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
文献和实验
技术资料