求救！引物设计
丁香园论坛
2053
我设计的引物如下：
上游引物加BamHI CGGGA TCCTGTCGCA CCAAATCGTA 73.3 62.6
下游引物加 XbaI
GCTCTA GACATATGCC ACCAGCAACT AACATG 70.4 65.6
Two primers complementarity.
Max complementarity in continuous: 4 bp, free energy= 0.20 Kcal/mol
5'-GCTCTAGACATATGCCACCAGCAACTAACATG-3'
||||
3'-ATGCTAAACCACGCTGTCCTAGGGC-5'
Max complementarity in discontinuous: 9 bp
5'-GCTCTAGACATATGCCACCAGCAACTAACATG-3'
|| | | ||| | |
3'-ATGCTAAACCACGCTGTCCTAGGGC-5'
请指点： 1引物行吗？
2扩pcr用多少的退火温度？
3加接头的pcr程序不一样吗？要分几步吗？
以下是我要扩的片段！可以在前500和后700之间设计引物！那位高手帮忙一下，我要加Xba I 和BamH I 的接头！哪位高手指导一下！谢谢！
序列：
1 GACATGTGGA ATTTTACGCC ACCAAAAACT GCCCGCAAAA TAACTAGTTT CGCTGCTGCT ACGGAAACAG CCAAATCACA GATTCCTCTC GGGGGATGAT
101 GGCAAAAGAA TAAAATCATT TAAGCGAAGT ATTGCCCACA AACCTCTGAA GAGACAAGAG AACCGAGGCG GTTCCAGGCT GCAAAGATAA CAGCAGCCAG
201 GGGTTGTGAA CATGCTGGGG GCCAAACATC CCTCCCCCCC CCCTGCTCCA TCACATTTTT AATTCCCTTG CTTCGTTTCC ATAGGGGGAA GGCCAAGTTG
301 ATTATCACCG ACCCTTTTTG GAGCCAACAA CACTTTTTCC CCAGAATTTT TGTTTTCTCT TCTTCTTCTC GCATCCCCTT TTTGTTCCCA GTCCTGTCGC
401 ACCAAATCGT ACGGAACACT TACCACATTC GAGTATCCGT TTTGTGCAAC ACACCACCGC GCGGCCGTTC TGTTGAGCAA CGTCGGCCCG AGAGGCGAAA
501 AAAATGAAAA CCCAAATCTA GGTAGTCAGG GTGAGGAGCA GAACGTTTGA CATCCAACCG TCTTGCTGTT AGTGAACAAG TGCCAAGATC GTAGTACCGA
601 ATCTAGGGTA GTAGGAACAA GTAAAAAAAG GAAGCCATCA TCACAAGCCC TTGCAGTAAA GCCACATCAG CCCACCCCGT CCAAAATGAG AAAAAAGTCC
701 CATTGAGCTG TGCCGCCGCG TCAGGCCAAA CACCAAGAAT TGGGGGGAAG CCTTTAAACT AATGCAATCG CGAAGGCAGT TTTCTTCTTC TCTTTTTCTG
801 AGTTCCCGTT CAGCTTTGCT CCACAAAGTG GCTCTATCCC TGTTTGTCAA CGCCAGAGGC CCGGAGCGTG GTCCGGGAAT TTTTTCTCTC GCGTTCCCCA
901 CTTTGCGCGG CAGCCATTGG AGAAGGGGGG AGGGAAGAGA AGGGGAAAAA GAAAATCTAT GACAAAACAG TGACCCACTG GCGGGGGAAG AGGAGAAGAA
1001 AAAAGGTACC CCTCCCCAGC CTCCCTGTTT TCTTATTTTC TGGTGTTTGC TGGTCGCAAA AATTTGGGTC TCGTCGTTTG TGGTGGTGCT TTTTTTTGTT
1101 TTTATCTTTT TTTCCCCCCT GGTTGAGGTG AGAGCCAGGT GAGTTTGGGG TCGGCCCCAT GGAGAGCGGG CAAAAATCCA GGGACTGACG AGTAGGCGTC
