pCG vector (V000820)

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

pCG5015 bp6001200180024003000360042004800CMV enhancerCMV promoter5'UTRbeta-globin intronHemoglobin subunit betabeta-globin poly(A) signalSV40 promoteroriAmpRAmpR promoterf1 oriM13 fwdT7 promoter

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"