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pAAV2/7 vector (V012817)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012817 pAAV2/7 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

AAV packaging plasmid, expressing Rep2 and Cap7

Vector Name:
pAAV2/7
Antibiotic Resistance:
Ampicillin
Length:
7333 bp
Type:
Viral Expression & Packaging Vectors
Replication origin:
ori
Promoter:
Truncated P5
Growth Strain(s):
DH5alpha
Growth Temperature:
37℃

pAAV2/7 vector Map

Copy Sequence View Map
pAAV2/77333 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200M13 fwdT7 promoterKS primerProtein Rep78WELQut siteT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterf1 ori

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.

pAAV2/7 vector Sequence

LOCUS       V012817                 7333 bp    DNA     circular SYN 21-AUG-2023
DEFINITION  Exported.
ACCESSION   V012817
VERSION     V012817
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7333)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..28
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        38..56
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     86..102
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             157..2019
                     /gene="Rep78"
                     /label="Protein Rep78"
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"
     CDS             4094..4105
                     /label="WELQut site"
                     /note="WELQut protease recognition and cleavage site"
     promoter        complement(4557..4575)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4596..4612)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4620..4636)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4644..4674)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4689..4710)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4998..5586)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5760..6617)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6618..6722)
                     /label="AmpR promoter"
     rep_origin      complement(6748..7203)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"