pHT43-His枯草分泌质粒

基本信息

启动子:	Lac
复制子:	ori
质粒分类:	广宿主系列，枯草杆菌载体
质粒大小:	8101bp
原核抗性:	Amp
筛选标记:	Chl
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:	M13F:TGTAAAACGACGGCCAGT
3'测序引物:	根据序列设计引物

质粒简介

    PHT43-His is Bacillus subtilis, the size of 8101bp, which is transformed from the pHT43 vector, screening marks for chloramphenicol.

质粒图谱

质粒序列

LOCUS       Exported                8101 bp ds-DNA     circular SYN 12-JUL-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pHT43-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, September 9, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..8101
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1238..1254
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1297..2379)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2763..2787
                     /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)."
     misc_feature    2809..2901
                     /note="SP amyQ"
     CDS             2959..2976
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     promoter        6178..6282
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             6283..7143
                     /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      7314..7902
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttaagttatt ggtatgactg gttttaagcg caaaaaaagt tgctttttcg tacctattaa
       61 tgtatcgttt tagaaaaccg actgtaaaaa gtacagtcgg cattatctca tattataaaa
      121 gccagtcatt aggcctatct gacaattcct gaatagagtt cataaacaat cctgcatgat
      181 aaccatcaca aacagaatga tgtacctgta aagatagcgg taaatatatt gaattacctt
      241 tattaatgaa ttttcctgct gtaataatgg gtagaaggta attactatta ttattgatat
      301 ttaagttaaa cccagtaaat gaagtccatg gaataataga aagagaaaaa gcattttcag
      361 gtataggtgt tttgggaaac aatttccccg aaccattata tttctctaca tcagaaaggt
      421 ataaatcata aaactctttg aagtcattct ttacaggagt ccaaatacca gagaatgttt
      481 tagatacacc atcaaaaatt gtataaagtg gctctaactt atcccaataa cctaactctc
      541 cgtcgctatt gtaaccagtt ctaaaagctg tatttgagtt tatcaccctt gtcactaaga
      601 aaataaatgc agggtaaaat ttatatcctt cttgttttat gtttcggtat aaaacactaa
      661 tatcaatttc tgtggttata ctaaaagtcg tttgttggtt caaataatga ttaaatatct
      721 cttttctctt ccaattgtct aaatcaattt tattaaagtt catttgatat gcctcctaaa
      781 tttttatcta aagtgaattt aggaggctta cttgtctgct ttcttcatta gaatcaatcc
      841 ttttttaaaa gtcaatatta ctgtaacata aatatatatt ttaaaaatat cccactttat
      901 ccaattttcg tttgttgaac taatgggtgc tttagttgaa gaataaagac cacattaaaa
      961 aatgtggtct tttgtgtttt tttaaaggat ttgagcgtag cgaaaaatcc ttttctttct
     1021 tatcttgata ataagggtaa ctattgccga tcgtccattc cgacagcatc gccagtcact
     1081 atggcgtgct gctagcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1141 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1201 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc
     1261 gagctcaggc cttaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1321 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1381 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     1441 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     1501 aaaatcctgt ttgatggtgg ttgacggcgg gatataacat gagctgtctt cggtatcgtc
     1561 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1621 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1681 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1741 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1801 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1861 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1921 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1981 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     2041 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     2101 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     2161 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     2221 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     2281 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     2341 ggcatactct gcgacatcgt ataacgttac tggtttcatc aaaatcgtct ccctccgttt
     2401 gaatatttga ttgatcgtaa ccagatgaag cactctttcc actatcccta cagtgttatg
     2461 gcttgaacaa tcacgaaaca ataattggta cgtacgatct ttcagccgac tcaaacatca
     2521 aatcttacaa atgtagtctt tgaaagtatt acatatgtaa gatttaaatg caaccgtttt
     2581 ttcggaagga aatgatgacc tcgtttccac cggaattagc ttggtaccag ctattgtaac
     2641 ataatcggta cgggggtgaa aaagctaacg gaaaagggag cggaaaagaa tgatgtaagc
     2701 gtgaaaaatt ttttatctta tcacttgaaa ttggaaggga gattctttat tataagaatt
     2761 gtggaattgt gagcggataa caattcccaa ttaaaggagg aaggatcaat gattcaaaaa
     2821 cgaaagcgga cagtttcgtt cagacttgtg cttatgtgca cgctgttatt tgtcagtttg
     2881 ccgattacaa aaacatcagc cgtaggatcc atgcggccgc taactagtgt caccggtatc
     2941 tagagtcgac gtcccgggca tcaccatcat catcaccagc ccgcctaatg agcgggcttt
     3001 tttcacgtca cgcgtccatg gagatctttg tctgcaactg aaaagtttat accttacctg
     3061 gaacaaatgg ttgaaacata cgaggctaat atcggcttat taggaatagt ccctgtacta
     3121 ataaaatcag gtggatcagt tgatcagtat attttggacg aagctcggaa agaatttgga
     3181 gatgacttgc ttaattccac aattaaatta agggaaagaa taaagcgatt tgatgttcaa
     3241 ggaatcacgg aagaagatac tcatgataaa gaagctctaa aactattcaa taaccttaca
     3301 atggaattga tcgaaagggt ggaaggttaa tggtacgaaa attaggggat ctacctagaa
     3361 agccacaagg cgataggtca agcttaaaga acccttacat ggatcttaca gattctgaaa
