcgtgcttcagccgttccagcgcttcagaaactgaaatgccggacaactgagtcgccg4020

catttgttaaatgccctccgccttccagggcctccatgatgatctgaacatttacctcgc4080

caagcgaccgagcactgatacaaaccgtttgttcatcccgtctcgccaccgcaaatgaag4140

cctcaacctcactcatcgatagcagcgaatcggcagcctgcgcaatcagcacttgatcga4200

agtattcctcttcattttcaggaagagaagcaatcgcaatgttatctttatagaggaccg4260

tatgctggatcagcttcgcccgtttaatataggaatcaacggtttctttcagaaacttct4320

gcaccagcaccgtgtcggcgccttttgcccttaaataagaagccgcatcgaaagtccgcg4380

agcccgtgcggagagaaaagctctttgtatccactattataccagctaatagggctgttg4440

cttcaatcatattgatttttaagcgcttcggctgatattcaagcagctctgtcaccaatt4500

ccgctgtgg4509

<210>96

<211>33

<212>dna

<213>人工序列(未知)

<400>96

gtgaagcggcgtattattaaaggaaaggctgta33

<210>97

<211>51

<212>dna

<213>人工序列(未知)

<400>97

tgtgtgaaattgttatccgctctcacacttccactgtccagttaaacgggc51

<210>98

<211>54

<212>dna

<213>人工序列(未知)

<400>98

agcccgcctaatgagcgggcttttttaaatcgaaaaagaacctgcccggaggca54

<210>99

<211>27

<212>dna

<213>人工序列(未知)

<400>99

ccggatttacggtcgcttggctgtatt27

<210>100

<211>44

<212>dna

<213>人工序列(未知)

<400>100

cagtggaagtgtgagagcggataacaatttcacacaggaaacag44

<210>101

<211>55

<212>dna

<213>人工序列(未知)

<400>101

gccagtaacaagtattgccatgtgtacattcctctcttacctataatggtaccgc55

<210>102

<211>41

<212>dna

<213>人工序列(未知)

<400>102

aagagaggaatgtacacatggcaatacttgttactggcggt41

<210>103

<211>55

<212>dna

<213>人工序列(未知)

<400>103

cccgctcattaggcgggctttattccgcactcttatacccattcacattagaaga55

<210>104

<211>20

<212>dna

<213>人工序列(未知)

<400>104

aaagcccgacccgtcttcct20

<210>105

<211>21

<212>dna

<213>人工序列(未知)

<400>105

ttggatgtggagtgccgtctt21

<210>106

<211>4666

<212>dna

<213>人工序列(未知)

