Difference between revisions of "Team:HBUT-China/Notebook"

Line 60: Line 60:
 
                   BBa_K3126004 (promoter TEF2)
 
                   BBa_K3126004 (promoter TEF2)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px">
 
                     BBa_K3126005 (hexa-His gene)
 
                     BBa_K3126005 (hexa-His gene)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px">
 
                     BBa_K3126006 (nixA gene)
 
                     BBa_K3126006 (nixA gene)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px">
 
                     BBa_K3126007 (TgMTP1t2 gene)
 
                     BBa_K3126007 (TgMTP1t2 gene)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px">
 
                     BBa_K3126008 (kanR gene)
 
                     BBa_K3126008 (kanR gene)
 
                 </h4>
 
                 </h4>
−
                   <h4>
+
                   <h4 style="text-indent:28px">
 
                     BBa_K3126010 (terminator CYC1)
 
                     BBa_K3126010 (terminator CYC1)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px">
 
                       BBa_K3126011 (terminator TDH1)
 
                       BBa_K3126011 (terminator TDH1)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px"
 
                     BBa_K3126012 (terminator PGK1)
 
                     BBa_K3126012 (terminator PGK1)
 
                 </h4>
 
                 </h4>
−
                 <h4>
+
                 <h4 style="text-indent:28px">
 
                     BBa_K3126013 (terminator ENO2)
 
                     BBa_K3126013 (terminator ENO2)
 
                 </h4>
 
                 </h4>
Line 91: Line 91:
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word;">
 
                         <p style="word-wrap:break-word;">
−
                             GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCC</strong>CCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAA</strong>GGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC</strong>AGGCCTTACTAGTAGCGGCCG
+
                             GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCC<strong>CCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAA</strong>GGATCC<strong>TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC</strong>AGGCCTTACTAGTAGCGGCCG
 
                         </p>
 
                         </p>
 
                     </el-collapse-item>
 
                     </el-collapse-item>
Line 99: Line 99:
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
−
                             GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATA</strong>GGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACG</strong>AGGCCTTACTAGTAGCGGCCGCTGCAG
+
                             GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATA</strong>GGATCC<strong>ATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACG</strong>AGGCCTTACTAGTAGCGGCCGCTGCAG
 
                         </p>
 
                         </p>
 
                     </el-collapse-item>
 
                     </el-collapse-item>
Line 115: Line 115:
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
−
                             GAATTCGCGGCCGCTTCTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAAC</strong>GGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAAT</strong>AAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCAG
+
                             GAATTCGCGGCCGCTTCTAGAG<strong>TTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAAC</strong>GGATCC<strong>AGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAAT</strong>AAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCAG
 
                         </p>
 
                         </p>
 
                     </el-collapse-item>
 
                     </el-collapse-item>
 
                     <el-collapse-item name="15">
 
                     <el-collapse-item name="15">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <p style="font-size:16px">hexa-His</p>
+
                             <p style="font-size:16px">BBa_K3126005（hexa-His）</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
Line 128: Line 128:
 
                     <el-collapse-item name="16">
 
                     <el-collapse-item name="16">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <p style="font-size:16px">NixA</p>
+
                             <p style="font-size:16px">BBa_K3126006（nixA）</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
−
                             GGTCTCACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAA</strong>GGATCCATAAAGCAATCTTGATGAGGAGAGACC
+
                             GGTCTCACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTAT<strong>CCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAA</strong>GGATCCATAAAGCAATCTTGATGAGGAGAGACC
 
                         </p>
 
                         </p>
 
                     </el-collapse-item>
 
                     </el-collapse-item>
 
                     <el-collapse-item name="17">
 
                     <el-collapse-item name="17">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <p style="font-size:16px">TgMTP1t2</p>
+
                             <p style="font-size:16px">BBa_K3126007（TgMTP1t2）</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
−
                             GGTCTCAAACACACATAAACAAACAAAGGATCCATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAA</strong>GGATCCATTGAATTGAATTGAAATCGAGAGACC
+
                             GGTCTCAAACACACATAAACAAACAAAGGATCC<strong>ATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAA</strong>GGATCCATTGAATTGAATTGAAATCGAGAGACC
 
                         </p>
 
                         </p>
 
                     </el-collapse-item>
 
                     </el-collapse-item>
 
                     <el-collapse-item name="18">
 
                     <el-collapse-item name="18">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <p style="font-size:16px">KanR</p>
+
                             <p style="font-size:16px">BBa_K3126008（kanR）</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
Line 159: Line 159:
 
                     <el-collapse-item name="21">
 
                     <el-collapse-item name="21">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <h4>BBa_K3126001 (pSB1C3-pTEF1/tCYC1)&nbsp;&nbsp;&nbsp;</h4>
+
                             <h4> pSB1C3-pTEF1/tCYC1</h4>
−
                             <p style="font-size:16px">The sequence of BBa_K3126001</p>
+
                             <p style="font-size:16px">The sequence of pSB1C3-pTEF1/tCYC1</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word;">
 
                         <p style="word-wrap:break-word;">
Line 168: Line 168:
 
                     <el-collapse-item name="22">
 
                     <el-collapse-item name="22">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <h4>BBa_K3126002 (pSB1C3-pGPM1/tTDH1)&nbsp;&nbsp;&nbsp;</h4>
+
                             <h4>pSB1C3-pGPM1/tTDH1</h4>
−
                             <p style="font-size:16px">The sequence of BBa_K3126002</p>
+
                             <p style="font-size:16px">The sequence of pSB1C3-pGPM1/tTDH1</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">
Line 177: Line 177:
 
                     <el-collapse-item name="23">
 
                     <el-collapse-item name="23">
 
                         <template slot="title">
 
                         <template slot="title">
−
                             <h4>BBa_K3126003 (pSB1C3-pTDH3/tPGK1)&nbsp;&nbsp;&nbsp;</h4>
+
                             <h4>pSB1C3-pTDH3/tPGK1</h4>
−
                             <p style="font-size:16px">The sequence of BBa_K3126003</p>
+
                             <p style="font-size:16px">The sequence of pSB1C3-pTDH3/tPGK1</p>
 
                         </template>
 
                         </template>
 
                         <p style="word-wrap:break-word">
 
                         <p style="word-wrap:break-word">

Revision as of 10:30, 17 October 2019

2019.5

We synthesized:

BBa_K3126001 (promoter TEF1)

BBa_K3126002 (promoter GPM1)

BBa_K3126003 (promoter TDH3)

BBa_K3126004 (promoter TEF2)

BBa_K3126005 (hexa-His gene)

BBa_K3126006 (nixA gene)

BBa_K3126007 (TgMTP1t2 gene)

BBa_K3126008 (kanR gene)

BBa_K3126010 (terminator CYC1)

BBa_K3126011 (terminator TDH1)

