pAL119-TK vector (V008719)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V008719	pAL119-TK	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:: pAL119-TK

Antibiotic Resistance:: Ampicillin

Length:: 8478 bp

Type:: Adenoviral

Replication origin:: ori

Copy Number:: Low Copy

Cloning Method:: Restriction Enzyme

pAL119-TK vector Map

Copy Sequence View Map

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.

pAL119-TK vector Sequence

LOCUS       V008719                 8478 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V008719
VERSION     V008719
KEYWORDS    pAL119-TK
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8478)
  AUTHORS   Dewey RA, Morrissey G, Cowsill CM, Stone D, Bolognani F, Dodd NJ,
            Southgate TD, Klatzmann D, Lassmann H, Castro MG, Loewenstein PR
  TITLE     Chronic brain inflammation and persistent herpes simplex virus 1
            thymidine kinase expression in survivors of syngeneic glioma treated
            by adenovirus-mediated gene therapy: implications for clinical
            trials.
  JOURNAL   Nat Med. 1999 Nov . 5(11):1256-63.
   PUBMED   10545991
REFERENCE   2  (bases 1 to 8478)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8478)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Med.
            1999 Nov . 5(11):1256-63."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..103
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     misc_signal     191..341
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     promoter        577..780
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     777..801
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     CDS             876..2003
                     /label="HSV TK"
                     /note="herpes simplex virus thymidine kinase"
     CDS             2587..3006
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(3072..4406)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     CDS             6030..6218
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     primer_bind     complement(6215..6237)
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     misc_feature    6323..6463
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6478..6495)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6649..7237)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7411..8268)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8269..8373)
                     /label="AmpR promoter"
     primer_bind     8441..8459
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"