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pAnc80L65 vector (V012623)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012623 pAnc80L65 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:
pAnc80L65
Antibiotic Resistance:
Kanamycin
Length:
7446 bp
Type:
Mammalian Expression, AAV, Synthetic Biology
Replication origin:
ori
Copy Number:
High Copy
Promoter:
Rep2
5' Primer:
M13rev

pAnc80L65 vector Map

Copy Sequence View Map
pAnc80L657446 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200WELQut siteM13 revoriKanRT7Te terminatorf1 oriM13 fwdKS primerProtein Rep78

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.

pAnc80L65 vector Sequence

LOCUS       V012623                 7446 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V012623
VERSION     V012623
KEYWORDS    pAnc80L65
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7446)
  AUTHORS   Zinn E, Pacouret S, Khaychuk V, Turunen HT, Carvalho LS,
            Andres-Mateos E, Shah S, Shelke R, Maurer AC, Plovie E, Xiao R,
            Vandenberghe LH
  TITLE     In Silico Reconstruction of the Viral Evolutionary Lineage Yields a
            Potent Gene Therapy Vector.
  JOURNAL   Cell Rep. 2015 Jul 28. pii: S2211-1247(15)00759-7. doi:
            10.1016/j.celrep.2015.07.019.
   PUBMED   26235624
REFERENCE   2  (bases 1 to 7446)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7446)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.
            2015 Jul 28. pii: S2211-1247(15)00759-7. doi:
            10.1016/j.celrep.2015.07.019."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2373..2384
                     /label="WELQut site"
                     /note="WELQut protease recognition and cleavage site"
     primer_bind     complement(2862..2878)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      complement(3194..3782)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4062..4874)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     terminator      complement(5091..5118)
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5154..5609)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5751..5767
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     5802..5818
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             join(5885..7446,1..301)
                     /gene="Rep78"
                     /label="Protein Rep78"
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"