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pAAV2/2 vector (V001085)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001085 pAAV2/2 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:
pAAV2/2
Antibiotic Resistance:
Ampicillin
Length:
7468 bp
Type:
Mammalian Expression, AAV
Replication origin:
ori
Copy Number:
High Copy
Promoter:
p5
5' Primer:
M13 Reverse

pAAV2/2 vector Map

Copy Sequence View Map
pAAV2/27468 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200pBR322ori-FL4440CAP binding sitelac promoterlac operatorM13 revSK primerFRTProtein Rep78Capsid protein VP1M13 fwdf1 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.

pAAV2/2 vector Sequence

LOCUS       V001085                 7468 bp    DNA     circular SYN 19-DEC-2023
DEFINITION  Exported.
ACCESSION   V001085
VERSION     V001085
KEYWORDS    pAAV2-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7468)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7468)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label="pBR322ori-F"
                     /note="pBR322ori-F"
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label="L4440"
                     /note="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        555..585
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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)."
     primer_bind     617..633
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     683..699
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    713..760
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             919..2781
                     /gene="Rep78"
                     /label="Protein Rep78"
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"
     CDS             2801..5005
                     /gene="VP1"
                     /label="Capsid protein VP1"
                     /note="Capsid protein VP1 from Adeno-associated virus 2
                     (isolate Srivastava/1982). Accession#: P03135"
     primer_bind     complement(5336..5352)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      complement(5493..5948)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5975..6079
                     /label="AmpR promoter"
     CDS             6080..6937
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7111..7468
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"