rAAV2-retro helper vector (V000739)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V000739	rAAV2-retro helper	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:: rAAV2-retro helper

Antibiotic Resistance:: Ampicillin

Length:: 8369 bp

Type:: AAV, Unspecified ; helper vector

Replication origin:: ori

rAAV2-retro helper vector Map

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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.

rAAV2-retro helper vector Sequence

LOCUS       V000739                 8369 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000739
VERSION     V000739
KEYWORDS    rAAV2-retro helper
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8369)
  AUTHORS   Tervo DG, Hwang BY, Viswanathan S, Gaj T, Lavzin M, Ritola KD, Lindo
            S, Michael S, Kuleshova E, Ojala D, Huang CC, Gerfen CR, Schiller J,
            Dudman JT, Hantman AW, Looger LL, Schaffer DV, Karpova AY
  TITLE     A Designer AAV Variant Permits Efficient Retrograde Access to
            Projection Neurons.
  JOURNAL   Neuron. 2016 Oct 6. pii: S0896-6273(16)30580-3. doi:
            10.1016/j.neuron.2016.09.021.
   PUBMED   27720486
REFERENCE   2  (bases 1 to 8369)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8369)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron.
            2016 Oct 6. pii: S0896-6273(16)30580-3. doi:
            10.1016/j.neuron.2016.09.021."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   44..146
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     CDS             294..2150
                     /gene="Rep78"
                     /label="Protein Rep78"
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"
     CDS             4249..4260
                     /label="WELQut site"
                     /note="WELQut protease recognition and cleavage site"
     rep_origin      5186..5699
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6181..6200)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     6300..6322
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6360..6378)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        6446..6550
                     /label="AmpR promoter"
     CDS             6551..7408
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7582..8170
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     8324..8341
                     /label="L4440"
                     /note="L4440 vector, forward primer"