Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005452 | pIEFBPR | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pIEFBPR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8965 bp
- Type:
- Delivery vector
- Replication origin:
- ori
- Source/Author:
- Zhang XZ, Yan X, Cui ZL, Hong Q, Li SP.
pIEFBPR vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pIEFBPR vector Sequence
LOCUS V005452 8965 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V005452
VERSION V005452
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8965)
AUTHORS Zhang XZ, Yan X, Cui ZL, Hong Q, Li SP.
TITLE mazF, a novel counter-selectable marker for unmarked chromosomal
manipulation in Bacillus subtilis
JOURNAL Nucleic Acids Res. 34 (9), E71 (2006)
PUBMED 16714443
REFERENCE 2 (bases 1 to 8965)
AUTHORS Zhang X-Z., Yan X, Li S-P.
TITLE Direct Submission
JOURNAL Submitted (21-MAR-2006) Department of Microbiology, College of Life
Sciences, Nanjing Agricultural University, Key Laboratory for
Microbiological Engineering of Agricultural Environment, Ministry of
Agriculture, 6 Tongwei Road, Nanjing, Jiangsu 210095, China
REFERENCE 3 (bases 1 to 8965)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8965)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2006"; volume: "34"; issue: "9"; pages: "E71"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-MAR-2006) Department of Microbiology, College of Life Sciences,
Nanjing Agricultural University, Key Laboratory for Microbiological
Engineering of Agricultural Environment, Ministry of Agriculture, 6
Tongwei Road, Nanjing, Jiangsu 210095, China"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8965
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 4..459
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 692..1697
/note="bpr-front; upstream of Bacillus subtilis bpr gene;
homologous arm for recombination"
repeat_region 1728..1852
/label="DR sequence"
/note="DR sequence"
CDS complement(1903..2982)
/label="lacI"
/note="lac repressor"
CDS complement(3235..3567)
/gene="mazF"
/label="Endoribonuclease toxin MazF"
/note="Endoribonuclease toxin MazF from Escherichia coli
(strain K12). Accession#: P0AE70"
protein_bind 3591..3607
/label="lac operator"
/bound_moiety="lac repressor encoded by 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)."
CDS 4179..4931
/codon_start=1
/gene="spc"
/product="Spc"
/label="spc"
/note="spectinomycin adenyltransferase; selectable either
in Escherichia coli or Bacillus subtilis"
/protein_id="ABF31010.1"
/translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
SYLGENIEWTNENVNLTINYLNNRLKKL"
gene 4179..4931
/gene="spc"
/label="spc"
repeat_region 4933..5057
/label="DR sequence"
/note="DR sequence"
misc_feature 5197..6235
/note="bpr-back; downstream of Bacillus subtilis bpr gene;
homologous arm for recombination"
CDS 6510..6755
/gene="mazE"
/label="Antitoxin MazE"
/note="Antitoxin MazE from Escherichia coli (strain K12).
Accession#: P0AE72"
promoter complement(6777..6795)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6816..6832)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6840..6856)
/label="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)."
promoter complement(6864..6894)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(6909..6930)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7218..7806)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7980..8837)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8838..8942)
/label="AmpR promoter"