pJB866质粒

基本信息

启动子:	Pm promoter
复制子:	RK2 oriV
终止子:	Termnator
质粒分类:	其它宿主质粒；其它系列质粒；其它功能质粒
质粒大小:	8296bp
原核抗性:	四环素Tet
克隆菌株:	DH5α等大肠杆菌
培养条件:	30℃，有氧，LB
表达宿主:	革兰氏阴性菌
培养条件:	请参考文献
5'测序引物:	Pm-F：cggtttgatagggataagtcc
3'测序引物:	Ter-R：ccgcattaaaatctagcgagg

质粒简介

          pJB866是一个革兰氏阴性菌广宿主表达载体，Pm启动子驱动目的基因的表达。

质粒图谱

质粒序列

LOCUS       Exported                8296 bp ds-DNA     circular SYN 15-SEP-2017
DEFINITION  synthetic circular DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8296)
  TITLE     Direct Submission
  JOURNAL   Exported Sep 15, 2017 from Plasmid
FEATURES             Location/Qualifiers
     source          1..8296
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    5..127
                     /label=MCS
     promoter        complement(133..266)
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in 
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     CDS             966..1931
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator encoded by the Pseudomonas putida 
                     TOL plasmid pWWO"
                     /label=XylS
                     /note="stimulates transcription from the Pm promoter in the
                     presence of aromatic compounds such as m-toluate or 
                     benzoate"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGPPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     oriT            2471..2579
                     /note="incP origin of transfer"
     CDS             complement(3036..3686)
                     /codon_start=1
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHSTSVPRADDDWRSFLTGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSESGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             3717..4991
                     /codon_start=1
                     /label=TetA
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPDVKPNIPLIVILSTVALDAVG
                     IGLIMPVLPGLLRDLVHSNDVTAHYGILLALYALVQFACAPVLGALSDRFGRRPILLVS
                     LAGATVDYAIMATAPFLWVLYIGRIVAGITGATGAVAGAYIADITDGDERARHFGFMSA
                     CFGFGMVAGPVLGGLMGGFSPHAPFFAAAALNGLNFLTGCFLLPESHKGERRPLRREAL
                     NPLSFVRWARGMTVVAALMAVFFIMQLVGQVPAALWVIFGEDRFHWDATTIGISLAAFG
                     ILHSLAQAMITGPVAARLGERRALMLGMIADGTGYILLAFATRGWMAFPIMVLLASGGI
                     GMPALQAMLSRQVDEERQGQLQGSLAALTSLTSIVGPLLFTAIYAASITTWNGWAWIAG
                     AALYLLCLPALRRGLWSGAGQRADR"
     rep_origin      5566..6096
                     /label=RK2 oriV
                     /note="incP origin of replication"
     misc_feature    6318..6436
                     /label=Neo Promoter
     CDS             6488..7636
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    7687..8296
                     /label=Termnator
ORIGIN
        1 gatcttcgaa tgcatcgcgc gcaccgtacg tctcgaggaa ttcctgcagg atatctggat
       61 ccacgaagct tcccatggtg acgtcaccgg ttctagatac ctaggtgagc tctggtaccg
      121 cggccgcaat tcacatgttc atgactccat tattattgtt tctgttgcat aaagcctaag
      181 gggtaggcct ttctagagat agccattttt tgcactcctg tatccgcttc ttgcaaggct
      241 ggacttatcc ctatcaaacc ggacacccgg tcgttggtca gatcaagcta attcccatgc
      301 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
      361 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
