Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V000820 | pCG | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pCG
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5015 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Promoter:
- SV40
pCG 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.
pCG vector Sequence
LOCUS V000820 5015 bp DNA circular SYN 13-JAN-2022 DEFINITION Exported. ACCESSION V000820 VERSION V000820 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 5015) TITLE Direct Submission REFERENCE 2 (bases 1 to 5015) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5015 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 304..507 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" 5'UTR 597..636 /label="HSV TK 5'-UTR" /note="5' untranslated region from the herpes simplex virus thymidine kinase gene" intron 704..1276 /label="beta-globin intron" /note="intron from rabbit beta-globin gene" CDS 1280..1402 /gene="HBB" /label="Hemoglobin subunit beta" /note="Hemoglobin subunit beta from Colobus guereza. Accession#: Q9TT33" polyA_signal 1473..1528 /label="beta-globin poly(A) signal" /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" promoter complement(1865..2061) /label="SV40 promoter" /note="SV40 early promoter" rep_origin complement(2299..2887) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3061..3918) /label="AmpR" /note="beta-lactamase" promoter complement(3919..4023) /label="AmpR promoter" rep_origin complement(4355..4810) /direction=LEFT /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 4955..4971 /label="M13 fwd" /note="common sequencing primer, one of multiple similar variants" promoter 4978..4996 /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" enhancer 5015 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer"