Basic Vector Information
pGEX-4T-2 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-4T-2 vector Sequence
LOCUS pGEX-4T-2. 4970 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial vector for expressing fusion proteins with a thrombin site. For other reading frames, use pGEX-4T-1 or pGEX-4T-3. ACCESSION . VERSION . KEYWORDS pGEX-4T-2. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4970) AUTHORS GE Healthcare TITLE Direct Submission REFERENCE 2 (bases 1 to 4970) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..4970 /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" misc_feature 972..982 /label=stop codons /note="stop codons" /note="stop codons in all three reading frames" promoter 1273..1377 /label=AmpR promoter CDS 1378..2235 /label=AmpR /note="beta-lactamase" rep_origin 2409..2997 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 3241..3318 /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 3319..4398 /label=lacI /note="lac repressor" protein_bind 4414..4435 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 4450..4480 /label=lac promoter /note="promoter for the E. coli lac operon" CDS 4524..4697 /label=lacZ-alpha /note="LacZ-alpha fragment of beta-galactosidase"
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