      421 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
      481 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
      541 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgt
      601 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
      661 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagcagatc ccctttatcc
      721 gccaattcgt cgccgcatgc cccgacagca cgaacttctg gtgttctcgc ttcttaaaaa
      781 gaacgtcttc gttctgcttg gcgttatttt tgcttggaaa agtggtcact gattgcaaaa
      841 aggatggcgc aacgtggcaa tgggggtaac ccgtatacgc atcacgtcga gatgcatttt
      901 catcgacttg gcgcctttct acatcacacc aagcagccca cattaaaata agagaaccgt
      961 gaactatgga tttttgctta ttgaacgaga aaagtcagat cttcgtccac gccgagccct
     1021 atgcagtctc cgattatgtt aaccagtatg tcggtacgca ctctattcgc ctgcccaagg
     1081 gcgggccccc ggcaggcagg ctgcaccaca gaatcttcgg atgcctcgac ctgtgtcgaa
     1141 tcagctacgg cggtagcgtg agggtaatct cgcctggatt agagacctgt tatcatctgc
     1201 aaataatact caaaggccat tgcctgtggc gtggccatgg ccaggagcac tattttgcgc
     1261 cgggcgaact attgctgctc aatccggatg accaagccga cctgacctat tcagaagatt
     1321 gcgagaaatt tatcgttaaa ttgccctcag tggtccttga tcgggcatgc agtgacaaca
     1381 attggcacaa gccgagggag ggtatccgtt tcgccgcgcg acacaatctc cagcaactcg
     1441 atggctttat caatctactc gggttagttt gtgacgaagc ggaacataca aagtcgatgc
     1501 ctcgggtcca agagcactat gcggggatca tcgcttccaa gctgctcgaa atgctgggca
     1561 gcaatgtcag ccgtgaaatt ttcagcaaag gtaacccgtc tttcgagcga gtcgttcaat
     1621 tcattgagga gaatctcaaa cggaatatca gccttgagcg gttagcggag ctggcgatga
     1681 tgagtccacg ctcgctctac aatttgttcg agaagcatgc cggcaccacg ccgaagaact
     1741 acatccgcaa ccgcaagctc gaaagcatcc gcgcctgctt gaacgatccc agtgccaatg
     1801 tgcgtagtat aactgagata gccctagact acggcttctt acatttggga cgcttcgctg
     1861 aaaactatag gagcgcgttc ggcgagttgc cttccgacac cctgcgtcaa tgcaaaaagg
     1921 aagtggcttg attacgaacg tagccgaaga agggatgggt tggcatcgcc cggctttctt
     1981 agacactctc caagctctga aatagcgttt tacaaactcc actggctata gtgccggcgt
     2041 ttgcccgcgc tctccgtggc cgcgtcccaa tgcaggtcgg gttcctcgac ccgaagtgcc
     2101 cttgggcatc tccacttcag cgtctgacct tgctggccag ttcgccgccg atgccgaagt
     2161 ggcccttggc catctccgtg ataatcactc gcacgctggt cagcggcgca tccagggagc
     2221 gcgagatggc ctcgctgact tcccgaatga gggtttcctt ctgctcgtcg ctgcggcctt
     2281 caaggatgtg gatctgctgg ccgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2341 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatta attcttgaag
     2401 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2461 ttagagctta cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat
     2521 aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg
     2581 gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat
     2641 atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt
     2701 atgcagcgga aaagatccgt cggctgcagc gatccgtcga tctatctcat ctgcgcaagg
     2761 cagaacgtga agacggccgc cctggacctc gcccgcgagc gccagcgcac gaggccggcg
     2821 cgcggacccg ccgcggccca cgagcggacg ccgcagcagg agcgccagaa ggccgccaga
     2881 gaggccgagc gcggcgtgag gcttggacgc tagggcaggg catgaaaaag cccgtagcgg
     2941 gcgctacggg gctctgacgc ggtggaaagg gggaggggat gttgtctaca tggctctgct
     3001 gtagtgagtg ggttgcgctc cggcagcggt cctgatcaat cgtcaccctt tctcggtcct
