pCMV-APOPTIN(viru) vector (V014123)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014123 pCMV-APOPTIN(viru) 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:
pCMV-APOPTIN(viru)
Antibiotic Resistance:
Ampicillin
Length:
4367 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Promoter:
CMV
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pCMV-APOPTIN(viru) vector Map

pCMV-APOPTIN(viru)4367 bp600120018002400300036004200CMV enhancerCMV promoterchimeric intronT7 promoterApoptinSV40 poly(A) signalf1 oriAmpR promoterAmpRori

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.

pCMV-APOPTIN(viru) vector Sequence

LOCUS       V014123                 4367 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014123
VERSION     V014123
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4367)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          857..989
                     /label="chimeric intron"
                     /note="chimera between introns from human beta-globin and
                     immunoglobulin heavy chain genes"
     promoter        1034..1052
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1076..1438
                     /gene="VP3"
                     /label="Apoptin"
                     /note="Apoptin from Chicken anemia virus (isolate Germany
                     Cuxhaven-1). Accession#: Q99152"
     polyA_signal    complement(1481..1602)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1783..2238
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2570..2674
                     /label="AmpR promoter"
     CDS             2675..3532
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3706..4294
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"