BBa_K3126013 (terminator ENO2)

GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAAGGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAGGCCTTACTAGTAGCGGCCG

GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCCTTACTAGTAGCGGCCGCTGCAG

GAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCAGTTCGAGTTTATCATTATCAATACTGCCATTTCAAAGAATACGTAAATAATTAATAGTAGTGATTTTCCTAACTTTATTTAGTCAAAAAATTAGCCTTTTAATTCTGCTGTAACCCGTACATGCCCAAAATAGGGGGCGGGTTACACAGAATATATAACATCGTAGGTGTCTGGGTGAACAGTTTATTCCTGGCATCCACTAAATATAATGGAGCCCGCTTTTTAAGCTGGCATCCAGAAAAAAAAAGAATCCCAGCACCAAAATATTGTTTTCTTCACCAACCATCAGTTCATAGGTCCATTCTCTTAGCGCAACTACAGAGAACAGGGGCACAAACAGGCAAAAAACGGGCACAACCTCAATGGAGTGATGCAACCTGCCTGGAGTAAATGATGACACAAGGCAATTGACCCACGCATGTATCTATCTCATTTTCTTACACCTTCTATTACCTTCTGCTCTCTCTGATTTGGAAAAAGCTGAAAAAAAAGGTTGAAACCAGTTCCCTGAAATTATTCCCCTACTTGACTAATAAGTATATAAAGACGGTAGGTATTGATTGTAATTCTGTAAATCTATTTCTTAAACTTCTTAAATTCTACTTTTATAGTTAGTCTTTTTTTTAGTTTTAAAACACCAAGAACTTAGTTTCGAATAAACACACATAAACAAACAAAGGATCCATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTAGGCCTTACTAGTAGCGGCCGCTGCAG

GAATTCGCGGCCGCTTCTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCAG

GGTCTCAATCTAAGTTTTAATTACAAAGGATCCATGAGATTCCCTAGTATCTTCACCGCCGTCTTGTTCGCTGCTAGTTCTGCTTTGGCTGCTCCAGTCAACACTACCACCGAAGACGAAACCGCTCAGATCCCAGCTGAAGCCGTCATCGGTTACCTTGACCTTGAGGGTGACTTCGATGTCGCCGTCCTTCCATTCTCTAACAGTACCAACAACGGTCTTTTGTTCATTAACACCACCATTGCTTCTATCGCCGCTAAGGAGGAGGGTGTCTCTCTTGACAAGAGACATCACCACCACCATCACAGTGCTAAGTCTAGTTTCATCTCTACCACCACTACCGACCTTACCTCTATCAACACCAGTGCCTACAGTACCGGTTCTATCTCTACCGTCGAAACCGGAAACAGAACCACCTCTGAGGTCATCTCTCATGTCGTCACCACCTCTACCAAGCTTAGTCCTACCGCCACCACCAGTCTTACCATCGCTCAAACTTCTATTTATAGTACCGACTCTAATATTACCGTCGGTACCGACATCCACACCACCAGTGAAGTCATCTCTGACGTCGAAACCATCTCTAGGGAAACCGCCTCTACCGTTGTTGCCGCCCCAACTTCTACCACTGGTTGGACCGGTGCCATGAACACTTACATCTCTCAATTCACTTCTTCTAGTTTTGCTACTATTAACTCTACCCCTATCATCTCTTCTAGTGCCGTCTTCGAAACCAGTGACGCCTCTATCGTCAACGTCCATACCGAGAACATCACCAACACTGCTGCCGTTCCATCTGAGGAGCCTACCTTCGTCAACGCCACTAGAAACTCTCTTAATTCTTTTTGTTCTTCTAAACAACCATCTTCTCCTAGTTCTTATACTTCTTCTCCTTTGGTTTCTTCTTTGTCTGTTTCTAAGACCTTGTTGTCTACCTCTTTCACCCCTTCTGTCCCAACTTCTAATACTTATATTAAGACCAAGAACACTGGTTACTTCGAGCACACTGCCTTGACCACCTCTAGTGTTGGATTGAATAGTTTCTCTGAAACCGCCGTCTCTAGTCAAGGTACCAAGATTGACACCTTTTTGGTTTCTAGTTTGATCGCCTACCCATCTTCTGCCTCTGGTTCTCAGCTTAGTGGTATCCAGCAAAATTTTACCTCTACTTCTTTGATGATCAGTACCTACGAGGGTAAGGCCAGTATCTTCTTCTCTGCCGAGTTGGGTTCTATTATTTTTTTGTTGCTTAGTTATTTGTTGTTTTAAAACGGGTACTGTACAGTTAGTACATTGAGTCGAAATATACGAAATTATTGTTCATAATTTTCATCCTGGCTCTTTTTTTCTTCAACCATAGTTAAATGGACAGTTCATATCTTAACTCTAATAATACTTTTCTAGTTCTTATCCTTTTCCGTCTCACCGCAGATTTTATCATAGTATTAAATTTATATTTTGTTCGTAAAAAGAAAAATTTGTGAGCGTTACCGCTCGTTTCATTACCCGAAGGCTGTTTCAGTAGACCACTGATTAAGTAAGTAGATGAAAAAATTTCATCACCATGAAAGAGTTCGATGAGAGCTACTTTTTCAAATGCTTAACAGCTAACCGCCATTCAATAATGTTACGCTCTCTTCATTCTGCGGCTACGTTATCTAACAAGAGGTTTTACTCTCTCATATCTCATTCAAATAGAAAGAACATAATCAAAAGGATCCTCATGTAATTAGTTATGTCACGAGACC

GGTCTCACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAAGGATCCATAAAGCAATCTTGATGAGGAGAGACC

GGTCTCAAACACACATAAACAAACAAAGGATCCATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAAGGATCCATTGAATTGAATTGAAATCGAGAGACC

GGTCTCACGAACTATAATTAACTAAACGGATCCATAACTTCGTATAATGTATGCTATACGAAGTTATATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAATAACTTCGTATAATGTATGCTATACGAAGTTATGGATCCAGTGCTTTTAACTAAGAATTAGAGACC