<400>106

aaagcccgacccgtcttccttggggtttggacaatattttacagattatatgtttgtgat60

ggactacgaagaggggattggatggcatcatccgagaattgcgccgtacgcaccgcttac120

gcttgatccgtcttcatctgtttttcattacggccaggctgtttttgaaggattaaaagc180

atacagaacagacgacggcagggtgctgctgttccgtccggatcaaaatatcaaacggct240

gaacagatcgtgtgagcgcatgagcatgccccctttagacgaagagctggtgcttgaggc300

attgacgcaattagttgagctggagaaagattgggttccaaaggaaaaaggaacgtcact360

gtatattcgtccttttgtcattgccacagaaccgagtctcggtgtgaaggcatccaggag420

ctatacatttatgatcgtgctttcgcctgtcggctcctattatggcgacgatcagctgaa480

gccggttagaatctatgtcgaagatgagtatgtgagggcggtcaacggaggagtcgggtt540

tgcaaaaacggctggaaactatgccgccagtcttcaggcacagcggaaagcgaatgaact600

gggctatgaccaggtactgtggctggacgccatcgaaaagaaatatgtggaagaagtagg660

gagcatgaacatctttttcgtcataaacggggaagctgtcacacctgctttaagcggaag720

cattttaagcggggttacacgtgcgtctgcgattgaattgattcgaagctggggcattcc780

ggttcgtgaagagagaatatcgattgatgaggtgtatgcggcctctgcacgcggagaatt840

gacagaggtctttggcacaggcacggcagcagtcgttacgcctgtcggtgaactcaacat900

ccatggaaaaacggtgattgtaggcgacgggcaaatcggggacctctcgaaaaagctgta960

tgaaacgataacagatattcagcttggcaaggtaaaaggcccgtttaactggacagtgga1020

agtgtgagagcggataacaatttcacacaggaaacagctatgaccatgattacgaattcg1080

agctcggtacccggggatcctctagagattgtaccgttcgtatagcatacattatacgaa1140

gttatgccatagtgactggcgatgctgtcggaatggacgacggcaatagttacccttatt1200

atcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaacacaaaa1260

gaccacattttttaatgtggtcttttattcttcaactaaagcacccattagttcaacaaa1320

cgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaatattgac1380

ttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaattcactt1440

tagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagacaattgg1500

aagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataaccaca1560

gaaattgatattagtgttttataccgaaacataaaacaagaaggatataaattttaccct1620

gcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactggttac1680

aatagcgacggagagttaggttattgggataagttagagccactttatacaatttttgat1740

ggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaagagttt1800

tatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttcccaaa1860

acacctatacctgaaaatgctttttctctttctattattccatggacttcatttactggg1920

tttaacttaaatatcaataataatagtaattaccttctacccattattacagcaggaaaa1980

ttcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcattctgtt2040

tgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagataggcct2100

aatgactggcttttataatatgagataatgccgactgtactttttacagtcggttttcta2160

acgatacattaataggtacgaaaaagcaactttttttgcgcttaaaaccagtcataccaa2220

taaataacttcgtatagcatacattatacgaacggtatgataggtggtatgttttcgctt2280

gaacttttaaatacagccattgaacatacggttgatttaataactgacaaacatcaccct2340

cttgctaaagcggccaaggacgccgccgccggggctgtttgcgttcttgccgtgatttcg2400

tgtaccattggtttacttatttttttgccaaggctgtaatggctgaaaattcttacattt2460

attttacatttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatataaagtg2520

atagcggtaccattataggtaagagaggaatgtacacatggcaatacttgttactggcgg2580

tgccggttacattggcagccacacatgtgttgaactattgaacagcggctacgagattgt2640

tgttcttgataatctgtccaacagttcagctgaagcgctgaaccgtgtcaaggagattac2700

aggaaaagatttaacgttctacgaagcggatttattggaccgggaagcggtagattccgt2760

ttttgctgaaaatgaaatcgaagctgtgattcattttgcagggttaaaagcagtcggcga2820

atctgtggcgattcccctcaaatattatcataacaatttgacaggaacgtttattttatg2880

cgaggccatggagaaatacggcgtcaagaaaatcgtattcagttcatctgcgacagtata2940

cggcgttccggaaacatcgccgattacggaagactttccattaggcgcgacaaatcctta3000

tgggcagacgaagctcatgcttgaacaaatattgcgtgatttgcatacagccgacaatga3060

gtggagcgttgcgctgcttcgttactttaacccgttcggcgcgcatccaagcggacggat3120

cggtgaagacccgaacggaatcccaaataaccttatgccgtatgtggcacaggtagcagt3180

cgggaagctcgagcaattaagcgtattcggaaatgactatccgacaaaagacgggacagg3240

cgtacgcgattatattcacgtcgttgatctcgcagaaggccacgtcaaggcgctggaaaa3300

agtattgaactctacaggagccgatgcatacaaccttggaacaggcacaggctacagcgt3360

gctggaaatggtcaaagcctttgaaaaagtgtcagggaaagaggttccataccgttttgc3420

ggaccgccgtccgggagacatcgccacatgctttgcagatcctgcgaaagccaagcgaga3480

actaggctgggaagcgaaacgcggccttgaggaaatgtgtgctgattcctggagatggca3540

gtcttctaatgtgaatgggtataagagtgcggaataaagcccgcctaatgagcgggcttt3600

tttaaatcgaaaaagaacctgcccggaggcaggttctttttattttgaatgagtcatatc3660

agcagccggctggcttttgcggttttttacataatatattgcatagcaaatggccataaa3720

tggaacgccgcaatagagtgcgattctttgattcggatcaaaggcaatgccaacgacgga3780

agccaggcataacaggaacgcagcaatcggcacgaacggatacaaaggggttctgtattt3840

caaatcagtaaccttgtttcccgcttcgatataacgttttctgaacatgaattgagaagc3900

agcgattcccatccacacgacaacgacggcaaaaccggaaatcgacacgagcacaacgta3960

tacagtgtctggcgcaaatacgctggacagcaacgacagaatgccgccgatcatgctgaa4020

caccagcgcgttaaaaggggttccttttgacgtcagctttgcaaatgtcggatgaagcgt4080

tttttcttttgataaagaccacagcatccgggaagaagcgtagagtccggaattagccgc4140

tgacaggatcgctgtcaggataacaaagttcatgatatcggctgcgtatggtacaccgac4200

tctgtcaaagacagcgacaaacgggcttttgatgacacctgcatcttgaatcggaatcag4260

ccctgacagcacaaaaattgttccgacaaagaagagcgagagacgccagaccgttgtttt4320

tatcgcttttggtatcgtcttgtcaggatcaacgctttcaccggctgcgattccgattag4380

ttccgttcccgaaaaggcaaaattgacggacagcatcgtcatcaaaatcgggacaaatcc4440

attgggaaaaagcccgccttccgccgtgaagttggacagcatcggagcggcttctccgcc4500

tttgatcgggatgatgccgaacatggcagagccgcctaaaagaataaataatacaatggc4560

taatactttgatgctggaaaaccaaaattctgattcagcgaaaaacttgaccgaaaatgc4620

gttaagcaagaagatgaacagcgcaaagacggcactccacatccaa4666

<210>107

<211>30

<212>dna

<213>人工序列(未知)