1201 GATGGATGCG TGTCGTGGGT GGGGTCACCC CTACACCACC CCCCCGGTGG CAGCCCATTT TGGCTGGTGC AAACGGTGGG GGCCCTGCTT GACGGTGGAT
1301 GATGGTCCAT CAGACGTCAA CCTTATTTGG ACATCTGCCA AGCTCAGCTT TCACTTACAC CGAGCATGGG AGAGACGCAA GGGATGGGAA TGAAGACAAC
1401 AGAATTCCAG AAGCCGCGGA AAGACGGAAG CGAAGCATAT AATAAATGAC GGTCTGTCCT GTTCAAAAAA ACATCTATCA ATTTCCCGGT TTCCCGCCTG
1501 CTCCATTGTT AGTGTTTCTT ACCAGGACCT GTAACCCAAT TTCTTGGGGA CCGCAAGGTA AAGAAATCAA ACTCCCCATT TTAGATTATG TCGAAGAGAA
1601 AATTTATGGA CGTTGTTTTT CTCGCAAGAT TTTTCAGTTA TCTCTAACGA GGAGTCGCCC GCACATAGGT ACGTTTGCAT GTTAGCGTGA CCGTGCGCCT
1701 GTCTCTCGCG CAAACAAAGA GCGAGAAGCC AGACCCACTC AAAACTGCTG CCATTGGTGC TTGCCACTTT CTATTCAGCC TTCGATCCTC TATTGAGATT
1801 TCTATTGAGC TTCTCACTTT CTATTCAGAA TCTCATTTTC TATTCAGGTT TATTAAGTTA TTGAGGAACC AACTGGGGAC CCCACTTCGT TTCTTTATTA
1901 ACTCGACCTT CGAACCCAGG AACCCAACTC CCTCCAAGAC TCCGACATCA CCAAAACCAC AAGACTCTTT CCATTCGATT GCGCTAGAAC TACTCACAAA
2001 CCTCAGATTG CAAACACAAA CCCCATCGTC AGATCATTGA AACGAGGCAC ATCTACAATG GATTCATTGC AGATGGACTC CTCTCAAATG GAGGCGTTGT
2101 CGTCGATTTG GGTCATCGTG CAGCATCAGC TCGCCCAAAT CAACAATTCC AACAACATAG CTCATATCGG CGCCGTTCTC CTCAACCAAC TTGAGGCGCC
2201 TTTCAGGAGC ACTTTGACAT CATGGTTTCA GCAGTAAGTT CTCCATAAGC TCAAGGCTTT TCCGCAAGGG AAACTAACAT CAGGTAGGCA CACCGGCATC
2301 AACCAGGTCT CAGTCGTGCG CGACATCAAG GCTCGTCGAC ACTACATCGG ACCAACTGAT GCTTTTCAGG AGAGCTATCA AGTTGTACAC ATCCAGGGTC
2401 ACGGATTTGT CTGGCTTTGG GATCTCCAGG TTGATGGTGA GGATGCCAGC ATCAATTCCT GCAATATCTC TGCCGAGACA GCACAGAACT TTTCCCAGCT
2501 GCGTCTTGAC GAGATGGCGC CTACCAATGA TGTTACGATG GTAGGTGATG AGAGCCTCAT CAACGGCAAC GGCATTGACT TTGCCATGTT GGAAGACATC
2601 GGAAGCCCCG CATCAAACGG CTCCTCATTC CCGAGCCCTC AAGTTTTCAC CCGGGAAATT TCATTCGATG ATCTACCAGT TGGCATTATG ACTGCCGAGG
2701 CCATTCAGAG AGACCTTCTT GGCGCCAATA TTCGCCCGGC TAACCTCATT GATCCGAGTC TTGCGAGCAA TCAGAATGTC CACAACATCA TCCATCAGAA
2801 TAACATTGCC CAGGCTTTTA CAGATCACCT GAACATGATG GCATCCAATG CTGTCCACGA TGACACAGAA GAGGGTCTGA CAGCCGCGGA TCTTATGCCG
