[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

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

回复此楼