pAAV-RC vector (V009666)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V009666	pAAV-RC	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:: pAAV-RC

Antibiotic Resistance:: Ampicillin

Length:: 7327 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Grafsky AJ III.

Promoter:: lac

pAAV-RC 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.

pAAV-RC vector Sequence

LOCUS       V009666                 7327 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009666
VERSION     V009666
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7327)
  AUTHORS   Grafsky AJ III.
  TITLE     pAAV-RC for AAV Helper-Free System
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7327)
  AUTHORS   Grafsky AJ III.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2001) Technical Services, Stratagene, 11011 N.
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 7327)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7327)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (16-APR-2001) Technical Services, Stratagene, 11011 N. Torrey Pines
            Rd., La Jolla, CA 92037, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..52
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    66..113
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             131..1993
                     /gene="Rep78"
                     /label="Protein Rep78"
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"
     CDS             2013..4217
                     /gene="VP1"
                     /label="Capsid protein VP1"
                     /note="Capsid protein VP1 from Adeno-associated virus 2
                     (isolate Srivastava/1982). Accession#: P03135"
     primer_bind     complement(4548..4564)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      complement(4705..5160)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5187..5291
                     /label="AmpR promoter"
     CDS             5292..6149
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6323..6911
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    7199..7220
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7235..7265
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7273..7289
                     /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     7297..7313
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"