Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V012941 | pG-KJE8 | 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:
- pG-KJE8
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 11195 bp
- Type:
- Packaging assistance
- Replication origin:
- p15A ori
- Host:
- E. coli
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pG-KJE8 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.
pG-KJE8 vector Sequence
LOCUS V012941 11195 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V012941 VERSION V012941 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 11195) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..11195 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 1789..3702 /gene="dnaK" /label="Chaperone protein DnaK" /note="Chaperone protein DnaK from Escherichia coli O1:K1 / APEC. Accession#: A1A766" CDS 3794..4921 /gene="dnaJ" /label="Chaperone protein DnaJ" /note="Chaperone protein DnaJ from Escherichia coli (strain K12). Accession#: P08622" CDS 4941..5531 /gene="grpE" /label="Protein GrpE" /note="Protein GrpE from Escherichia coli (strain K12). Accession#: P09372" terminator 5837..5923 /label="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6015..6042 /label="rrnB T2 terminator" /note="transcription terminator T2 from the E. coli rrnB gene" promoter 6062..6153 /label="AmpR promoter" rep_origin 6479..7023 /label="p15A ori" /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin." CDS complement(7376..7999) /label="TetR" /note="tetracycline repressor TetR" promoter 8015..8070 /label="tetR/tetA promoters" /note="overlapping promoters for bacterial tetR and tetA" CDS 8386..8439 /label="S loop" /note="GroES chaperone mobile loop that interacts with GroEL" CDS 8678..10321 /gene="groEL" /label="Chaperonin GroEL" /note="Chaperonin GroEL from Escherichia coli O8 (strain IAI1). Accession#: B7M8Q4" terminator 10452..10538 /label="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" promoter 10874..10976 /label="cat promoter" /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS join(10977..11195,1..438) /label="CmR" /note="chloramphenicol acetyltransferase"