2901 GCGTTGCATC GCATGTTCGT GCCGGGTCTT GAGTCAATTG CAATGGAGCA GTTCCACACC ACGCTTCCTA ATGGCGCACT CCTTCACCTG TCAATTCCTG
3001 TCGATGAACC TGAGGATGCT GTCTCGACCA CCGCGCCTGC TGCTCAACAG ACTTCCTTGC AAGACAAGAT CCCTCGTCCT GCGAATGCCT ACATCCTGTA
3101 CCGCAAGGAT TGGCATCCCA TAGTCAAGTC TGCAAATCCA GGAATCCATA ACAACGAAAT CTGTGAGTTC TTTTACCTCC AAACTTCCCT AGCTGCTGCT
3201 AACTTTTTTT CCTTTAGCAA AAATCCTGGG GAAGCAGTGG GCAGCTGAGA CACCGGAAGT GCGGGCTGAG TACAAGGAGC TCGCGGAGGA GAAGAAGAGA
3301 GAGTTCTACG CAAAGTACCC TACATATCGC TACTCTCCTC GTCGGCCGAG TGAGATAATG CGCCGAGTGA GTTTGTAACA CATTATTGCG TGAAAGAGTG
3401 GACAGCGAGC TAACAAATTA CTTGCCAACA AGAACACGAA AAGCCGAAAG ACCCCAACGA ACGGGGCAGC GAAGCCGAGC AAAGCAAAGA AGTGCGGCAA
3501 CGGGGTTACT GGTGGAGCAG ACAGTGCAGT CGATGTCAGC TCCCCGGGAT CGCCATTTTA GATGAGGTCA CCATTCACAT CATCCGAGAT ACATTGTACA
3601 AGTCAGGAGC AGTTATCAAA GCTGCATTTC ACCAGAGTCC TCATATCTGA TTTTTTCTTC TTTCTTTTCA CATTGCCCCA TCGTACATCC AGAGAGCTGG
3701 ACTCGAAATA CCCTCTCAAT AGAGTCTGCA AAGAAGATCT TAGGACCTAC GACATAGGTT ACGTGGTCAT TATTCACCAA GAACTACATA TCTGCCAGCG
3801 AAACATATTC AGTTTTCAGA GCCATTTGTC GAAAGAAATT TTGAGATGCC GGCCCTCTGG TCACCAGTCA AAGTTTGCGG CATCGTTTCG GAGCTCAGGT
3901 AAGCCGACCC AGAGTCCTCA CACCAGAGCG ACAGCTCTTC AGCATCTAAC GATCACAGGC TAACACTTTT GGCGCCTCAA TTAGCAAGAA CATGGCCTGC
4001 ATGGTTTGGA TCTGGGTCGT CTGCGTTCCT CAGAAGCGGT ACCGCATCAC GACAGCTCGC TGGAACTCCA GTGGGCTGCT GGTTGGCCGC GTTGATTGTG
4101 AGTTTTCTAC TTTCTTCACC TTTTCCTTCC GTCCCAGATC TGAGCAGAGA CTGACAAGCG ACACAAAACC TCCCCACAGC TCCTCCTCAA TCTGCCACCA
4201 CAGCGTCCTC ACAGTCATCG TCATTCGACG GCAGTGAGGA AGACACAAAC ACAAACATCA ACGCCAATGG TATCGATGAG AGCCTGCTGC CGCCTGCTCT
4301 CCGCGGCCGC ACTGGACCTT CCAAGCCTCC AAACCGCTGG ATTCTCTACC GTGCCGCCAA GTCAGCTGAG CTGCGGGCAG ACAACCCATC CATGAACGCG
4401 AGCGAAATCT GTAAGTTCTT CGCTCTTTTC ACTCTTTTTC TCCGCTACCA CTTGTACCTA CCTTTTCCCC ATCTCCCATA CTCTTTGAAA AGTCTCCAGT
