Basic Vector Information
pGEX-5X-1 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-5X-1 vector Sequence
LOCUS pGEX-5X-1. 4972 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial vector for expressing GST fusion proteins with a Factor Xa site. For other reading frames, use pGEX-5X-2 or pGEX-5X-3. ACCESSION . VERSION . KEYWORDS pGEX-5X-1. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4972) AUTHORS GE Healthcare TITLE Direct Submission REFERENCE 2 (bases 1 to 4972) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..4972 /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 921..932 /label=Factor Xa site /note="Factor Xa recognition and cleavage site" misc_feature 934..969 /label=MCS /note="MCS" /note="multiple cloning site" misc_feature 974..984 /label=stop codons /note="stop codons" /note="stop codons in all three reading frames" promoter 1275..1379 /label=AmpR promoter CDS 1380..2237 /label=AmpR /note="beta-lactamase" rep_origin 2411..2999 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 3243..3320 /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 3321..4400 /label=lacI /note="lac repressor" protein_bind 4416..4437 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 4452..4482 /label=lac promoter /note="promoter for the E. coli lac operon" CDS 4526..4699 /label=lacZ-alpha /note="LacZ-alpha fragment of beta-galactosidase"
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