pCAG-VSVL vector (V013826)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013826 pCAG-VSVL In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pCAG-VSVL
Antibiotic Resistance:
Ampicillin
Length:
11129 bp
Type:
Packaging assistance
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Promoter:
CAG
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pCAG-VSVL vector Map

pCAG-VSVL11129 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000chimeric intronRNA-directed RNA polymerase Lbeta-globin poly(A) signallac promoterCAP binding siteSV40 promoterSV40 poly(A) signaloriAmpRAmpR 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.

pCAG-VSVL vector Sequence

LOCUS       V013826                11129 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013826
VERSION     V013826
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11129)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          663..1680
                     /label="chimeric intron"
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             1740..8066
                     /gene="L"
                     /label="RNA-directed RNA polymerase L"
                     /note="RNA-directed RNA polymerase L from Vesicular
                     stomatitis Indiana virus (strain Mudd-Summers). Accession#:
                     Q98776"
     polyA_signal    8217..8272
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     promoter        complement(8681..8711)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8725..8746)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8804..9000
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     polyA_signal    9006..9140
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(9378..9966)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10140..10997)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(10998..11102)
                     /label="AmpR promoter"