• 规格：20μl质粒
• 复制子：ColE1 Ori
• 质粒分类：枯草杆菌质粒；枯草整合质粒；枯草分泌质粒
• 质粒大小：8965bp
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：DH5α等大肠杆菌
• 质粒简介：            pIEFBPR质粒是一个枯草整合型分泌载体，该质粒的参考文献：mazF, a novel counter-selectable marker for unmarked chromosomal manipulation in Bacillus subtilis            ColE1 ori, ColE1 replication origin; amp, ampicillin-resistance marker; spc, spectinomycin-resistance marker; lacI, lac-repressor-encoding gene; Pspac, Pspac promoter; mazF, E.coli toxin gene; Plpp, promoter of E.coli lpp gene; mazE, E.coli antitoxin gene; DR, directly-repeated sequence; amyE-front and amyE-back, upstream and downstream sequences of the B.subtilis amyE gene, respectively; bpr-front and bpr-back, upstream and downstream sequences of the B.subtilis bpr gene, respectively. All of the DNA sequences located between the amyE-front and amyE-back or the bpr-front and bpr-back allow integration into the chromosome of B.subtilis via a double-crossover event, resulting in selection for spectinomycin-resistance. The unique restriction enzyme sites are highlighted in blue.
• 质粒图谱1： png/1-1F9151G53C07.png

质粒序列： 点击查看序列

LOCUS       Exported                8965 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 8965)

  TITLE     Direct Submission

  JOURNAL   Exported Sep 15, 2017 from MiaoLingPlasmid

            http://www.fenghbio.cn

FEATURES             Location/Qualifiers

     source          1..8965

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      4..459

                     /direction=RIGHT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        626..644

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    692..1697

                     /label=bpr-front

                     /note="upstream of Bacillus subtilis bpr gene"

     misc_feature    1728..1852

                     /label=DR sequence

     CDS             complement(1900..2982)

                     /codon_start=1

                     /gene="lacI"

                     /product="lac repressor"

                     /label=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"

     CDS             complement(3232..3567)

                     /codon_start=1

                     /label=mazF

                     /note="Escherichia coli toxin MazF"

                     /translation="MVSRYVPDMGDLIWVDFDPTKGSEQAGHRPAVVLSPFMYNNKTGM

                     CLCVPCTTQSKGYPFEVVLSGQERDGVALADQVKSIAWRARGATKKGTVAPEELQLIKA

                     KINVLIG"

     CDS             4179..4931

                     /codon_start=1

                     /product="spectinomycin adenyltransferase; selectable 

                     either in Escherichia coli or Bacillus subtilis"

                     /label=spc

                     /translation="LNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL

                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG

                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD

                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR

                     SYLGENIEWTNENVNLTINYLNNRLKKL"

     misc_feature    4933..5057

                     /label=DR sequence

     misc_feature    5197..6235

                     /label=bpr-back

                     /note="downstream of Bacillus subtilis bpr gene; homologous

                     arm for recombination"

     misc_feature    6450..6758

                     /label=mazE

     CDS             complement(7218..7806)

                     /codon_start=1

                     /label=ColE1 ori

                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 

                     replication"

                     /translation="LRSFFSARNLLLANKKTTATSGGLFAGSRATNSFSEGNWLQQSAD

                     TKYCPSSVAVVRPPLQELCSTAYIPRSANPVTSGCCQWR*VVSYRVGLKTIVTG*GAAV

                     GLNGGFVHTAQLGANDLHRTEIPTA*AMRKRHASRREKGGQVSGKRQGRNRRAHEGASR

                     GKRLVSL*SCRVSPPLT*ASIFVMLVRGAEPME"

     CDS             complement(7977..8837)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(8838..8942)

                     /gene="bla"

