pBBR1MCS2-Tac-mCherry广宿主表达质粒

基本信息

启动子:	Tac
复制子:	pBBR1 Rep，pBBR1 oriV
终止子:	T7 terminator
质粒分类:	红色荧光蛋白表达质粒；广宿主质粒
质粒大小:	6195bp
质粒标签:	mCherry
原核抗性:	卡那霉素Kanamycin
筛选标记:	DH5α
克隆菌株:	DH5α
培养条件:	37℃，有氧，LB
表达宿主:	革兰氏阴性菌
5'测序引物:	M13R:CAGGAAACAGCTATGACC
3'测序引物:	M13F:TGTAAAACGACGGCCAGT

质粒简介

Tac强启动子启动mCherry红色荧光蛋白的表达。

质粒图谱

质粒序列

LOCUS       Exported                6195 bp ds-DNA     circular SYN 11-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBBR1MCS2-Tac-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6195)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, September 11, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..6195
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             458..1252
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1657..1678
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1693..1723
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1755..1771
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1792..1810
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1840..1856
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1910..1987)
                     /gene="lacI"
                     /note="lacI promoter"
     misc_feature    2033..2061
                     /note="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             2108..2818
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /note="mCherry"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      2852..2899
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2941..2957)
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2990..3008)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3018..3034)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3741..4403)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /note="pBBR1 Rep"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      4404..5173
                     /note="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
      241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
      301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
      361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
     1561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1801 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
     1861 gcttgatatc gaattcgagc tcggtacctc gcgaatgcat ctagatcaca ttcaccaccc
     1921 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     1981 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa atttgacaat
     2041 taatcatcgg ctcgtataat gtgtggaatg tctcatgagc ggatatttca cacaggaaac
     2101 agtattcatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2161 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2221 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2281 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2341 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2401 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2461 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2521 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2581 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2641 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2701 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2761 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaatt
     2821 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     2881 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacacatcg
     2941 gatccactag ttctagagcg gccgccaccg cggtggagct ccaattcgcc ctatagtgag
     3001 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3061 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3121 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaaattg
     3181 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3241 accaataggc cgactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
     3301 ggtgccctta aacgcctggt gctacgcctg aataagtgat aataagcgga tgaatggcag
     3361 aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta aatagccgct
     3421 tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgac cgtgtgcttc
     3481 tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat tgacgatatg
     3541 atcatttatt ctgcctccca gagcctgata aaaacggtga atccgttagc gaggtgccgc
     3601 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     3661 agacaaggta tagggcggcg aggcggctac agccgatagt ctggaacagc gcacttacgg
     3721 gttgctgcgc aacccaagtg ctaccggcgc ggcagcgtga cccgtgtcgg cggctccaac
     3781 ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc
     3841 cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg
     3901 ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc cggatacagg
     3961 gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca
     4021 accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg gccccacgcc
     4081 acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt cacgacggag
     4141 atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa agaacgtccg
     4201 atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc catctgcgcc
     4261 acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc ggcccacgcc
     4321 tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc ttgaatgccg
     4381 atttctctgg actgcgtggc catgcttatc tccatgcggt agggtgccgc acggttgcgg
     4441 caccatgcgc aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa
     4501 agtggcagtc aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg
     4561 caatgagctg ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc
     4621 gccctatatc tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc
     4681 ttcggcgcgg ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg
     4741 cggccttcgg ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt
     4801 gaggctcggg gggcaggcgg gcgggcttcg ccttcgactg cccccactcg cataggcttg
     4861 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg
     4921 ccttccactt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc
     4981 cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt
     5041 gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca
     5101 ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccgagc gaagcgagcc
     5161 agccggtggc cgctcgcggc catcgtccac atatccacgg gctggcaagg gagcgcagcg
     5221 accgcgcagg gcgaagcccg gagagcaagc ccgtagggcg ccgcagccgc cgtaggcggt
     5281 cacgactttg cgaagcaaag tctagtgagt atactcaagc attgagtggc ccgccggagg
     5341 caccgccttg cgctgccccc gtcgagccgg ttggacacca aaagggaggg gcaggcatgg
     5401 cggcatacgc gatcatgcga tgcaagaagc tggcgaaaat gggcaacgtg gcggccagtc
     5461 tcaagcacgc ctaccgcgag cgcgagacgc ccaacgctga cgccagcagg acgccagaga
     5521 acgagcactg ggcggccagc agcaccgatg aagcgatggg ccgactgcgc gagttgctgc
     5581 cagagaagcg gcgcaaggac gctgtgttgg cggtcgagta cgtcatgacg gccagcccgg
     5641 aatggtggaa gtcggccagc caagaacagc aggcggcgtt cttcgagaag gcgcacaagt
     5701 ggctggcgga caagtacggg gcggatcgca tcgtgacggc cagcatccac cgtgacgaaa
     5761 ccagcccgca catgaccgcg ttcgtggtgc cgctgacgca ggacggcagg ctgtcggcca
     5821 aggagttcat cggcaacaaa gcgcagatga cccgcgacca gaccacgttt gcggccgctg
     5881 tggccgatct agggctgcaa cggggcatcg agggcagcaa ggcacgtcac acgcgcattc
     5941 aggcgttcta cgaggccctg gagcggccac cagtgggcca cgtcaccatc agcccgcaag
     6001 cggtcgagcc acgcgcctat gcaccgcagg gattggccga aaagctggga atctcaaagc
     6061 gcgttgagac gccggaagcc gtggccgacc ggctgacaaa agcggttcgg caggggtatg
     6121 agcctgccct acaggccgcc gcaggagcgc gtgagatgcg caagaaggcc gatcaagccc
     6181 aagagacggc ccgag
//