pIEFBPR

$260.00

  • Model: PVT11306
  • 50 Units in Stock
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pIEFBPR

Catalog No. PVT11306
Packing 2ug

 

pIEFBPR Information

Replicon: ColE1 Ori

Plasmid classification: Bacillus subtilis plasmid; hay integration plasmid; hay secretory plasmid.

Plasmid size: 8965bp

Prokaryotic resistance: ampicillin Amp

Cloned strains of Escherichia coli, DH5 A and other Escherichia coli

Culture conditions: 37 centigrade, aerobic, LB

 

pIEFBPR Description

pIEFBPR plasmid is a subtype integrated secretion vector.

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.

 

pIEFBPR Reference

mazF, a novel counter-selectable marker for unmarked chromosomal manipulation in Bacillus subtilis

 

pIEFBPR Sequence

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  
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
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