pAdTrack-CMV vector (V001435)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001435 pAdTrack-CMV 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:
pAdTrack-CMV
Antibiotic Resistance:
Kanamycin
Length:
9237 bp
Type:
Mammalian Expression, Adenoviral
Replication origin:
ori
Copy Number:
Low Copy
Cloning Method:
Restriction Enzyme

pAdTrack-CMV vector Map

pAdTrack-CMV9237 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200SV40 poly(A) signalCMV enhancerCMV promoterEGFPSV40 poly(A) signalAd5 PsiITRM13 oriNeoR/KanRoriL4440ITREarly 4 ORF1 proteinPackaging protein 1Hexon-interlacing proteinCMV enhancerCMV promoter

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.

pAdTrack-CMV vector Sequence

LOCUS       V001435                 9237 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001435
VERSION     V001435
KEYWORDS    pAdTrack-CMV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9237)
  AUTHORS   He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B
  TITLE     A simplified system for generating recombinant adenoviruses.
  JOURNAL   Proc Natl Acad Sci U S A. 1998 Mar 3. 95(5):2509-14.
   PUBMED   9482916
REFERENCE   2  (bases 1 to 9237)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9237)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
            Acad Sci U S A. 1998 Mar 3. 95(5):2509-14."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    5..126
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     enhancer        212..515
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        516..719
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             764..1480
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    1619..1740
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_signal     complement(1883..2033)
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     repeat_region   2121..2223
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     rep_origin      2261..2641
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2692..3483)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4315..4903
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     5057..5074
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     repeat_region   5163..5265
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     CDS             5575..5958
                     /note="Early 4 ORF1 protein from Human adenovirus C
                     serotype 2. Accession#: P03242"
                     /label="Early 4 ORF1 protein"
     CDS             6543..7877
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     CDS             complement(7943..8362)
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     enhancer        8524..8827
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8829..9028
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"