Basic Vector Information
- Vector Name:
- pCOLD-MazF-K56A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6090 bp
- Type:
- pCOLD Vectors
- Replication origin:
- ori
- Source/Author:
- Zhang Z, Chen LQ, Zhao YL, et al.
- Copy Number:
- High copy number
- Promoter:
- cspA
- Cloning Method:
- Enzyme Cut
- Fusion Tag:
- Trigger Factor (ribosome-associated chaperone from E. coli (Hoffmann et al., 2010))
- Expression Method:
- IPTG induced
pCOLD-MazF-K56A vector Vector Map
Plasmid Resuspension 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 ℃.
pCOLD-MazF-K56A vector Sequence
LOCUS 40924_12645 6090 bp DNA circular SYN 25-JUL-2018
DEFINITION Vector for cold shock-induced expression in E. coli of a protein
fused to the Trigger Factor chaperone.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6090)
AUTHORS TaKaRa
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6090)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6090
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 15..81
/label=cspA promoter
/note="promoter of the E. coli cold shock protein cspA gene
(Mitta et al., 1997)"
protein_bind 84..100
/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)."
5'UTR 122..255
/label=cspA 5'UTR
/note="5'UTR of the E. coli cold shock protein cspA gene
(Mitta et al., 1997)"
CDS 256..270
/label=TEE
/note="translation enhancing element for E. coli (Qing et
al., 2004)"
CDS 271..288
/label=6xHis
/note="6xHis affinity tag"
CDS 289..1584
/label=Trigger Factor
/note="ribosome-associated chaperone from E. coli (Hoffmann
et al., 2010)"
CDS 1594..1617
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
CDS 1627..1644
/label=thrombin site
/note="thrombin recognition and cleavage site"
CDS 1651..1662
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
CDS 1666..1998
/gene="mazF"
/label=mazF
/note="Endoribonuclease toxin MazF from Escherichia coli
(strain K12). Accession#: P0AE70"
3'UTR 2051..2195
/label=cspA 3'UTR
/note="3'UTR of the E. coli cold shock protein cspA gene
(Mitta et al., 1997)"
rep_origin complement(2394..2849)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2949..3053
/label=AmpR promoter
CDS 3054..3911
/label=AmpR
/note="beta-lactamase"
rep_origin 4085..4673
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind complement(4811..4832)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(4848..5927)
/label=lacI
/note="lac repressor"
promoter complement(5928..6005)
/label=lacI promoter
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