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[求助]
求助：请问构建这个质粒时选择了EcoRI和BamHI位点，设计引物时怎样看是否会发生移码？已有1人参与
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载体为：PBV220,酶切位点为EcoRI和BamHI（上游、下游），请问插入目的基因片段时如何看是否发生移码？万分感谢！ 质粒序列为： 1 gaattcccgg ggatccgtcg acctgcagcc aagcttggct gttttggcgg atgagagaag 61 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg 121 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc 181 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca 241 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt 301 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg 361 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa 421 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa 481 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 541 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 601 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 661 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 721 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 781 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt 841 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 901 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 961 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 1021 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 1081 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 1141 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 1201 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 1261 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 1321 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 1381 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 1441 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 1501 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 1561 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 1621 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 1681 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 1741 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 1801 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 1861 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 1921 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 1981 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 2041 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 2101 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 2161 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 2221 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 2281 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 2341 gcgaggaagc ggaagagcgc ttatctttcc ctttattttt gctgcggtaa gtcgcataaa 2401 aaccattctt cataattcaa tccatttact atgttatgtt ctgaggggag tgaaaattcc 2461 cctaattcga tgaagattct tgctcaattg ttatcagcta tgcgccgacc agaacacctt 2521 gccgatcagc caaacgtctc ttcaggccac tgactagcga taactttccc cacaacggaa 2581 caactctcat tgcatgggat cattgggtac tgtgggttta gtggttgtaa aaacacctga 2641 ccgctatccc tgatcagttt cttgaaggta aactcatcac ccccaagtct ggctatgcag 2701 aaatcacctg gctcaacagc ctgctcaggg tcaacgagaa ttaacattcc gtcaggaaag 2761 cttggcttgg agcctgttgg tgcggtcatg gaattacctt caacctcaag ccagaatgca 2821 gaatcactgg cttttttggt tgtgcttacc catctctccg catcaccttt ggtaaaggtt 2881 ctaagcttag gtgagaacat ccctgcctga acatgagaaa aaacagggta ctcatactca 2941 cttctaagtg acggctgcat actaaccgct tcatacatct cgtagatttc tctggcgatt 3001 gaagggctaa attcttcaac gctaactttg agaatttttg taagcaatgc ggcgttataa 3061 gcatttaatg cattgatgcc attaaataaa gcaccaacgc ctgactgccc catccccatc 3121 ttgtctgcga cagattcctg ggataagcca agttcatttt tctttttttc ataaattgct 3181 ttaaggcgac gtgcgtcctc aagctgctct tgtgttaatg gtttcttttt tgtgctcata 3241 cgttaaatct atcaccgcaa gggataaata tctaacaccg tgcgtgttga ctattttacc 3301 tctggcggtg ataatggttg catgtactaa ggaggttgta tggaacaacg cataaccctg 3361 aaagattatg caatgcgctt tgggcaaacc aagacagcta aagatctctc acctaccaaa 3421 caatgccccc ctgcaaaaaa taaattcata taaaaaacat acagataacc atctgcggtg 3481 ataaattatc tctggcggtg ttgacataaa taccactggc ggtgatactg agcacatcag 3541 caggacgcac tgaccaccat gaaggtgacg ctcttaaaaa ttaagccctg aagaagggca 3601 gcattcaaag cagaaggctt tggggtgtgt gatacgaaac gaagcattgg ttaaaaatta 3661 aggagg  pBV220.png |
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