4501 CCCACCTCTC AAGCTGCTGT TTTGACAAGA TAAAAGACTA ACCAACATCT CTCTCCTCAC ACAGCTCAAG TTGCATCCCT GATGTGGCAG GCAGAGTCCG
4601 CCGCGACAAA GGCAGAGTGG GAGGAGCGCG CAGCCGAAGC CCGCGAGGAG CACATGCGAA ACACTCCAGA GTATGCGGTG GCGGCCGGTC TGCGCCAGCG
4701 CGCCATCGCC GAGAGGAGAA GGCAGAGGCA GCAGAAGAAG AAGCAACAGG AGCAGCAGAA GGGCGACGGT GACAAGGTGG CCAAGGAGAC TTCTACATCC
4801 ACAGAGAGCT TGGTCACTAC GGACGTCGGC GCTGTCAGCA ACCACGGCAG CAGCAGCAGT GAGGTGCCTG TCACAAGCCC ACACTCGCCG ACACTACTCA
4901 GTCCCGCGTC TGACAGCGAT TGTACGCCCG TCGGCAGCCC TCGAGCTCGC CACGGCTGGG GTTCTGATGC TCTCAGCCTG ATGCTCGACG CCGCCGAATT
5001 CTCTACTCTC TTCTGATCTG GTCTCTGGCA CTCTGAGCGT GCTATTCCCA GGAAATCAAA CACACAAAAA AAAAACCACA AAAAAAACTC CCCCCATTCA
5101 CATCCATCAT ACCCCTTCGG GACTTTCCGA CATGCATATA TCTCCCACCT CTGTTTTGTT TTTTTCCTTT TTCTCCTTTG TACTTGTCAT CTTTTTTCAT
5201 CTCAGGGTTT TGACCAGGAA CTGCCTACCT GGCTGCATAG GGACACGGCG CGATTATGAG TTTTGAGGAA ATGCTGGGGC AGCATCAGTC AGTATGCCCC
5301 TATTTTGTTC TCAAAGTTTA CTTGTTCTGT TTTTAGCTGG TTCTTCTGCA TCATTCACGG GAGAAGGGGA ATAAAGGAGT CTTCACAGGC ATATGACCCA
5401 TCATGACATA GATACCCTTA CGTTTTTCTT TTCTTTTCTT TTCTCAAGTC TCTCTGTTCT TTTACAGTTG CGCATCAGAT GGGATGACAC AAGATGTTAC
5501 CACGAGAAAT GCACTCAGCA AGTCGATACA CTAAGTCAAT CATTCGACCT GTTACCAACT CTAAAAACTC CTCGCTACGT GCCATTGTTT GTCCTAGTGA
5601 AACTGCCGAT GAGATATGTG TGATATGAGA TTACAATGGT GACAATGGTA ACAAGATGTG TCGTCCGCCC TGTAGCATAG TTTGCTTTGA CGTAGAAACC
5701 CAACGCCAGT AGCGGCACTC CGCCGCCTTA GGTCTGCTGT GACGCGTTCC AGTCGCTGCT CCAGATGAAT GTGCCGCATC TCCACTCGGT ACCTTTTTCC
5801 TTCTCGCCTG ACGGCCCAAG AGGTCTGCCG CAAATGTAAA AGGACCGGCC TGCATGACAA GAAGGTAAAC ATGTTAGTTG CTGGTGGCAT ATGTCACGAA
5901 GAGCCGTTTT AATACGTTGC TATCAAGATG CTGTCTCAAG CGGGCCACGA GGCGGCAAGC ATACAAGCTT GCATAAAGTA CAGACGATGA ATGAGGCAGA
6001 TAAAGACTAG GAAAATACGA GAAGAAACAT TTTCTGTCAG ACATACCACA ATTGATGCCT GGCTTCTTGG TCTGTAGGCT GATGCACGGT TCGTCGTGCT
上游引物加BamHI CGGGA TCCTGTCGCA CCAAATCGTA 73.3 62.6
下游引物加 XbaI
GCTCTA GACATATGCC ACCAGCAACT AACATG 70.4 65.6
Two primers complementarity.
