pLysS质粒

基本信息

启动子:	T7 Phi-3.8
复制子:	p15 ori
质粒分类:	有毒蛋白表达配套质粒
质粒大小:	4886bp
质粒标签:	T7 lysozyme
原核抗性:	氯霉素Chloramphenicol
克隆菌株:	DH5α
培养条件:	37℃，有氧，LB
表达宿主:	大肠杆菌
培养条件:	参考相关文献
5'测序引物:	根据序列设计
3'测序引物:	根据序列设计
备注:	低拷贝

质粒简介

         pLysS and pLysE are 4886bp plasmids constructed by insertion of the T7 lysozyme gene into the BamH I site of pACYC184 (1, 2). These plasmids are not cloning vectors; they are used in λDE3 lysogenic hosts to suppress basal expression from the T7 promoter by producing T7 lysozyme, a natural inhibitor of T7 RNA polymerase. The two plasmids differ only by the orientation of the T7 lysozyme gene. In pLysS the T7 lysozyme coding sequence is in the antisense orientation relative to the tet promoter, so only a small amount of T7 lysozyme is produced. In pLysE large amounts of T7 lysozyme are produced from the tet promoter. The construct also contains the weak T7 φ3.8 promoter immediately following the lysozyme gene. The p15A origin of replication is compatible with those found in pBR322- and pUC-derived plasmids.Unique sites are shown on the circle map.
        1. Studier , F.W. (1991) J. Mol. Biol. 219, 37-44.
        2. Chang, A.C.Y. and Cohen, S.N. (1978) J. Bacteriol. 134, 1141.

质粒图谱

质粒序列

LOCUS       Exported                4886 bp ds-DNA     circular SYN 10-SEP-2016
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pLysS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4886)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..4886
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(220..322)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1392)
                     /direction=LEFT
                     /note="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1504..1532
                     /note="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        complement(1981..2014)
                     /note="T7 Phi-3.8 promoter"
                     /note="weak promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2012..2467)
                     /codon_start=1
                     /product="lysozyme from bacteriophage T7"
                     /note="T7 lysozyme"
                     /note="inhibitor of T7 RNA polymerase"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAGLRAHHEVAPKACPSFDLKRWWEKNELVTSDRG"
     CDS             complement(4446..219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct
      961 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1021 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1081 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1141 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1201 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1261 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1321 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1381 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1441 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     1501 tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta
     1561 acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt
     1621 caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga
     1681 tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt
     1741 gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc
     1801 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
     1861 cgtcctgtgg atccggccca ttggctgcct cccacacttg gatatgcctc ctcggagcct
     1921 tatagaattg tttataagac ttgcgcatta tttgacctcc aatgcgaaca aagggaaacc
     1981 gctgtggtct ccctttagtg agttcaatta attatccacg gtcagaagtg accagttcgt
     2041 tcttctccca ccaacgctta aggtcgaacg aagggcaagc cttcggcgcc acctcatgat
     2101 gggcgcgaag accagcgcct tcgtacttag ccagcagtgt gacaagcagt gagcgaaggg
     2161 attgcatttg ggctggcgta aagttagcgt cgaacttacc tttatcgtcg ataccaccaa
     2221 caaggcagac gccgatagag ttgtggttgt aacccttagc gtgagagcct acagccatct
     2281 catctcgtcc tgcctccaca gtaccgtctc gcttgatgat aaagtggtat cccacatcga
     2341 gccaaccctg ctctttgtgc cactggcgaa tctcacggac accaacattc tgacttggct
     2401 tggtagccga gcagtgaaca aagattgcgt cagtagattc acgttgttta aactgtacac
     2461 gagccattat ttctttcctc ctttcctttt taatctatca aaggggaccc ggatcctcta
     2521 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     2581 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     2641 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     2701 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     2761 aatgcaggag tcgcataagg gagagcgtcg accgatgccc ttgagagcct tcaacccagt
     2821 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     2881 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     2941 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     3001 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     3061 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     3121 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     3181 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     3241 cgcggctctt accagcctaa cttcgatcac tggaccgctg atcgtcacgg cgatttatgc
     3301 cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt
     3361 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     3421 cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg
     3481 agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc
     3541 agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     3601 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     3661 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     3721 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtcc
     3781 cctacgtgct gctgaagttg cccgcaacag agagtggaac caaccggtga taccacgata
     3841 ctatgactga gagtcaacgc catgagcggc ctcatttctt attctgagtt acaacagtcc
     3901 gcaccgctgt ccggtagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     3961 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgcc caacagtccc
     4021 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgccctgcac cattatgttc
     4081 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     4141 aagcgctaac cgtttttatc aggctctggg aggcagaata aatgatcata tcgtcaatta
     4201 ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag cacacggtca
     4261 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     4321 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     4381 cttattatca cttattcagg cgtagcacca ggcgtttaag ggcaccaata actgccttaa
     4441 aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     4501 ccgacatgga agccatcaca gacggcatga tgaacctgaa tcgccagcgg catcagcacc
     4561 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     4621 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     4681 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     4741 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     4801 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     4861 agctcaccgt ctttcattgc catacg
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