2019.6.1 ~ 6.10

Construction plasmids：

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAAGGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAGGCCTTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCCTTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCAGTTCGAGTTTATCATTATCAATACTGCCATTTCAAAGAATACGTAAATAATTAATAGTAGTGATTTTCCTAACTTTATTTAGTCAAAAAATTAGCCTTTTAATTCTGCTGTAACCCGTACATGCCCAAAATAGGGGGCGGGTTACACAGAATATATAACATCGTAGGTGTCTGGGTGAACAGTTTATTCCTGGCATCCACTAAATATAATGGAGCCCGCTTTTTAAGCTGGCATCCAGAAAAAAAAAGAATCCCAGCACCAAAATATTGTTTTCTTCACCAACCATCAGTTCATAGGTCCATTCTCTTAGCGCAACTACAGAGAACAGGGGCACAAACAGGCAAAAAACGGGCACAACCTCAATGGAGTGATGCAACCTGCCTGGAGTAAATGATGACACAAGGCAATTGACCCACGCATGTATCTATCTCATTTTCTTACACCTTCTATTACCTTCTGCTCTCTCTGATTTGGAAAAAGCTGAAAAAAAAGGTTGAAACCAGTTCCCTGAAATTATTCCCCTACTTGACTAATAAGTATATAAAGACGGTAGGTATTGATTGTAATTCTGTAAATCTATTTCTTAAACTTCTTAAATTCTACTTTTATAGTTAGTCTTTTTTTTAGTTTTAAAACACCAAGAACTTAGTTTCGAATAAACACACATAAACAAACAAAGGATCCATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTAGGCCTTACTAGTAGCGGCCGCTGCA

We insert promoter TEF1 and terminator CYC1, promoter GPM1 and terminator TDH1, promoter TDH3 and terminator PGK1, promoter TEF2 and terminator ENO2 into vector pSB1C3 using restriction endonuclease and T4 DNA ligase, respectively.

The pSB1C3 digestion system we used:

pSB1C3 15 μL
EcoRI 3 μL
PstI 3 μL
Cutsmart Buffer 5 μL
ddH2O 24 μL


The enzyme system we use:

pSB1C3 2 μL
fragment 6 μL
T4 DNA Ligase 0.5 μL
T4 Buffer 1 μL
ddH2O 0.5 μL

BBa_k3126001:

BBa_k3126002:

BBa_k3126003:

BBa_k3126004:

2019.6.10 ~ 6.20

Construction plasmids：

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAAGGATCCATGAGATTCCCTAGTATCTTCACCGCCGTCTTGTTCGCTGCTAGTTCTGCTTTGGCTGCTCCAGTCAACACTACCACCGAAGACGAAACCGCTCAGATCCCAGCTGAAGCCGTCATCGGTTACCTTGACCTTGAGGGTGACTTCGATGTCGCCGTCCTTCCATTCTCTAACAGTACCAACAACGGTCTTTTGTTCATTAACACCACCATTGCTTCTATCGCCGCTAAGGAGGAGGGTGTCTCTCTTGACAAGAGACATCACCACCACCATCACAGTGCTAAGTCTAGTTTCATCTCTACCACCACTACCGACCTTACCTCTATCAACACCAGTGCCTACAGTACCGGTTCTATCTCTACCGTCGAAACCGGAAACAGAACCACCTCTGAGGTCATCTCTCATGTCGTCACCACCTCTACCAAGCTTAGTCCTACCGCCACCACCAGTCTTACCATCGCTCAAACTTCTATTTATAGTACCGACTCTAATATTACCGTCGGTACCGACATCCACACCACCAGTGAAGTCATCTCTGACGTCGAAACCATCTCTAGGGAAACCGCCTCTACCGTTGTTGCCGCCCCAACTTCTACCACTGGTTGGACCGGTGCCATGAACACTTACATCTCTCAATTCACTTCTTCTAGTTTTGCTACTATTAACTCTACCCCTATCATCTCTTCTAGTGCCGTCTTCGAAACCAGTGACGCCTCTATCGTCAACGTCCATACCGAGAACATCACCAACACTGCTGCCGTTCCATCTGAGGAGCCTACCTTCGTCAACGCCACTAGAAACTCTCTTAATTCTTTTTGTTCTTCTAAACAACCATCTTCTCCTAGTTCTTATACTTCTTCTCCTTTGGTTTCTTCTTTGTCTGTTTCTAAGACCTTGTTGTCTACCTCTTTCACCCCTTCTGTCCCAACTTCTAATACTTATATTAAGACCAAGAACACTGGTTACTTCGAGCACACTGCCTTGACCACCTCTAGTGTTGGATTGAATAGTTTCTCTGAAACCGCCGTCTCTAGTCAAGGTACCAAGATTGACACCTTTTTGGTTTCTAGTTTGATCGCCTACCCATCTTCTGCCTCTGGTTCTCAGCTTAGTGGTATCCAGCAAAATTTTACCTCTACTTCTTTGATGATCAGTACCTACGAGGGTAAGGCCAGTATCTTCTTCTCTGCCGAGTTGGGTTCTATTATTTTTTTGTTGCTTAGTTATTTGTTGTTTTAAAACGGGTACTGTACAGTTAGTACATTGAGTCGAAATATACGAAATTATTGTTCATAATTTTCATCCTGGCTCTTTTTTTCTTCAACCATAGTTAAATGGACAGTTCATATCTTAACTCTAATAATACTTTTCTAGTTCTTATCCTTTTCCGTCTCACCGCAGATTTTATCATAGTATTAAATTTATATTTTGTTCGTAAAAAGAAAAATTTGTGAGCGTTACCGCTCGTTTCATTACCCGAAGGCTGTTTCAGTAGACCACTGATTAAGTAAGTAGATGAAAAAATTTCATCACCATGAAAGAGTTCGATGAGAGCTACTTTTTCAAATGCTTAACAGCTAACCGCCATTCAATAATGTTACGCTCTCTTCATTCTGCGGCTACGTTATCTAACAAGAGGTTTTACTCTCTCATATCTCATTCAAATAGAAAGAACATAATCAAAAGGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAGGCCTTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCCTTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCAGTTCGAGTTTATCATTATCAATACTGCCATTTCAAAGAATACGTAAATAATTAATAGTAGTGATTTTCCTAACTTTATTTAGTCAAAAAATTAGCCTTTTAATTCTGCTGTAACCCGTACATGCCCAAAATAGGGGGCGGGTTACACAGAATATATAACATCGTAGGTGTCTGGGTGAACAGTTTATTCCTGGCATCCACTAAATATAATGGAGCCCGCTTTTTAAGCTGGCATCCAGAAAAAAAAAGAATCCCAGCACCAAAATATTGTTTTCTTCACCAACCATCAGTTCATAGGTCCATTCTCTTAGCGCAACTACAGAGAACAGGGGCACAAACAGGCAAAAAACGGGCACAACCTCAATGGAGTGATGCAACCTGCCTGGAGTAAATGATGACACAAGGCAATTGACCCACGCATGTATCTATCTCATTTTCTTACACCTTCTATTACCTTCTGCTCTCTCTGATTTGGAAAAAGCTGAAAAAAAAGGTTGAAACCAGTTCCCTGAAATTATTCCCCTACTTGACTAATAAGTATATAAAGACGGTAGGTATTGATTGTAATTCTGTAAATCTATTTCTTAAACTTCTTAAATTCTACTTTTATAGTTAGTCTTTTTTTTAGTTTTAAAACACCAAGAACTTAGTTTCGAATAAACACACATAAACAAACAAAGGATCCATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAAGGATCCATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTAGGCCTTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

Insertion of gene into plasmid carrying promoter and terminator.

