Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010916 | pGEX-4T-3 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pGEX-4T-3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4968 bp
- Type:
- pGEX Vectors (GE Healthcare)
- Replication origin:
- ori
- Source/Author:
- GE Healthcare
- Copy Number:
- High copy number
pGEX-4T-3 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pGEX-4T-3 vector Sequence
LOCUS pGEX-4T-3. 4968 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-2.
ACCESSION .
VERSION .
KEYWORDS pGEX-4T-3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4968)
AUTHORS GE Healthcare
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4968)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4968
/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 970..980
/label=stop codons
/note="stop codons"
/note="stop codons in all three reading frames"
promoter 1271..1375
/label=AmpR promoter
CDS 1376..2233
/label=AmpR
/note="beta-lactamase"
rep_origin 2407..2995
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3239..3316
/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 3317..4396
/label=lacI
/note="lac repressor"
protein_bind 4412..4433
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4448..4478
/label=lac promoter
/note="promoter for the E. coli lac operon"
CDS 4522..4695
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"