pHT304枯草芽孢杆菌表达载体

基本信息
启动子: Lac
复制子: ColE1 ori
质粒分类: 广宿主系列，枯草杆菌载体
质粒大小: 6530bp
质粒标签: LacZa
原核抗性: Amp
筛选标记: erm
克隆菌株: DH5α
培养条件: 37℃，有氧 LB
表达宿主: 枯草芽孢杆菌
诱导方式: IPTG诱导
5'测序引物: M13R:CAGGAAACAGCTATGACC
3'测序引物: M13F:TGTAAAACGACGGCCAGT
质粒简介
   The promoter of pHT304 vector is Lac, which can be used to induce the expression of recombinant protein in Escherichia coli DE3 engineering bacteria (DE3 BL21 or DE3 Rosetta). The tag has LacZa, which can be purified by nickel column.

质粒图谱

质粒序列
LOCUS       Exported File           6530 bp ds-DNA    circular SYN 29-9-2015
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
 ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
 AUTHORS   .
 TITLE     Direct Submission
 JOURNAL   Exported 2015-9-29 from http://www.biovector.net
FEATURES             Location/Qualifiers
    source          1..6530
                    /organism="synthetic DNA construct"
                    /mol_type="other DNA"
    CDS             3862..4722
                    /codon_start=1
                    /gene="bla"
                    /product="beta-lactamase"
                    /note="AmpR"
                    /note="confers resistance to ampicillin, carbenicillin, and
                    related antibiotics"
                    /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                    ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                    PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                    EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                    LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                    LIKHW"
    rep_origin      4893..5481
                    /direction=RIGHT
                    /note="ori"
                    /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                    replication"
    protein_bind    5769..5790
                    /bound_moiety="E. coli catabolite activator protein"
                    /note="CAP binding site"
                    /note="CAP binding activates transcription in the presence
                    of cAMP."
    promoter        5805..5835
                    /note="lac promoter"
                    /note="promoter for the E. coli lac operon"
    protein_bind    5843..5859
                    /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     5867..5883
                    /note="M13 rev"
                    /note="common sequencing primer, one of multiple similar
                    variants"
    CDS             5879..6202
                    /codon_start=1
                    /gene="lacZ?fragment"
                    /product="LacZ-alpha fragment of beta-galactosidase"
                    /note="lacZ-alpha"
                    /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                    LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                    DAA"
    misc_feature    5896..5952
                    /gene="
                    "
                    /note="MCS"
                    /note="pUC18/19 multiple cloning site"
    primer_bind     complement(5953..5969)
                    /note="M13 fwd"
                    /note="common sequencing primer, one of multiple similar
                    variants"
ORIGIN
       1 ccatcctcca aagttggaga gtgagtttta tgtcgcaaat attaatgttt ctggtgaacc
      61 ttatcaaatt ttcgttgatt taatagaaac atagcggtaa aattagcagt aacttaatag
     121 aacggaaatg aaaaaagcca ctctcatatg ctattggcta ccaaccttta gcgagaatga
     181 cttaatcctg tacagccata caggacttcg acttataaga ggcgccaacc tcaaataagt
     241 tatttgcctt gttttcgcga acaaggctta ttagatacac ctattgtacc gttactctac
     301 gaatatttca actagtaatt actagcattg tcatatacat aataaaacgg atataaaagg
     361 gcgttttcta tacctagaag tcttgtaaat gtacagggcg tttagatata gagaacgccc
     421 tttttgtgtt ccgttccagt ggaggctacc actttaaaaa gatggtctag tgtagccaat
     481 gcaggagagt acactcggat atcagttgtc gttgcattca actgtctgac gtaagcgagg
     541 taaaggacac aagccttgca taaaacaagc ctacgggatg taaatcctaa taatgatgat
     601 aaccaagacg ttagcggcaa aaagtgttgg gggttcaaaa taagacatga ttgtgcgact
     661 ggagttaaac agttactcgt aagcggcgat catgacactg attcacggct attcttgtac
     721 aaactttatt acaaggatat gcgggttata tagcgaatca cccgaaaggg aacggtgttg
     781 ggcgtgagaa acgcaccgta cggcgcaata caatgccaat aagctatata cggacggtat
     841 agtagttttg taagctataa ccgtttgtcg tcaatgcaac caatctcaat tcgagacctc
     901 ggcatctaag ccagtacgaa tgagtgggcg ttttaacctc gtaaattttc aacaggggtt
     961 actatgccca aaactacatt cagatttcct aacaaactcg ccagtatgaa aaccttaaga
    1021 ccttaaagtc aagggatttg aaggatttta acctcgatta gcaaaaaatg tagagtactg
    1081 aagcaactac cattaactaa gatagtgggg gattgaggaa gaatccagag ctgtttaaat
    1141 caagtgaaag acaagatgaa attaaaagaa tagtgaaaga taggggagtg gttctctatg
    1201 agaaaggaaa tggctagaga acaaaggcag cggtttattg atctattgtt agactttatg
    1261 gtaaagaatc ctcatttatt tgttaatggt acagaggatg aaagtaataa tgttgttaca
    1321 aaatgtaata gtgatattaa agaggttgcg gagtcatatt taactctttt atagtgagag
    1381 ggttaaaact aattaatatg tattaaggcc caatgttgga attattgtat ttcactaggc
    1441 aacctactta ctaaaagtaa gattatccat tagtggatgt tataatattg ggttttttaa
    1501 cacaataatc atcgcctttc ggtgtcgttt gatagaaaag taaccattag cgatgaaaaa
    1561 gtcaatataa aaagccatcc gtaaaaaacg gatggcttac cgtacatagg atcgttggta
    1621 gggcggcgta tcctacatct ctggtaactg acctagccaa tcaaatgctt gagaacggcg
    1681 gttagataag cgcgtgggga acctttccca cctcaaagat cctatatcat tattatgtta