Max complementarity in continuous: 4 bp, free energy= 0.20 Kcal/mol
5'-GCTCTAGACATATGCCACCAGCAACTAACATG-3'
||||
3'-ATGCTAAACCACGCTGTCCTAGGGC-5'
Max complementarity in discontinuous: 9 bp
5'-GCTCTAGACATATGCCACCAGCAACTAACATG-3'
|| | | ||| | |
3'-ATGCTAAACCACGCTGTCCTAGGGC-5'
请指点： 1引物行吗？
2扩pcr用多少的退火温度？
3加接头的pcr程序不一样吗？要分几步吗？
以下是我要扩的片段！可以在前500和后700之间设计引物！那位高手帮忙一下，我要加Xba I 和BamH I 的接头！哪位高手指导一下！谢谢！
序列：
1 GACATGTGGA ATTTTACGCC ACCAAAAACT GCCCGCAAAA TAACTAGTTT CGCTGCTGCT ACGGAAACAG CCAAATCACA GATTCCTCTC GGGGGATGAT
101 GGCAAAAGAA TAAAATCATT TAAGCGAAGT ATTGCCCACA AACCTCTGAA GAGACAAGAG AACCGAGGCG GTTCCAGGCT GCAAAGATAA CAGCAGCCAG
201 GGGTTGTGAA CATGCTGGGG GCCAAACATC CCTCCCCCCC CCCTGCTCCA TCACATTTTT AATTCCCTTG CTTCGTTTCC ATAGGGGGAA GGCCAAGTTG
301 ATTATCACCG ACCCTTTTTG GAGCCAACAA CACTTTTTCC CCAGAATTTT TGTTTTCTCT TCTTCTTCTC GCATCCCCTT TTTGTTCCCA GTCCTGTCGC
401 ACCAAATCGT ACGGAACACT TACCACATTC GAGTATCCGT TTTGTGCAAC ACACCACCGC GCGGCCGTTC TGTTGAGCAA CGTCGGCCCG AGAGGCGAAA
501 AAAATGAAAA CCCAAATCTA GGTAGTCAGG GTGAGGAGCA GAACGTTTGA CATCCAACCG TCTTGCTGTT AGTGAACAAG TGCCAAGATC GTAGTACCGA
601 ATCTAGGGTA GTAGGAACAA GTAAAAAAAG GAAGCCATCA TCACAAGCCC TTGCAGTAAA GCCACATCAG CCCACCCCGT CCAAAATGAG AAAAAAGTCC
701 CATTGAGCTG TGCCGCCGCG TCAGGCCAAA CACCAAGAAT TGGGGGGAAG CCTTTAAACT AATGCAATCG CGAAGGCAGT TTTCTTCTTC TCTTTTTCTG
801 AGTTCCCGTT CAGCTTTGCT CCACAAAGTG GCTCTATCCC TGTTTGTCAA CGCCAGAGGC CCGGAGCGTG GTCCGGGAAT TTTTTCTCTC GCGTTCCCCA
901 CTTTGCGCGG CAGCCATTGG AGAAGGGGGG AGGGAAGAGA AGGGGAAAAA GAAAATCTAT GACAAAACAG TGACCCACTG GCGGGGGAAG AGGAGAAGAA
1001 AAAAGGTACC CCTCCCCAGC CTCCCTGTTT TCTTATTTTC TGGTGTTTGC TGGTCGCAAA AATTTGGGTC TCGTCGTTTG TGGTGGTGCT TTTTTTTGTT
1101 TTTATCTTTT TTTCCCCCCT GGTTGAGGTG AGAGCCAGGT GAGTTTGGGG TCGGCCCCAT GGAGAGCGGG CAAAAATCCA GGGACTGACG AGTAGGCGTC
1201 GATGGATGCG TGTCGTGGGT GGGGTCACCC CTACACCACC CCCCCGGTGG CAGCCCATTT TGGCTGGTGC AAACGGTGGG GGCCCTGCTT GACGGTGGAT