BBa_K3126005 (pSB1C3-pTEF1/ tCYC1/ hexa His):

Enzyme digestion system:

BBa_K312600120 μL
BamHI 3 μL
buffer5 μL
ddH2O22 μL

Seamless cloning system:

BBa_K3126001 2 μL
hexa-His4 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

BBa_K3126006 (pSB1C3-pGPM1/tTDH1/ nixA):

Enzyme digestion system:

BBa_K312600220 μL
BamHI 3 μL
buffer5 μL
ddH2O22 μL

Seamless cloning system:

BBa_K3126002 2 μL
nixA4 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

BBa_K3126007 (pSB1C3-pTDH3/tPGK1/ TgMTP1t2):

Enzyme digestion system:

BBa_K312600320 μL
BamHI 3 μL
buffer5 μL
ddH2O22 μL

Seamless cloning system:

BBa__K3126003 2 μL
TgMTP1t24 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

BBa_K3126008 (pSB1C3-pTEF2/tENO2/ kanR):

Enzyme digestion system:

BBa_K312600320 μL
BamHI 3 μL
buffer5 μL
ddH2O22 μL

Seamless cloning system:

BBa_K3126004 2 μL
kanR4 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

2019.6.20 ~ 6.30

Construction of plasmids for the translation and expression of genes in S. cerevisiae:

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAAGGATCCATGAGATTCCCTAGTATCTTCACCGCCGTCTTGTTCGCTGCTAGTTCTGCTTTGGCTGCTCCAGTCAACACTACCACCGAAGACGAAACCGCTCAGATCCCAGCTGAAGCCGTCATCGGTTACCTTGACCTTGAGGGTGACTTCGATGTCGCCGTCCTTCCATTCTCTAACAGTACCAACAACGGTCTTTTGTTCATTAACACCACCATTGCTTCTATCGCCGCTAAGGAGGAGGGTGTCTCTCTTGACAAGAGACATCACCACCACCATCACAGTGCTAAGTCTAGTTTCATCTCTACCACCACTACCGACCTTACCTCTATCAACACCAGTGCCTACAGTACCGGTTCTATCTCTACCGTCGAAACCGGAAACAGAACCACCTCTGAGGTCATCTCTCATGTCGTCACCACCTCTACCAAGCTTAGTCCTACCGCCACCACCAGTCTTACCATCGCTCAAACTTCTATTTATAGTACCGACTCTAATATTACCGTCGGTACCGACATCCACACCACCAGTGAAGTCATCTCTGACGTCGAAACCATCTCTAGGGAAACCGCCTCTACCGTTGTTGCCGCCCCAACTTCTACCACTGGTTGGACCGGTGCCATGAACACTTACATCTCTCAATTCACTTCTTCTAGTTTTGCTACTATTAACTCTACCCCTATCATCTCTTCTAGTGCCGTCTTCGAAACCAGTGACGCCTCTATCGTCAACGTCCATACCGAGAACATCACCAACACTGCTGCCGTTCCATCTGAGGAGCCTACCTTCGTCAACGCCACTAGAAACTCTCTTAATTCTTTTTGTTCTTCTAAACAACCATCTTCTCCTAGTTCTTATACTTCTTCTCCTTTGGTTTCTTCTTTGTCTGTTTCTAAGACCTTGTTGTCTACCTCTTTCACCCCTTCTGTCCCAACTTCTAATACTTATATTAAGACCAAGAACACTGGTTACTTCGAGCACACTGCCTTGACCACCTCTAGTGTTGGATTGAATAGTTTCTCTGAAACCGCCGTCTCTAGTCAAGGTACCAAGATTGACACCTTTTTGGTTTCTAGTTTGATCGCCTACCCATCTTCTGCCTCTGGTTCTCAGCTTAGTGGTATCCAGCAAAATTTTACCTCTACTTCTTTGATGATCAGTACCTACGAGGGTAAGGCCAGTATCTTCTTCTCTGCCGAGTTGGGTTCTATTATTTTTTTGTTGCTTAGTTATTTGTTGTTTTAAAACGGGTACTGTACAGTTAGTACATTGAGTCGAAATATACGAAATTATTGTTCATAATTTTCATCCTGGCTCTTTTTTTCTTCAACCATAGTTAAATGGACAGTTCATATCTTAACTCTAATAATACTTTTCTAGTTCTTATCCTTTTCCGTCTCACCGCAGATTTTATCATAGTATTAAATTTATATTTTGTTCGTAAAAAGAAAAATTTGTGAGCGTTACCGCTCGTTTCATTACCCGAAGGCTGTTTCAGTAGACCACTGATTAAGTAAGTAGATGAAAAAATTTCATCACCATGAAAGAGTTCGATGAGAGCTACTTTTTCAAATGCTTAACAGCTAACCGCCATTCAATAATGTTACGCTCTCTTCATTCTGCGGCTACGTTATCTAACAAGAGGTTTTACTCTCTCATATCTCATTCAAATAGAAAGAACATAATCAAAAGGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAGGCCTTACTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCCTTACTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCAGTTCGAGTTTATCATTATCAATACTGCCATTTCAAAGAATACGTAAATAATTAATAGTAGTGATTTTCCTAACTTTATTTAGTCAAAAAATTAGCCTTTTAATTCTGCTGTAACCCGTACATGCCCAAAATAGGGGGCGGGTTACACAGAATATATAACATCGTAGGTGTCTGGGTGAACAGTTTATTCCTGGCATCCACTAAATATAATGGAGCCCGCTTTTTAAGCTGGCATCCAGAAAAAAAAAGAATCCCAGCACCAAAATATTGTTTTCTTCACCAACCATCAGTTCATAGGTCCATTCTCTTAGCGCAACTACAGAGAACAGGGGCACAAACAGGCAAAAAACGGGCACAACCTCAATGGAGTGATGCAACCTGCCTGGAGTAAATGATGACACAAGGCAATTGACCCACGCATGTATCTATCTCATTTTCTTACACCTTCTATTACCTTCTGCTCTCTCTGATTTGGAAAAAGCTGAAAAAAAAGGTTGAAACCAGTTCCCTGAAATTATTCCCCTACTTGACTAATAAGTATATAAAGACGGTAGGTATTGATTGTAATTCTGTAAATCTATTTCTTAAACTTCTTAAATTCTACTTTTATAGTTAGTCTTTTTTTTAGTTTTAAAACACCAAGAACTTAGTTTCGAATAAACACACATAAACAAACAAAGGATCCATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAAGGATCCATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTAGGCCTTACTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

The resistance gene(kanR) was linked to the target gene(hexa-His, nixA, TgMTP1t2) to screen the engineering yeast and the original yeast.