<400>107

caaattccttatgtcctgaccttgcatggc30

<210>108

<211>47

<212>dna

<213>人工序列(未知)

<400>108

tgtgtgaaattgttatccgctcagcgctcaaccctctcctgtaaaat47

<210>109

<211>39

<212>dna

<213>人工序列(未知)

<400>109

accctggcgttacggaaccctctgacaagtattttccgg39

<210>110

<211>31

<212>dna

<213>人工序列(未知)

<400>110

gccattctctttaaaacaggcggcaatacaa31

<210>111

<211>44

<212>dna

<213>人工序列(未知)

<400>111

cagtggaagtgtgagagcggataacaatttcacacaggaaacag44

<210>112

<211>55

<212>dna

<213>人工序列(未知)

<400>112

gccagtaacaagtattgccatgtgtacattcctctcttacctataatggtaccgc55

<210>113

<211>23

<212>dna

<213>人工序列(未知)

<400>113

ccgtaggttctgcgtcagttgtt23

<210>114

<211>26

<212>dna

<213>人工序列(未知)

<400>114

gatgtcggataatgctgcttgtactc26

<210>115

<211>2238

<212>dna

<213>人工序列(未知)

<400>115

ccgtaggttctgcgtcagttgttttagcagtcagccacaagcttcaggaagaagccaaaa60

agctttcgggctttgacagctcggttttgccaatcggcatacagctcagccgttttcagg120

ggaatgaggagacgaaagaggaaatcagaaaaagactcggactcccgctcgatcagcgtc180

tggctgtctatgtcggccggctggtcagagagaaaggcatttttgaactgtctgaagcga240

tcgaatcgctgcaggactctcctaaggcagtctttgtcggggatggtcccgcaaaatcga300

cgctgacgcagaaggggcatatcgtcacaggccaagtcccgaatcatcaggtaagagact360

atttacttgcggcagatctattcgtacttccttcttacagtgaggggatgccgactgttg420

tgattgaggcgctggcgctaagggttcctgttatttgtacggatgttggcggagtgtcgt480

ctttattcggaaagcaccaacatctgctgattaaaccgaaatccgctcaagccttggcgg540

aagcgatcactcggtacgaacatgagcaaatatggaagccggaagtagcagatgacttgt600

atgaaaccgtgcaggctcaatttgacgcaggaaaaaacgccaaggccttgcatcatcagt660

atcagacggtcacgaaaacgtccgtttaatcgtaagacactgcgaccattataaattgga720

agatcattttacaggagagggttgagcgctgagcggataacaatttcacacaggaaacag780

ctatgaccatgattacgaattcgagctcggtacccggggatcctctagagataccgttcg840

tatagcatacattatacgaagttatcttgatatggctttttatatgtgttactctacata900

cagaaaggaggaactaaacatggccaagttgaccagtgccgttccggtgctcaccgcgcg960

cgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgt1020

ggaggacgacttcgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtcca1080

ggaccaggtggtgccggacaacaccctggcctgggtgtgggtgcgcggcctggacgagct1140

gtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccggccat1200

gaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaa1260

ctgcgtgcacttcgtggccgaggagcaggactgaataacttcgtatagcatacattatac1320

gaacggtaaatcgtcgacctgcaggcatgcaagcttggcactggccgtcgttttacaacg1380

tcgtgactgggaaaaccctggcgttacggaaccctctgacaagtattttccgggcttgcc1440

gcttgaagaattggctaaagacttgggaagcgtcaatttataaaaagaaaaagtgttact1500

tggttgaagctgccggctgctaggctcggccgccggctcctgaaacatcaggaaaggggt1560

taaacatgtgagtatcaaacgatcagcggtccatacgctagcgttactggccgctgctat1620

atttggagtcgtcctattgctgggagccatccataaagatataggtttcatgcagatggc1680

tgctgtgcttgcggtgttggcaattggcctgtttttactgacgcttgcaaccgcatttac1740

aacaaaagaacggctctttatggctgtaatctatattttgatcgcctgcacttttttaaa1800

taatgcattttttgccattcatcttggatttttcagtttgttcctttaccggcttctgct1860

cattgcagcgggttgccttcatattttcggcatggttcggaatcgaacccatattgaaag1920

atggcatgggctgcaagtaaaagggattttactctttttcgccttttggtttatatatgg1980

ccttgtttctctgctgtgggccaagtcggtaacagttggcctcaagtatttagccttatt2040

ggcaatggggatcttcttcatttatcttattgtcatgtacgtgcaaaaaatggagcggct2100

gatgattgtttatgccatttggctggtgatgacggtatttctgatgatcattggttttta2160

caatcatatcacccaccatcatcttgcgagttccacattgtacagcggaccggagtacaa2220

gcagcattatccgacatc2238