1301 GATGGTCCAT CAGACGTCAA CCTTATTTGG ACATCTGCCA AGCTCAGCTT TCACTTACAC CGAGCATGGG AGAGACGCAA GGGATGGGAA TGAAGACAAC
1401 AGAATTCCAG AAGCCGCGGA AAGACGGAAG CGAAGCATAT AATAAATGAC GGTCTGTCCT GTTCAAAAAA ACATCTATCA ATTTCCCGGT TTCCCGCCTG
1501 CTCCATTGTT AGTGTTTCTT ACCAGGACCT GTAACCCAAT TTCTTGGGGA CCGCAAGGTA AAGAAATCAA ACTCCCCATT TTAGATTATG TCGAAGAGAA
1601 AATTTATGGA CGTTGTTTTT CTCGCAAGAT TTTTCAGTTA TCTCTAACGA GGAGTCGCCC GCACATAGGT ACGTTTGCAT GTTAGCGTGA CCGTGCGCCT
1701 GTCTCTCGCG CAAACAAAGA GCGAGAAGCC AGACCCACTC AAAACTGCTG CCATTGGTGC TTGCCACTTT CTATTCAGCC TTCGATCCTC TATTGAGATT
1801 TCTATTGAGC TTCTCACTTT CTATTCAGAA TCTCATTTTC TATTCAGGTT TATTAAGTTA TTGAGGAACC AACTGGGGAC CCCACTTCGT TTCTTTATTA
1901 ACTCGACCTT CGAACCCAGG AACCCAACTC CCTCCAAGAC TCCGACATCA CCAAAACCAC AAGACTCTTT CCATTCGATT GCGCTAGAAC TACTCACAAA
2001 CCTCAGATTG CAAACACAAA CCCCATCGTC AGATCATTGA AACGAGGCAC ATCTACAATG GATTCATTGC AGATGGACTC CTCTCAAATG GAGGCGTTGT
2101 CGTCGATTTG GGTCATCGTG CAGCATCAGC TCGCCCAAAT CAACAATTCC AACAACATAG CTCATATCGG CGCCGTTCTC CTCAACCAAC TTGAGGCGCC
2201 TTTCAGGAGC ACTTTGACAT CATGGTTTCA GCAGTAAGTT CTCCATAAGC TCAAGGCTTT TCCGCAAGGG AAACTAACAT CAGGTAGGCA CACCGGCATC
2301 AACCAGGTCT CAGTCGTGCG CGACATCAAG GCTCGTCGAC ACTACATCGG ACCAACTGAT GCTTTTCAGG AGAGCTATCA AGTTGTACAC ATCCAGGGTC
2401 ACGGATTTGT CTGGCTTTGG GATCTCCAGG TTGATGGTGA GGATGCCAGC ATCAATTCCT GCAATATCTC TGCCGAGACA GCACAGAACT TTTCCCAGCT
2501 GCGTCTTGAC GAGATGGCGC CTACCAATGA TGTTACGATG GTAGGTGATG AGAGCCTCAT CAACGGCAAC GGCATTGACT TTGCCATGTT GGAAGACATC
2601 GGAAGCCCCG CATCAAACGG CTCCTCATTC CCGAGCCCTC AAGTTTTCAC CCGGGAAATT TCATTCGATG ATCTACCAGT TGGCATTATG ACTGCCGAGG
2701 CCATTCAGAG AGACCTTCTT GGCGCCAATA TTCGCCCGGC TAACCTCATT GATCCGAGTC TTGCGAGCAA TCAGAATGTC CACAACATCA TCCATCAGAA
2801 TAACATTGCC CAGGCTTTTA CAGATCACCT GAACATGATG GCATCCAATG CTGTCCACGA TGACACAGAA GAGGGTCTGA CAGCCGCGGA TCTTATGCCG