    1741 ctttctacag gtagtatacc atgttcttat attttagtaa actccccgtt agcttaacag
    1801 gtctttgtaa gcaattaaac gtccactatt caatcgtctt tggattttcg caggaccgtt
    1861 ttttagatcg aacatagttg ataagaacaa ataaccgctt gggtccaact ttatagcaat
    1921 tagtatatgg tcatttaaaa tctttaccaa ttcaacgcta ttaggttctt taggattttg
    1981 cccgacatag tcggggtgtt caacgatatc ttttatgtgc gatgaatatt tttcataaat
    2041 accaggatgt tgtttcttta cgtgctttat aaatccggga aacattttta catcgttaga
    2101 agtgcaagtc aagttatatg tatctataat gatttgtgga agttttgcca caacagttgg
    2161 tttatttaca atcttttttt tattagccgt caaatttctc cctcatctcg tctctttata
    2221 tctttatttt atcataaagg agtatttgaa ccgtcgcgcg ggacaggttt atgataggga
    2281 tattttattg aataattgat ggtataaggg actttcatgc ttggaaagtg gggattatga
    2341 attagatgct tgtccacaat atgttccaat gtaattaaaa tttatgttcc caccttgacc
    2401 aaacatcacg tccatactta aatcgtccct cctttaatag gtaaaatatt aatttacctt
    2461 aataaaaaaa taatggataa tagtattcgt ctgaatttat ataatcaggg ggaactattg
    2521 atgctgggga tactatttac agcggcgcca tctactgatg tcgtaaagga tttgcaagat
    2581 aaagttatat cattgcagga tcatgaggta gcgtttttga acaccacgat atctaatatg
    2641 ttgatcccct tagaagcaaa cttaagagtg tgttgatagt gcagtatctt aaaattttgt
    2701 gtataatagg aattgaagtt aaattagatg ctaaaaattt gtaattaaga aggagggatt
    2761 cgtcatgttg gtattccaaa tgcgtaatgt agataaaaca tctactgttt tgaaacagac
    2821 taaaaacagt gattacgcag ataaataaat acgttagatt aattcctacc agtgactaat
    2881 cttatgactt tttaaacaga taactaaaat tacaaacaaa tcgtttaact tctgtattta
    2941 tttacagatg taatcacttc aggagtaatt acatgaacaa aaatataaaa tattctcaaa
    3001 actttttaac gagtgaaaaa gtactcaacc aaataataaa acaattgaat ttaaaagaaa
    3061 ccgataccgt ttacgaaatt ggaacaggta aagggcattt aacgacgaaa ctggctaaaa
    3121 taagtaaaca ggtaacgtct attgaattag acagtcatct attcaactta tcgacagaaa
    3181 aattaaaact gaacattcgt gtcactttaa ttcaccaaga tattctacag tttcaattcc
    3241 ctaacaaaca gaggtataaa attgttggga gtattcctta ccatttaagc acacaaatta
    3301 ttaaaaaagt ggtttttgaa agccatgcgt ctgacatcta tctgattgtt gaagaaggat
    3361 tctacaagcg taccttggat attcaccgaa cactagggtt gctcttgcac actcaagtct
    3421 cgattcagca attgcttaag ctgccagcgg aatgctttca tcctaaacca aaagtaaaca
    3481 gtgtcttaat aaaacttacc cgccatacca cagatgttcc agataaatat tggaagctat
    3541 atacgtactt tgtttcaaaa tgggtcaatc gagaatatcg tcaactgttt actaaaaatc
    3601 agtttcatca agcaatgaaa cacgccaaag taaacaattt aagtaccatt acttatgagc
    3661 aagtattgtc tatttttaat agttatctat tatttaacgg gaggaaataa ttctatgagt
    3721 cgctttttta aatttggaaa gttacacgtt actaaaggga atggagataa attattagat
    3781 atactactga cagcttccaa gaaggtaaag aggtccctag cgcctacggg gaatttgtat
    3841 cgggattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
    3901 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
    3961 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
    4021 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
    4081 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
    4141 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
    4201 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
    4261 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
    4321 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
    4381 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
    4441 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
    4501 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
    4561 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
    4621 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
    4681 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
    4741 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
    4801 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
    4861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
    4921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    4981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
    5041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    5101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    5161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    5221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    5281 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    5341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    5401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    5461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    5521 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    5581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    5641 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
    5701 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
    5761 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
    5821 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
    5881 gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc
    5941 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
    6001 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
    6061 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
    6121 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
    6181 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
    6241 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
    6301 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
    6361 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
    6421 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
    6481 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
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