BBa_K3126009 (pSB1C3-pTEF1/ tCYC1/ hexa-His/ kanR):

Enzyme digestion system:

BBa_K312600525 μL
SpeI 3 μL
PstI 3 μL
Cutsmart Buffer5 μL
ddH2O14 μL

BBa_K312600825 μL
XbaI 3 μL
PstI 3 μL
Cutsmart Buffer5 μL
ddH2O14 μL

The enzyme system we use:

BBa_K3126005 linear plasmid 2 μL
kanR part6 μL
T4 DNA Ligase0.5 μL
T4Buffer1 μL
ddH2O0.5 μL

BBa_K3126010 (pSB1C3-pGPM1/tTDH1/ nixA/ kanR)

Enzyme digestion system:

BBa_K312600625 μL
SpeI 3 μL
PstI 3 μL
Cutsmart Buffer5 μL
ddH2O14 μL

BBa_K312600825 μL
XbaI 3 μL
PstI 3 μL
Cutsmart Buffer5 μL
ddH2O14 μL

The enzyme system we use:

BBa_K3126006 linear plasmid 2 μL
kanR part4 μL
T4 DNA Ligase0.5 μL
T4Buffer1 μL
ddH2O0.5 μL

BBa_K3126011 (pSB1C3-pTDH3/tPGK1/ TgMTP1t2/ kanR)

Enzyme digestion system:

BBa_K312600725 μL
SpeI 3 μL
PstI 3 μL
Cutsmart Buffer5 μL
ddH2O14 μL

BBa_K312600825 μL
XbaI 3 μL
PstI 3 μL
Cutsmart Buffer5 μL
ddH2O14 μL

The enzyme system we use:

BBa_K3126007 linear plasmid 2 μL
kanR part4 μL
T4 DNA Ligase0.5 μL
T4Buffer1 μL
ddH2O0.5 μL

2019.7.1 ~ 7.10

Construction of plasmids for the translation and expression of genes in S. cerevisiae:

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAAGGATCCATGAGATTCCCTAGTATCTTCACCGCCGTCTTGTTCGCTGCTAGTTCTGCTTTGGCTGCTCCAGTCAACACTACCACCGAAGACGAAACCGCTCAGATCCCAGCTGAAGCCGTCATCGGTTACCTTGACCTTGAGGGTGACTTCGATGTCGCCGTCCTTCCATTCTCTAACAGTACCAACAACGGTCTTTTGTTCATTAACACCACCATTGCTTCTATCGCCGCTAAGGAGGAGGGTGTCTCTCTTGACAAGAGACATCACCACCACCATCACAGTGCTAAGTCTAGTTTCATCTCTACCACCACTACCGACCTTACCTCTATCAACACCAGTGCCTACAGTACCGGTTCTATCTCTACCGTCGAAACCGGAAACAGAACCACCTCTGAGGTCATCTCTCATGTCGTCACCACCTCTACCAAGCTTAGTCCTACCGCCACCACCAGTCTTACCATCGCTCAAACTTCTATTTATAGTACCGACTCTAATATTACCGTCGGTACCGACATCCACACCACCAGTGAAGTCATCTCTGACGTCGAAACCATCTCTAGGGAAACCGCCTCTACCGTTGTTGCCGCCCCAACTTCTACCACTGGTTGGACCGGTGCCATGAACACTTACATCTCTCAATTCACTTCTTCTAGTTTTGCTACTATTAACTCTACCCCTATCATCTCTTCTAGTGCCGTCTTCGAAACCAGTGACGCCTCTATCGTCAACGTCCATACCGAGAACATCACCAACACTGCTGCCGTTCCATCTGAGGAGCCTACCTTCGTCAACGCCACTAGAAACTCTCTTAATTCTTTTTGTTCTTCTAAACAACCATCTTCTCCTAGTTCTTATACTTCTTCTCCTTTGGTTTCTTCTTTGTCTGTTTCTAAGACCTTGTTGTCTACCTCTTTCACCCCTTCTGTCCCAACTTCTAATACTTATATTAAGACCAAGAACACTGGTTACTTCGAGCACACTGCCTTGACCACCTCTAGTGTTGGATTGAATAGTTTCTCTGAAACCGCCGTCTCTAGTCAAGGTACCAAGATTGACACCTTTTTGGTTTCTAGTTTGATCGCCTACCCATCTTCTGCCTCTGGTTCTCAGCTTAGTGGTATCCAGCAAAATTTTACCTCTACTTCTTTGATGATCAGTACCTACGAGGGTAAGGCCAGTATCTTCTTCTCTGCCGAGTTGGGTTCTATTATTTTTTTGTTGCTTAGTTATTTGTTGTTTTAAAACGGGTACTGTACAGTTAGTACATTGAGTCGAAATATACGAAATTATTGTTCATAATTTTCATCCTGGCTCTTTTTTTCTTCAACCATAGTTAAATGGACAGTTCATATCTTAACTCTAATAATACTTTTCTAGTTCTTATCCTTTTCCGTCTCACCGCAGATTTTATCATAGTATTAAATTTATATTTTGTTCGTAAAAAGAAAAATTTGTGAGCGTTACCGCTCGTTTCATTACCCGAAGGCTGTTTCAGTAGACCACTGATTAAGTAAGTAGATGAAAAAATTTCATCACCATGAAAGAGTTCGATGAGAGCTACTTTTTCAAATGCTTAACAGCTAACCGCCATTCAATAATGTTACGCTCTCTTCATTCTGCGGCTACGTTATCTAACAAGAGGTTTTACTCTCTCATATCTCATTCAAATAGAAAGAACATAATCAAAAGGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAGGGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCCTTACTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCAGTTCGAGTTTATCATTATCAATACTGCCATTTCAAAGAATACGTAAATAATTAATAGTAGTGATTTTCCTAACTTTATTTAGTCAAAAAATTAGCCTTTTAATTCTGCTGTAACCCGTACATGCCCAAAATAGGGGGCGGGTTACACAGAATATATAACATCGTAGGTGTCTGGGTGAACAGTTTATTCCTGGCATCCACTAAATATAATGGAGCCCGCTTTTTAAGCTGGCATCCAGAAAAAAAAAGAATCCCAGCACCAAAATATTGTTTTCTTCACCAACCATCAGTTCATAGGTCCATTCTCTTAGCGCAACTACAGAGAACAGGGGCACAAACAGGCAAAAAACGGGCACAACCTCAATGGAGTGATGCAACCTGCCTGGAGTAAATGATGACACAAGGCAATTGACCCACGCATGTATCTATCTCATTTTCTTACACCTTCTATTACCTTCTGCTCTCTCTGATTTGGAAAAAGCTGAAAAAAAAGGTTGAAACCAGTTCCCTGAAATTATTCCCCTACTTGACTAATAAGTATATAAAGACGGTAGGTATTGATTGTAATTCTGTAAATCTATTTCTTAAACTTCTTAAATTCTACTTTTATAGTTAGTCTTTTTTTTAGTTTTAAAACACCAAGAACTTAGTTTCGAATAAACACACATAAACAAACAAAGGATCCATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAAGGATCCATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTAGGCCTTACTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATAACTTCGTATAATGTATGCTATACGAAGTTATATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAATAACTTCGTATAATGTATGCTATACGAAGTTATGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

GTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGCTTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCATTACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGTCTCATGTCTATCCCAGAAACTCAAAAAGGTGTTATCTTCTACGAATCCCCCTTGCCAACAGGGAGTTCTTCAGAGACATGGAGGCTCAAAACGAAATTATTGACAGCCTAGACATCAATAGTCATACAACAGAAAGCGACCACCCAACTTTGGCTGATAATAGCGTATAAACAATGCATACTTTGTACGTTCAAAATACAATGCAGTAGATATATTTATGCATATTACATATAATACATATCACATAGGAAGCAACAGGCGCGTTGGACTTTTAATTTTCGAGGACCGCGAATCCTTACATCACACCCAATCCCCCACAAGTGATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGACACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCATCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAAGGATCCATGAGATTCCCTAGTATCTTCACCGCCGTCTTGTTCGCTGCTAGTTCTGCTTTGGCTGCTCCAGTCAACACTACCACCGAAGACGAAACCGCTCAGATCCCAGCTGAAGCCGTCATCGGTTACCTTGACCTTGAGGGTGACTTCGATGTCGCCGTCCTTCCATTCTCTAACAGTACCAACAACGGTCTTTTGTTCATTAACACCACCATTGCTTCTATCGCCGCTAAGGAGGAGGGTGTCTCTCTTGACAAGAGACATCACCACCACCATCACAGTGCTAAGTCTAGTTTCATCTCTACCACCACTACCGACCTTACCTCTATCAACACCAGTGCCTACAGTACCGGTTCTATCTCTACCGTCGAAACCGGAAACAGAACCACCTCTGAGGTCATCTCTCATGTCGTCACCACCTCTACCAAGCTTAGTCCTACCGCCACCACCAGTCTTACCATCGCTCAAACTTCTATTTATAGTACCGACTCTAATATTACCGTCGGTACCGACATCCACACCACCAGTGAAGTCATCTCTGACGTCGAAACCATCTCTAGGGAAACCGCCTCTACCGTTGTTGCCGCCCCAACTTCTACCACTGGTTGGACCGGTGCCATGAACACTTACATCTCTCAATTCACTTCTTCTAGTTTTGCTACTATTAACTCTACCCCTATCATCTCTTCTAGTGCCGTCTTCGAAACCAGTGACGCCTCTATCGTCAACGTCCATACCGAGAACATCACCAACACTGCTGCCGTTCCATCTGAGGAGCCTACCTTCGTCAACGCCACTAGAAACTCTCTTAATTCTTTTTGTTCTTCTAAACAACCATCTTCTCCTAGTTCTTATACTTCTTCTCCTTTGGTTTCTTCTTTGTCTGTTTCTAAGACCTTGTTGTCTACCTCTTTCACCCCTTCTGTCCCAACTTCTAATACTTATATTAAGACCAAGAACACTGGTTACTTCGAGCACACTGCCTTGACCACCTCTAGTGTTGGATTGAATAGTTTCTCTGAAACCGCCGTCTCTAGTCAAGGTACCAAGATTGACACCTTTTTGGTTTCTAGTTTGATCGCCTACCCATCTTCTGCCTCTGGTTCTCAGCTTAGTGGTATCCAGCAAAATTTTACCTCTACTTCTTTGATGATCAGTACCTACGAGGGTAAGGCCAGTATCTTCTTCTCTGCCGAGTTGGGTTCTATTATTTTTTTGTTGCTTAGTTATTTGTTGTTTTAAAACGGGTACTGTACAGTTAGTACATTGAGTCGAAATATACGAAATTATTGTTCATAATTTTCATCCTGGCTCTTTTTTTCTTCAACCATAGTTAAATGGACAGTTCATATCTTAACTCTAATAATACTTTTCTAGTTCTTATCCTTTTCCGTCTCACCGCAGATTTTATCATAGTATTAAATTTATATTTTGTTCGTAAAAAGAAAAATTTGTGAGCGTTACCGCTCGTTTCATTACCCGAAGGCTGTTTCAGTAGACCACTGATTAAGTAAGTAGATGAAAAAATTTCATCACCATGAAAGAGTTCGATGAGAGCTACTTTTTCAAATGCTTAACAGCTAACCGCCATTCAATAATGTTACGCTCTCTTCATTCTGCGGCTACGTTATCTAACAAGAGGTTTTACTCTCTCATATCTCATTCAAATAGAAAGAACATAATCAAAAGGATCCTCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAGGGCCAAACTTTTCGGTTAACACATGCAGTGATGCACGCGCGATGGTGCTAAGTTACATATATATATATATATATATATATATATATATATAGCCATAGTGATGTCTAAGTAACCTTTATGGTATATTTCTTAATGTGGAAAGATACTAGCGCGCGCACCCACACACAAGCTTCGTCTTTTCTTGAAGAAAAGAGGAAGCTCGCTAAATGGGATTCCACTTTCCGTTCCCTGCCAGCTGATGGAAAAAGGTTAGTGGAACGATGAAGAATAAAAAGAGAGATCCACTGAGGTGAAATTTCAGCTGACAGCGAGTTTCATGATCGTGATGAACAATGGTAACGAGTTGTGGCTGTTGCCAGGGAGGGTGGTTCTCAACTTTTAATGTATGGCCAAATCGCTACTTGGGTTTGTTATATAACAAAGAAGAAATAATGAACTGATTCTCTTCCTCCTTCTTGTCCTTTCTTAATTCTGTTGTAATTACCTTCCTTTGTAATTTTTTTTGTAATTATTCTTCTTAATAATCCAAACAAACACACATATTACAATAGGATCCATGATTAAGTACCATGTCATGAAAGAAGGTTACATCTTGACTGTTTTTAAAAATGAAAGGTTGAGTTGGTTGCCATATATCGCCATCGTCATTTTGTTGCACGTTATTGGTTTCTCTTTCTTGTGGATCGCCGGTAAGGACCATCACATTTTGTTCGGTATGGGTATCCTTGCCTACACTCTTGGTTTGAGGCATGCCTTTGATGCCGACCATATCGCCGCTATCGACAACACCGTTAGAAAGTTGTTGCAGCAGAGAAAGGACCCATCTGGTGTCGGTTTCTACTTCTCTATTGGACATTCTTCTGTCGTCTTCTTGATGGCCGTTTTCCTTGGTGTCTCTGTCAAGTGGGCCAAGGACGAACTTCCTCACTTCCAAGATATCGGAGGTACCATTGGAACCCTTGTTTCTGGTTTTTTCTTGGTTCTTATTGGAGTTCTTAATCTTATTATTCTTATTTCTTTGATTAACTTGTTCGCCAAGTTGAGGAGGGAACACATCGAAGAAGCTGAGGTCGATGCCTTGTTGGAGTCTAGGGGTCTTGTCTCTAGGTTCGTCGGACCATACTTTAAGTTGATCACCAGAAGTTGGCACGTCTTGCCTCTTGGATTCCTTTTCGGATTGGGATTCGACACCGCTTCTGAGATCGCCTTGCTTGCCCTTAGTAGTGGAGCCTCTCAGCAAGCTATCTCTTTCATTGGTATTTTGTCTTTGCCTATCTTGTTCGCCAGTGGTATGAGTCTTCTTGACACCCTTGACGGTGTCGTCATGAAGTACGCCTACAACTGGGCCTTCTTCAACCCTATCAGAAAAATCTATTACAACATTACCATTACCGCCATCTCTGTTATGGCCGCCCTTGTTATCGGTATGATCGAATTGCTTCAAATTCTTGCCGACAAACTTGACTTGCATGGAGCCTTCTGGGCCTTCATTGGAAGTATCGAGTTCGATTATTTGGGTTATATCTTGGTTGCCTTGTTTCTTATTACTTGGTTGATCAGTTCTTTGATTTGGAAATTTGGAAGAATTGAACACAAATGGAGTAGATAAGGATCCATAAAGCAATCTTGATGAGGATAATGATTTTTTTTTGAATATACATAAATACTACCGTTTTTCTGCTAGATTTTGTGATGACGTAAATAAGTACATATTACTTTTTAAGCCAAGACAAGATTAAGCATTAACTTTACCCTTTTCTTTCTAAGTTTCAATATTAGTTATCACTGTTTAAAAGTTATGGCGAGAACGTCGGCGGTTAAAATATATTACCCTGAACGAGGCAGTTCGAGTTTATCATTATCAATACTGCCATTTCAAAGAATACGTAAATAATTAATAGTAGTGATTTTCCTAACTTTATTTAGTCAAAAAATTAGCCTTTTAATTCTGCTGTAACCCGTACATGCCCAAAATAGGGGGCGGGTTACACAGAATATATAACATCGTAGGTGTCTGGGTGAACAGTTTATTCCTGGCATCCACTAAATATAATGGAGCCCGCTTTTTAAGCTGGCATCCAGAAAAAAAAAGAATCCCAGCACCAAAATATTGTTTTCTTCACCAACCATCAGTTCATAGGTCCATTCTCTTAGCGCAACTACAGAGAACAGGGGCACAAACAGGCAAAAAACGGGCACAACCTCAATGGAGTGATGCAACCTGCCTGGAGTAAATGATGACACAAGGCAATTGACCCACGCATGTATCTATCTCATTTTCTTACACCTTCTATTACCTTCTGCTCTCTCTGATTTGGAAAAAGCTGAAAAAAAAGGTTGAAACCAGTTCCCTGAAATTATTCCCCTACTTGACTAATAAGTATATAAAGACGGTAGGTATTGATTGTAATTCTGTAAATCTATTTCTTAAACTTCTTAAATTCTACTTTTATAGTTAGTCTTTTTTTTAGTTTTAAAACACCAAGAACTTAGTTTCGAATAAACACACATAAACAAACAAAGGATCCATGGAGAGTTCTTCTCATATTATTGAAGTCAACGGAGGTAGATCTGACGAGGAAAGAAGGGCTGTCGCTAGTAAGGTCTGCGGAGAAGCTCCTTGCGGTTTCAGTGACGCTAAGAACGCTTCTGGTGACGCCCAAGAAAGGAACGCCTCTATGAGGAAATTGTGCATCGCCGTCGTCTTGTGCCTTGTCTTCATGTCTGTCGAGGTCGTTGGTGGTGTCAAGGCCAACAGTTTGGCCATCATGACCGATGCCGCTCATCTTTTGAGTGACGTTGCCGCTTTCGCCATCTCTTTGTTCAGTCTTTGGGCTGCCGGTTGGGAGGCTACCCCAAGGCACACCTACGGATTCTTCAGAATCGAAATTCTTGGAGCTCTTGTTTCTATCCAATTGATTTGGTTGCTTACTGGTATCCTTGTTTACGAGGCCATCATTAGACTTTTGACCGAAACCTCTGAGGTCAACGGTTTCTTGATGTTCGCTGTCGCCACTTTCGGTTTGCTTGTCAACATTATCATGGCTGTCATGCTTGGTCACGACCATGACAATGGTCACGGTCATAGTCACGGTGAGGACAAACACGATGAAGCCCATGGTGACGTCACTGAACAGCTTCTTGAAAAGCCAAAACAGCAGAAGGAAAAAAAGAAGAGGAACATCAACGCCCAAGGTGCCTATTTGCACGTCTTGGGTGACTCTATCCAGTCTGTCGGAGTCATGATCGGTGGTGCTGCTATCTGGTACAATCCAAAGTGGAAGATCATTGACTTGATCTGCACCTTGGTCTTCTCTGTCATCGTCTTGGGAACCACCATCAATATGATCAGAAGTATTCTTGAGGTCTTGATGGAGAGTACCCCAAGAGAAATCGACGCCACCAAGCTTGAGAAGGGTCTTCTTGAGATGGAGGAGGTCGTCGCTGTTCACGAATTGCACATCTGGGCCTCTACCGTTGGTAAGGTCTTGCTTGCTTGCCATGTCAACGCCAGACCAGATGCCGATGCCGATATGGTCCTTAACAAAGTCGTTGACTATATTAGGAGAGAATATAACATTTCTCATGTTACTATCCAAATTGAAAGATAAGGATCCATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTAGGCCTTACTAGAGTTGATAGGTCAAGATCAATGTAAACAATTACTTTGTTATGTAGAGTTTTTTTAGCTACCTATATTCCACCATAACATCAATCATGCGGTTGCTGGTGTATTTACCAATAATGTTTAATGTATATATATATATATATATATGGGGCCGTATACTTACATATAGTAGATGTCAAGCGTAGGCGCTTCCCCTGCCGGCTGTGAGGGCGCCATAACCAAGGTATCTATAGACCGCCAATCAGCAAACTACCTCCGTACATTCATGTTGCACCCACACATTTATACACCCAGACCGCGACAAATTACCCATAAGGTTGTTTGTGACGGCGTCGTACAAGAGAACGTGGGAACTTTTTAGGCTCACCAAAAAAGAAAGAAAAAATACGAGTTGCTGACAGAAGCCTCAAGAAAAAAAAAATTCTTCTTCGACTATGCTGGAGGCAGAGATGATCGAGCCGGTAGTTAACTATATATAGCTAAATTGGTTCCATCACCTTCTTTTCTGGTGTCGCTCCTTCTAGTGCTATTTCTGGCTTTTCCTATTTTTTTTTTTCCATTTTTCTTTCTCTCTTTCTAATATATAAATTCTCTTGCATTTTCTATTTTTCTCTCTATCTATTCTACTTGTTTATTCCCTTCAAGGTTTTTTTTTAAGGAGTACTTGTTTTTAGAATATACGGTCAACGAACTATAATTAACTAAACGGATCCATAACTTCGTATAATGTATGCTATACGAAGTTATATGGGAAAGGAGAAGACCCACGTCAGTAGACCAAGGCTTAACAGTAACATGGACGCCGACTTGTACGGATACAAGTGGGCCAGAGACAACGTCGGTCAGTCTGGAGCTACCATCTATAGACTTTACGGAAAGCCAGACGCCCCAGAGTTGTTCCTTAAGCACGGTAAGGGATCTGTTGCCAACGACGTCACCGATGAGATGGTTAGACTTAACTGGCTTACCGAGTTCATGCCACTTCCAACCATCAAGCACTTCATTAGAACCCCAGATGACGCTTGGTTGTTGACCACTGCCATCCCCGGTAAGACCGCCTTTCAAGTTTTGGAAGAGTACCCAGACTCTGGAGAGAACATCGTTGACGCCTTGGCCGTCTTCCTTAGGAGGTTGCACTCTATCCCAGTTTGCAACTGCCCATTCAATAGTGATAGAGTCTTCAGACTTGCCCAAGCCCAGTCTAGGATGAACAACGGATTGGTCGATGCCTCTGACTTCGACGACGAGAGAAATGGATGGCCAGTCGAGCAAGTTTGGAAGGAGATGCATAAGCTTTTGCCTTTTTCTCCAGACAGTGTTGTCACCCACGGTGACTTTAGTTTGGACAACCTTATTTTCGACGAGGGTAAATTGATCGGTTGCATCGACGTCGGTAGAGTTGGTATCGCCGATAGATACCAAGACTTGGCCATCTTGTGGAACTGCTTGGGTGAGTTCTCTCCAAGTCTTCAAAAAAGATTGTTCCAAAAATATGGTATTGACAACCCAGATATGAATAAATTGCAATTTCATTTGATGTTGGATGAGTTTTTCTAAATAACTTCGTATAATGTATGCTATACGAAGTTATGGATCCAGTGCTTTTAACTAAGAATTATTAGTCTTTTCTGCTTATTTTTTCATCATAGTTTAGAACACTTTATATTAACGAATAGTTTATGAATCTATTTAGGTTTAAAAATTGATACAGTTTTATAAGTTACTTTTTCAAAGACTCGTGCTGTCTATTGCATAATGCACTGGAAGGGGAAAAAAAAGGTGCACACGCGTGGCTTTTTCTTGAATTTGCAGTTTGAAAAATAAAGGGTCAAATCGTTGGTAGATACGTTGTTGACACTTCTAAATAAGAGACCTACTAGTAGCGGCCGCTGCA

