Basic Vector Information
- Vector Name:
- pET-52b(+)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5227 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
pET-52b(+) 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 ℃.
pET-52b(+) vector Sequence
LOCUS pET-52b(+). 5227 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector for expressing proteins with a cleavable N-terminal
Strep-Tag II tag and a cleavable C-terminal 10xHis tag.
ACCESSION .
VERSION .
KEYWORDS pET-52b(+).
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5227)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5227)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5227
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(26..73)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(113..139)
/label=9xHis
/note="9xHis affinity tag"
CDS complement(146..163)
/label=thrombin site
/note="thrombin recognition and cleavage site"
misc_feature 167..208
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS complement(209..232)
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
CDS complement(239..262)
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
CDS complement(269..271)
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
RBS complement(278..300)
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(315..339)
/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(340..358)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 671..748
/label=lacI promoter
CDS 749..1828
/label=lacI
/note="lac repressor"
protein_bind 1844..1865
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 2403..2591
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 2696..2838
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3024..3612)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3693..3785
/label=AmpR promoter
CDS 3786..4643
/label=AmpR
/note="beta-lactamase"
rep_origin complement(4761..5216)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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