pRS314

pRS314

pRS314载体基本信息：

pRS314载体图谱：

pRS314载体简介：

YCtype centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ) peptide. pRSS56, constructed by ligating a PvuI fragment (bp 4982412) of pBluescript KS+ to a PvuI fragment (bp 2850 730) of pBS(+), contains the KS MCS from pBluescript KS(+) and the unique NdeI and AatII sites between bla and f1 origin of pBS(+). A genomic HindII/PstI fragment (1.002 kb) containing the TRP1 gene was inserted into the NdeI site and a cassette containing CEN6 and the ARS associated with histone 4 (ARSH4) was inserted into the AatII site of pRSS56. All ends were blunted. An EcoRI site in the TRP1containing fragment (external to the coding sequence) was destroyed. The order of the major features in this plasmid is: TRP1f1 ori (NaeI) T7 promoter lacZ/MCS T3 promoter pMB1 ori bla CEN6 ARSH4.

pRS314载体序列：

ORIGIN

        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt

       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata

      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt

      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa

      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata

      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct

      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata

      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact

      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa

      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg

      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg

      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt

      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt

      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg

      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa

      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt

      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag

     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt

     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga

     1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt

     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta

     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg

     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc

     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt

     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg

     1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg

     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa

     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa

     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga

     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg

     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact

     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac

     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg

     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg

     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga

     2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc

     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg

     2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc

     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc

     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt

     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc

     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc

     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac

     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact

     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg

     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt

     2761 aaccctcact aaagggaaca aaagctgggt accgggcccc ccctcgaggt cgacggtatc

     2821 gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgcc

     2881 accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt

     2941 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca

     3001 tccccccttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca

     3061 gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc

     3121 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc

     3181 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa

     3241 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact

     3301 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt

     3361 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa

     3421 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt

     3481 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac

     3541 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaggcaa

     3601 gtgcacaaac aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg

     3661 acgaaatttg ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca

     3721 tcaacaccaa taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca

     3781 ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc

     3841 gagttccaat ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac

     3901 gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt

     3961 cttttaataa ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc

     4021 agttggacga tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga

     4081 ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca

     4141 agacttgaaa ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat

     4201 ataataccca gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc

     4261 aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc

     4321 caagctgcct ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct

     4381 ccctcttggc cctctccttt tcttttttcg accgaattaa ttcttaatcg gcaaaaaaag

     4441 aaaagctccg gatcaagatt gtacgtaagg tgacaagcta tttttcaata aagaatatct

     4501 tccactactg ccatctggcg tcataactgc aaagtacaca tatattacga tgctgtctat

     4561 taaatgcttc ctatattata tatatagtaa tgtcgtttat ggtgcactct cagtacaatc

     4621 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc

     4681 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc

     4741 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga

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