Basic Vector Information
- Vector Name:
- pGEX-2TK
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4969 bp
- Type:
- pGEX Vectors (GE Healthcare)
- Replication origin:
- ori
- Source/Author:
- GE Healthcare
- Copy Number:
- High copy number
pGEX-2TK 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 ℃.
pGEX-2TK vector Sequence
LOCUS pGEX-2TK. 4969 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector for expressing GST fusions with a thrombin site and
a cAMP kinase site.
ACCESSION .
VERSION .
KEYWORDS pGEX-2TK.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4969)
AUTHORS GE Healthcare
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4969)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4969
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 183..211
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 219..235
/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)."
CDS 258..911
/label=GST
/note="glutathione S-transferase from Schistosoma
japonicum"
CDS 918..935
/label=thrombin site
/note="thrombin recognition and cleavage site"
CDS 936..950
/codon_start=1
/product="cAMP-dependent protein kinase recognition site"
/label=cAMP-dependent protein kinase recognition site
/note="kinase site"
/translation="RRASV"
misc_feature 951..966
/label=MCS
/note="MCS"
/note="multiple cloning site"
misc_feature 971..981
/label=stop codons
/note="stop codons"
/note="stop codons in all three reading frames"
promoter 1272..1376
/label=AmpR promoter
CDS 1377..2234
/label=AmpR
/note="beta-lactamase"
rep_origin 2408..2996
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3240..3317
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 3318..4397
/label=lacI
/note="lac repressor"
protein_bind 4413..4434
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4449..4479
/label=lac promoter
/note="promoter for the E. coli lac operon"
CDS 4523..4696
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
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