We obtained the fragments to be inserted by PCR, then combine several target genes by seamless cloning.

BBa_K3126012 (pSB1C3- hexa-His/ nixA/ kanR):

The PCR system of pGPM1-nixA-tTDH1:

(P-nixA-T_F

CGAAGGCTTTAATTTGCAGGGCCAAACTTTTCGGTTAACA

P-nixA-T_R

ACCTATCAACTCTAGTAAGGCCTCGTTCAGGGTAATATATTTT)

BBa_K3126006/SpeⅠ PstⅠ1 μL
P-nixA-T_F 2 μL
P-nixA-T_R2 μL
2×PCR mix20 μL
ddH2O15 μL

Enzyme digestion system:

BBa_K312600915 μL
StuI 3 μL
buffer 5 μL
ddH2O27 μL

Seamless cloning system:

BBa_K3126009/StuⅠ 2 μL
pGPM1-nixA-tTDH14 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

BBa_K3126013 (pSB1C3- nixA/ TgMTP1t2/ kanR)

The PCR system of pTDH3-TgMTP1t2-tPGK1

BBa_K3126007/SpeⅠ PstⅠ1 μL
P-TgMTP1t2-T_F2 μL
P-TgMTP1t2-T_R2 μL
2×PCR mix20 μL
ddH2O15 μL

Enzyme digestion system:

BBa_K312601015 μL
Stu>I 3 μL
buffer 5 μL
ddH2O27 μL

Seamless cloning system:

BBa_K3126010/StuⅠ 2 μL
pTDH3-TgMTP1t2-tPGK14 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

BBa_K3126014 (pSB1C3- hexa-His/ nixA/ TgMTP1t2/ kanR)

Enzyme digestion system:

BBa_K312601215 μL
StuI 3 μL
buffer5 μL
ddH2O27 μL

Seamless cloning system:

BBa_K3126012/StuⅠ2 μL
pTDH3-TgMTP1t2-tPGK1 4 μL
5×CEⅡ Buffer2 μL
Exnase Ⅱ1 μL
ddH2O1 μL

2019.7.20 ~ 7.25

The plasmid is digested with Bsa1 to get the fragment to be integrated.

pSB1C3- hexa-His- kanR/ BsaⅠ

Enzyme digestion system:

BBa_K312600925μL
BsaI 3 μL
buffer5 μL
ddH2O17 μL

pSB1C3-nixA- kanR/ BsaⅠ

Enzyme digestion system:

BBa_K3126001025μL
BsaI 3 μL
buffer5 μL
ddH2O17 μL

pSB1C3-TgMTP1t2- kanR/ BsaⅠ

Enzyme digestion system:

BBa_K3126001125μL
BsaI 3 μL
buffer5 μL
ddH2O17 μL

pSB1C3-hexa-His-nixA- kanR/ BsaⅠ

Enzyme digestion system:

BBa_K3126001225μL
BsaI 3 μL
buffer5 μL
ddH2O17 μL

pSB1C3-nixA-TgMTP1t2- kanR/ BsaⅠ

Enzyme digestion system:

BBa_K3126001325μL
BsaI 3 μL
buffer5 μL
ddH2O17 μL

pSB1C3-hexa-His-nixA-TgMTP1t2- kanR/ BsaⅠ

Enzyme digestion system:

BBa_K3126001425μL
BsaI 3 μL
buffer5 μL
ddH2O17 μL

2019.7.25 ~ 8.5

The fragments were transformed into S. cerevisiae to express the corresponding protein.

option date

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