                     /label=AmpR promoter

ORIGIN

        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga

       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg

      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat

      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg

      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata

      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt

      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat

      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg

      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg

      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg

      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg

      661 gccccccctc gagttaacct gcaggcgcgc ctttgatggg tatcggtatt gctactgggg

      721 aaaatcgcgc ggcagaggca gcaaaaaaag caatttccag cccgcttctt gaagcggcca

      781 ttgacggtgc gcaaggcgtc ctcatgaaca tcactggagg aacaaacctc agcctatatg

      841 aggttcagga agcagcagac attgtcgctt cggcgtctga tcaagacgta aacatgattt

      901 tcggttctgt tattaatgaa aatctaaaag atgagattgt ggtgacagtg attgcaaccg

      961 gctttatcga acaagagaag gacgtgacga agcctcagcg cccaagctta aatcaaagca

     1021 tcaaaacaca caatcaaagt gttccgaagc gtgacgcaaa acgtgaggaa cctcagcagc

     1081 agaacacagt aagccgtcat acttcacagc cggctgatga tacgcttgac atcccgacat

     1141 tcttaagaaa ccgtaataaa cgcggctaat gtaaaggaca aaatcgtttt cgattttgtc

     1201 ttttttgttt ttctcttcac acttccttct tataaagtct ttttccctat tgctttcctt

     1261 cgcttagtaa caaaacagat aattagaccc atttattttt gtgacatttt tatcattttc

     1321 atatatatgg aaattgaatg acatgaaacg acaatatctg taattcagat tgtctacagt

     1381 taatatacag cgatgttctg acaaaccatt cattattaaa aggagggacg acactttttt

     1441 taaaaagcat gttgaaaaag ggggatgaaa atgaggaaaa aaacgaaaaa cagactcatc

     1501 agctctgttt taagtacagt tgtcatcagt tcactgctgt ttccgggagc agccggggca

     1561 agcagtaaag tcacctcacc ttctgttaaa aaggagcttc aatctgcgga atccattcaa

     1621 aacaagattt cgagttcatt aaagaaaagc tttaaaaaga aagaaaaaac gacttttctg

     1681 attaaattta aagatctatc gatgctagcc atggtcgacg cggccgcaaa attgaaaaaa

     1741 tggtggaaac acttttttca atttttttgt tttattattt aatatttggg aaatattcat

     1801 tctaattggt aatcagattt tagaaaacaa taaacccttg cataggggga tctccggagt

     1861 ttaaaccgcg gatcctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg

     1921 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg

     1981 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc

     2041 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag

     2101 gcgaaaatcc tgtttgatgg tggttgacgg cgggatataa catgagctgt cttcggtatc

     2161 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg

     2221 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc

     2281 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc

     2341 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg

     2401 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac

     2461 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg

     2521 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc

     2581 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag

     2641 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac

     2701 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc

     2761 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg

     2821 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg

     2881 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac

     2941 accggcatac tctgcgacat cgtataacgt tactggtttc atcaaaatcg tctccctccg

     3001 tttgaatatt tgattgatcg taaccagatg aagcactctt tccactatcc ctacagtgtt

     3061 atggcttgaa caatcacgaa acaataattg gtacgtacga tctttcagcc gactcaaaca

     3121 tcaaatctta caaatgtagt ctttgaaagt attacatatg taagatttaa atgcaaccgt

     3181 tttttcggaa ggaaatgatg acctcgtttc caccggaatt agcttgcatg cctacccaat

     3241 cagtacgtta attttggctt taatgagttg taattcctct ggggcaactg ttcctttctt

     3301 cgttgctcct cttgcccgcc aggcgatact ttttacctga tcagctaacg ctacgccatc

     3361 acgttcctga ccggataaaa caacttcgaa cggatatcct tttgattgcg ttgtacaagg

     3421 aacacacaga cacatacctg ttttgttgtt gtacatgaaa ggactcagga caacagctgg

     3481 acgatgtcca gcttgctcgc taccttttgt cgggtcaaaa tcaacccaaa tcagatcgcc

     3541 catatcgggt acgtatcggc ttaccatacc tgcttcctcc ttaagcttaa ttgttatccg

     3601 ctcacaattc cacacattat gccacacctt gtagataaag tcaacaactt tttgcaaaat

     3661 ttttcaggaa ttttagcaga ggttgttctg gatgtagaac aaaacatctt t