2901 GCGTTGCATC GCATGTTCGT GCCGGGTCTT GAGTCAATTG CAATGGAGCA GTTCCACACC ACGCTTCCTA ATGGCGCACT CCTTCACCTG TCAATTCCTG
3001 TCGATGAACC TGAGGATGCT GTCTCGACCA CCGCGCCTGC TGCTCAACAG ACTTCCTTGC AAGACAAGAT CCCTCGTCCT GCGAATGCCT ACATCCTGTA
3101 CCGCAAGGAT TGGCATCCCA TAGTCAAGTC TGCAAATCCA GGAATCCATA ACAACGAAAT CTGTGAGTTC TTTTACCTCC AAACTTCCCT AGCTGCTGCT
3201 AACTTTTTTT CCTTTAGCAA AAATCCTGGG GAAGCAGTGG GCAGCTGAGA CACCGGAAGT GCGGGCTGAG TACAAGGAGC TCGCGGAGGA GAAGAAGAGA
3301 GAGTTCTACG CAAAGTACCC TACATATCGC TACTCTCCTC GTCGGCCGAG TGAGATAATG CGCCGAGTGA GTTTGTAACA CATTATTGCG TGAAAGAGTG
3401 GACAGCGAGC TAACAAATTA CTTGCCAACA AGAACACGAA AAGCCGAAAG ACCCCAACGA ACGGGGCAGC GAAGCCGAGC AAAGCAAAGA AGTGCGGCAA
3501 CGGGGTTACT GGTGGAGCAG ACAGTGCAGT CGATGTCAGC TCCCCGGGAT CGCCATTTTA GATGAGGTCA CCATTCACAT CATCCGAGAT ACATTGTACA
3601 AGTCAGGAGC AGTTATCAAA GCTGCATTTC ACCAGAGTCC TCATATCTGA TTTTTTCTTC TTTCTTTTCA CATTGCCCCA TCGTACATCC AGAGAGCTGG
3701 ACTCGAAATA CCCTCTCAAT AGAGTCTGCA AAGAAGATCT TAGGACCTAC GACATAGGTT ACGTGGTCAT TATTCACCAA GAACTACATA TCTGCCAGCG
3801 AAACATATTC AGTTTTCAGA GCCATTTGTC GAAAGAAATT TTGAGATGCC GGCCCTCTGG TCACCAGTCA AAGTTTGCGG CATCGTTTCG GAGCTCAGGT
3901 AAGCCGACCC AGAGTCCTCA CACCAGAGCG ACAGCTCTTC AGCATCTAAC GATCACAGGC TAACACTTTT GGCGCCTCAA TTAGCAAGAA CATGGCCTGC
4001 ATGGTTTGGA TCTGGGTCGT CTGCGTTCCT CAGAAGCGGT ACCGCATCAC GACAGCTCGC TGGAACTCCA GTGGGCTGCT GGTTGGCCGC GTTGATTGTG
4101 AGTTTTCTAC TTTCTTCACC TTTTCCTTCC GTCCCAGATC TGAGCAGAGA CTGACAAGCG ACACAAAACC TCCCCACAGC TCCTCCTCAA TCTGCCACCA
4201 CAGCGTCCTC ACAGTCATCG TCATTCGACG GCAGTGAGGA AGACACAAAC ACAAACATCA ACGCCAATGG TATCGATGAG AGCCTGCTGC CGCCTGCTCT
4301 CCGCGGCCGC ACTGGACCTT CCAAGCCTCC AAACCGCTGG ATTCTCTACC GTGCCGCCAA GTCAGCTGAG CTGCGGGCAG ACAACCCATC CATGAACGCG
4401 AGCGAAATCT GTAAGTTCTT CGCTCTTTTC ACTCTTTTTC TCCGCTACCA CTTGTACCTA CCTTTTCCCC ATCTCCCATA CTCTTTGAAA AGTCTCCAGT