     3061 tcaacgttcc tgacaacgag cctccttttc gccaatccat cgacaatcac cgcgagtccc
     3121 tgctcgaacg ctgcgtccgg accggcttcg tcgaaggcgt ctatcgcggc ccgcaacagc
     3181 ggcgagagcg gagcctgttc aacggtgccg ccgcgctcgc cggactcgct gtcgccggcc
     3241 tgctcctcaa gcacggcccc aacagtgaag tagctgattg tcatcagcgc attgacggcg
     3301 tccccggccg aaaaacccgc ctcgcagagg aagcgaagct gcgcgtcggc cgtttccatc
     3361 tgcggtgcgc ccggtcgcgt gccggcatgg atgcgcgcgc catcgcggta ggcgagcagc
     3421 gcctgcctga agctgcgggc attcccagtc agaaatgagc gccagtcgtc gtcggctctc
     3481 ggcaccgaag tgctatgatt ctccgccagc atggcttcgg ccagtgcgtc gagcagcgcc
     3541 cgcttgttcc tgaagtgcca gtaaagcgcc ggctgctgaa cccccaaccg ttccgccagt
     3601 ttgcgtgtcg tcagaccgtc tacgccgacc tcgttcaaca ggtctagggc ggcacggatc
     3661 actgtattcg gctgcaactt tgtcatgctt gacactttat cactgataaa cataatatgt
     3721 ccaccaactt atcagtgata aagaatccgc gcgttcaatc ggaccagcgg aggctggtcc
     3781 ggaggccaga cgtgaaaccc aacatacccc tgatcgtaat tctgagcact gtcgcgctcg
     3841 acgctgtcgg catcggcctg attatgccgg tgctgccggg cctcctgcgc gatctggttc
     3901 actcgaacga cgtcaccgcc cactatggca ttctgctggc gctgtatgcg ttggtgcaat
     3961 ttgcctgcgc acctgtgctg ggcgcgctgt cggatcgttt cgggcggcgg ccaatcttgc
     4021 tcgtctcgct ggccggcgcc actgtcgact acgccatcat ggcgacagcg cctttccttt
     4081 gggttctcta tatcgggcgg atcgtggccg gcatcaccgg ggcgactggg gcggtagccg
     4141 gcgcttatat tgccgatatc actgatggcg atgagcgcgc gcggcacttc ggcttcatga
     4201 gcgcctgttt cgggttcggg atggtcgcgg gacctgtgct cggtgggctg atgggcggtt
     4261 tctcccccca cgctccgttc ttcgccgcgg cagccttgaa cggcctcaat ttcctgacgg
     4321 gctgtttcct tttgccggag tcgcacaaag gcgaacgccg gccgttacgc cgggaggctc
     4381 tcaacccgct cagcttcgtt cggtgggccc ggggcatgac cgtcgtcgcc gccctgatgg
     4441 cggtcttctt catcatgcaa cttgtcggac aggtgccggc cgcgctttgg gtcattttcg
     4501 gcgaggatcg ctttcactgg gacgcgacca cgatcggcat ttcgcttgcc gcatttggca
     4561 ttctgcattc actcgcccag gcaatgatca ccggccctgt agccgcccgg ctcggcgaaa
     4621 ggcgggcact catgctcgga atgattgccg acggcacagg ctacatcctg cttgccttcg
     4681 cgacacgggg atggatggcg ttcccgatca tggtcctgct tgcttcgggt ggcatcggaa
     4741 tgccggcgct gcaagcaatg ttgtccaggc aggtggatga ggaacgccag gggcagctgc
     4801 aaggctcact ggcggcgctc accagcctga cctcgatcgt cggacccctc ctcttcacgg
     4861 cgatctatgc ggcttctata acaacgtgga acgggtgggc atggattgca ggcgctgccc
     4921 tctacttgct ctgcctgccg gcgctgcgtc gcgggctttg gagcggcgca gggcaacgag
     4981 ccgatcgctg atcgtggaaa cgatagggac ggatcgctgc agcgcaaact attaactggc
     5041 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5101 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5161 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5221 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5281 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5341 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5401 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5461 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg ggggatcagg
     5521 accgctgccg gagcgcaacc cactcactac agcagagcca tgtagggccg ccggcgttgt
     5581 ggataccacg cggaaaactt ggccctcact gacagatgag gggcggacgt tgacacttga
     5641 ggggccgact cacccggcgc ggcgttgaca gatgaggggc aggctcgatt tcggccggcg
     5701 acgtggagct ggccagcctc gcaaatcggc gaaaacgcct gattttacgc gagtttccca