     3421 gtaaagaaac aacagaggtt aaacaaacag aaccaaaaag aaaaaaagca ttgttgaaaa
     3481 caatgaaagt tgatgtttca atccataata agattaaatc gctgcacgaa attctggcag
     3541 catccgaagg gaattcatat tacttagagg atactattga gagagctatt gataagatgg
     3601 ttgagacatt acctgagagc caaaaaactt tttatgaata tgaattaaaa aaaagaacca
     3661 acaaaggctg agacagactc caaacgagtc tgttttttta aaaaaaatat taggagcatt
     3721 gaatatatat tagagaatta agaaagacat gggaataaaa atattttaaa tccagtaaaa
     3781 atatgataag attatttcag aatatgaaga actctgtttg tttttgatga aaaaacaaac
     3841 aaaaaaaatc cacctaacgg aatctcaatt taactaacag cggccaaact gagaagttaa
     3901 atttgagaag gggaaaaggc ggatttatac ttgtatttaa ctatctccat tttaacattt
     3961 tattaaaccc catacaagtg aaaatcctct tttacactgt tcctttaggt gatcgcggag
     4021 ggacattatg agtgaagtaa acctaaaagg aaatacagat gaattagtgt attatcgaca
     4081 gcaaaccact ggaaataaaa tcgccaggaa gagaatcaaa aaagggaaag aagaagttta
     4141 ttatgttgct gaaacggaag agaagatatg gacagaagag caaataaaaa acttttcttt
     4201 agacaaattt ggtacgcata taccttacat agaaggtcat tatacaatct taaataatta
     4261 cttctttgat ttttggggct attttttagg tgctgaagga attgcgctct atgctcacct
     4321 aactcgttat gcatacggca gcaaagactt ttgctttcct agtctacaaa caatcgctaa
     4381 aaaaatggac aagactcctg ttacagttag aggctacttg aaactgcttg aaaggtacgg
     4441 ttttatttgg aaggtaaacg tccgtaataa aaccaaggat aacacagagg aatccccgat
     4501 ttttaagatt agacgtaagg ttcctttgct ttcagaagaa cttttaaatg gaaaccctaa
     4561 tattgaaatt ccagatgacg aggaagcaca tgtaaagaag gctttaaaaa aggaaaaaga
     4621 gggtcttcca aaggttttga aaaaagagca cgatgaattt gttaaaaaaa tgatggatga
     4681 gtcagaaaca attaatattc cagaggcctt acaatatgac acaatgtatg aagatatact
     4741 cagtaaagga gaaattcgaa aagaaatcaa aaaacaaata cctaatccta caacatcttt
     4801 tgagagtata tcaatgacaa ctgaagagga aaaagtcgac agtactttaa aaagcgaaat
     4861 gcaaaatcgt gtctctaagc cttcttttga tacctggttt aaaaacacta agatcaaaat
     4921 tgaaaataaa aattgtttat tacttgtacc gagtgaattt gcatttgaat ggattaagaa
     4981 aagatattta gaaacaatta aaacagtcct tgaagaagct ggatatgttt tcgaaaaaat
     5041 cgaactaaga aaagtgcaat aaactgctga agtatttcag cagttttttt tatttagaaa
     5101 tagtgaaaaa aatataatca gggaggtatc aatatttaat gagtactgat ttaaatttat
     5161 ttagactgga attaataatt aacacgtaga ctaattaaaa tttaatgagg gataaagagg
     5221 atacaaaaat attaatttca atccctatta aattttaaca agggggggat taaaatttaa
     5281 ttagaggttt atccacaaga aaagacccta ataaaatttt tactagggtt ataacactga
     5341 ttaatttctt aatgggggag ggattaaaat ttaatgacaa agaaaacaat cttttaagaa
     5401 aagcttttaa aagataataa taaaaagagc tttgcgatta agcaaaactc tttacttttt
     5461 cattgacatt atcaaattca tcgatttcaa attgttgttg tatcataaag ttaattctgt
     5521 tttgcacaac cttttcagga atataaaaca catctgaggc ttgttttata aactcagggt
     5581 cgctaaagtc aatgtaacgt agcatatgat atggtatagc ttccacccaa gttagccttt
     5641 ctgcttcttc tgaatgtttt tcatatactt ccatgggtat ctctaaatga ttttcctcat
     5701 gtagcaaggt atgagcaaaa agtttatgga attgatagtt cctctctttt tcttcaactt
     5761 ttttatctaa aacaaacact ttaacatctg agtcaatgta agcataagat gtttttccag
     5821 tcataatttc aatcccaaat cttttagaca gaaattctgg acgtaaatct tttggtgaaa
     5881 gaattttttt atgtagcaat atatccgata cagcaccttc taaaagcgtt ggtgaatagg
     5941 gcattttacc tatctcctct cattttgtgg aataaaaata gtcatattcg tccatctacc
     6001 tatcctatta tcgaacagtt gaacttttta atcaaggatc agtccttttt ttcattattc
     6061 ttaaactgtg ctcttaactt taacaactcg atttgttttt ccagatctcg agggtaacta
     6121 gcctcgccga tcccgcaaga ggcccggcag tcaggtggca cttttcgggg aaatgtgcgc
     6181 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6241 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6301 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6361 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6421 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6481 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6541 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6601 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6661 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6721 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6781 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6841 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6901 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6961 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7021 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7081 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7141 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7201 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7261 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7321 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7381 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7441 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     7501 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7561 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7621 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7681 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7741 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7801 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7861 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7921 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7981 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8041 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcatg
     8101 c