4501 CCCACCTCTC AAGCTGCTGT TTTGACAAGA TAAAAGACTA ACCAACATCT CTCTCCTCAC ACAGCTCAAG TTGCATCCCT GATGTGGCAG GCAGAGTCCG
4601 CCGCGACAAA GGCAGAGTGG GAGGAGCGCG CAGCCGAAGC CCGCGAGGAG CACATGCGAA ACACTCCAGA GTATGCGGTG GCGGCCGGTC TGCGCCAGCG
4701 CGCCATCGCC GAGAGGAGAA GGCAGAGGCA GCAGAAGAAG AAGCAACAGG AGCAGCAGAA GGGCGACGGT GACAAGGTGG CCAAGGAGAC TTCTACATCC
4801 ACAGAGAGCT TGGTCACTAC GGACGTCGGC GCTGTCAGCA ACCACGGCAG CAGCAGCAGT GAGGTGCCTG TCACAAGCCC ACACTCGCCG ACACTACTCA
4901 GTCCCGCGTC TGACAGCGAT TGTACGCCCG TCGGCAGCCC TCGAGCTCGC CACGGCTGGG GTTCTGATGC TCTCAGCCTG ATGCTCGACG CCGCCGAATT
5001 CTCTACTCTC TTCTGATCTG GTCTCTGGCA CTCTGAGCGT GCTATTCCCA GGAAATCAAA CACACAAAAA AAAAACCACA AAAAAAACTC CCCCCATTCA
5101 CATCCATCAT ACCCCTTCGG GACTTTCCGA CATGCATATA TCTCCCACCT CTGTTTTGTT TTTTTCCTTT TTCTCCTTTG TACTTGTCAT CTTTTTTCAT
5201 CTCAGGGTTT TGACCAGGAA CTGCCTACCT GGCTGCATAG GGACACGGCG CGATTATGAG TTTTGAGGAA ATGCTGGGGC AGCATCAGTC AGTATGCCCC
5301 TATTTTGTTC TCAAAGTTTA CTTGTTCTGT TTTTAGCTGG TTCTTCTGCA TCATTCACGG GAGAAGGGGA ATAAAGGAGT CTTCACAGGC ATATGACCCA
5401 TCATGACATA GATACCCTTA CGTTTTTCTT TTCTTTTCTT TTCTCAAGTC TCTCTGTTCT TTTACAGTTG CGCATCAGAT GGGATGACAC AAGATGTTAC
5501 CACGAGAAAT GCACTCAGCA AGTCGATACA CTAAGTCAAT CATTCGACCT GTTACCAACT CTAAAAACTC CTCGCTACGT GCCATTGTTT GTCCTAGTGA
5601 AACTGCCGAT GAGATATGTG TGATATGAGA TTACAATGGT GACAATGGTA ACAAGATGTG TCGTCCGCCC TGTAGCATAG TTTGCTTTGA CGTAGAAACC
5701 CAACGCCAGT AGCGGCACTC CGCCGCCTTA GGTCTGCTGT GACGCGTTCC AGTCGCTGCT CCAGATGAAT GTGCCGCATC TCCACTCGGT ACCTTTTTCC
5801 TTCTCGCCTG ACGGCCCAAG AGGTCTGCCG CAAATGTAAA AGGACCGGCC TGCATGACAA GAAGGTAAAC ATGTTAGTTG CTGGTGGCAT ATGTCACGAA
5901 GAGCCGTTTT AATACGTTGC TATCAAGATG CTGTCTCAAG CGGGCCACGA GGCGGCAAGC ATACAAGCTT GCATAAAGTA CAGACGATGA ATGAGGCAGA
6001 TAAAGACTAG GAAAATACGA GAAGAAACAT TTTCTGTCAG ACATACCACA ATTGATGCCT GGCTTCTTGG TCTGTAGGCT GATGCACGGT TCGTCGTGCT