     5761 cagatgatgt ggacaagcct ggggataagt gccctgcggt attgacactt gaggggcgcg
     5821 actactgaca gatgaggggc gcgatccttg acacttgagg ggcagagtga tgacagatga
     5881 ggggcgcacc tattgacatt tgaggggctg tccacaggca gaaaatccag catttgcaag
     5941 ggtttccgcc cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt taaaccaata
     6001 tttataaacc ttgtttttaa ccagggctgc gccctggcgc gtgaccgcgc acgccgaagg
     6061 ggggtgcccc cccttctcga accctcccgg cccgctaacg cggcacccca tccccccagg
     6121 ggctgcgccc ctcggccgcg aacgacctca ccccaaaaat ggcagccacg tagaaagcca
     6181 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     6241 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     6301 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     6361 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     6421 gcaggggatc aagatcgacg gatcgatccg gggaattaat tccggggcaa tcccgcaagg
     6481 agggtgaatg aatcggacgt ttgaccggaa ggcatacagg caagaactga tcgacgcggg
     6541 gttttccgcc gaggatgccg aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga
     6601 aaccttccag tccgtcggct cgatggtcca gcaagctacg gccaagatcg agcgcgacag
     6661 cgtgcaactg gctccccctg ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg
     6721 tctcgaacag gaggcggcag gtttggcgaa gtcgatgacc atcgacacgc gaggaactat
     6781 gacgaccaag aagcgaaaaa ccgccggcga ggacctggca aaacaggtca gcgaggccaa
     6841 gcaggccgcg ttgctgaaac acacgaagca gcagatcaag gaaatgcagc tttccttgtt
     6901 cgatattgcg ccgtggccgg acacgatgcg agcgatgcca aacgacacgg cccgctctgc
     6961 cctgttcacc acgcgcaaca agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt
     7021 ccacgtcaac aaggacgtga agatcaccta caccggcgtc gagctgcggg ccgacgatga
     7081 cgaactggtg tggcagcagg tgttggagta cgcgaagcgc acccctatcg gcgagccgat
     7141 caccttcacg ttctacgagc tttgccagga cctgggctgg tcgatcaatg gccggtatta
     7201 cacgaaggcc gaggaatgcc tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga
     7261 ccgcgttggg cacctggaat cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg
     7321 caagaaaacg tcccgttgcc aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg
     7381 cgaccactac acgaaattca tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg
     7441 gatgttcgac tatttcagct cgcaccggga gccgtacccg ctcaagctgg aaaccttccg
     7501 cctcatgtgc ggatcggatt ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc
     7561 ctgcgaagag ttgcgaggca gcggcctggt ggaacacgcc tgggtcaatg atgacctggt
     7621 gcattgcaaa cgctagggcc ttgtggggtc agttccggct gggggttcag cagccacctg
     7681 catcgcaagc tagcttgcta gagggtcagc tttatgcttg taaaccgttt tgtgaaaaaa
     7741 tttttaaaat aaaaaagggg acctctaggg tccccaatta attagtaata taatctatta
     7801 aaggtcattc aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg ctgggtgcca
     7861 agctctcggg taacatcaag gcccgatcct tggtagtcgg caaataatgt ctaacaattc
     7921 gttcaagccg acgccgcttc gcggcgcggc ttaactcaag cgttagatgc actaagcaca
     7981 taattgctca cagccaaact atcaggtcaa gtctgctttt attattttta agcgtgcata
     8041 ataagcccta cacaaattgg gagatatatc atgaaaggct ggctttttct tgttatcgca
     8101 atagttggcg aagtaatcgc aacatccgca ttaaaatcta gcgagggctt tactaagctg
     8161 atccggtgga tgaccttttg aatgaccttt aatagattat attactaatt aattggggac
     8221 cctagaggtc ccctttttta ttttaaaaat tttttcacaa aacggtttac aagcataaag
     8281 ctgactctag ctagag
//