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Matrix-roGFP vector (V001383)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001383 Matrix-roGFP 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:
Matrix-roGFP
Antibiotic Resistance:
Ampicillin
Length:
6929 bp
Type:
Mammalian Expression, Adenoviral
Replication origin:
ori
Copy Number:
High Copy
Promoter:
CMV
Cloning Method:
Restriction Enzyme
5' Primer:
5'GGCGTGTACGGTGGGAGGTCTATA-3'
3' Primer:
5'-TTAAAAAACCTCCCCACCTCCCCC-3'

Matrix-roGFP vector Map

Copy Sequence View Map
Matrix-roGFP6929 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900CMV enhancerCMV promoterEGFPSV40 poly(A) signalpGEX 3'Hexon-interlacing proteinPackaging protein 1oriAmpRAmpR promoterpBRforEcoITRAd5 Psi

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.

Matrix-roGFP vector Sequence

LOCUS       V001383                 6929 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001383
VERSION     V001383
KEYWORDS    Matrix-roGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6929)
  AUTHORS   Waypa GB, Marks JD, Guzy R, Mungai PT, Schriewer J, Dokic D,
            Schumacker PT
  TITLE     Hypoxia triggers subcellular compartmental redox signaling in
            vascular smooth muscle cells.
  JOURNAL   Circ Res. 2010 Feb 19;106(3):526-35. doi:
            10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17.
   PUBMED   20019331
REFERENCE   2  (bases 1 to 6929)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6929)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1161/CIRCRESAHA.109.206334"; journalName: "Circ Res"; date:
            "2010-02-19- 19"; volume: "106"; issue: "3"; pages: "526-35"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..315
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        316..519
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             600..1316
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    complement(1360..1494)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1637..1659)
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     CDS             2062..2481
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(2547..3881)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     rep_origin      complement(4734..5322)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5496..6353)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6354..6458)
                     /label="AmpR promoter"
     primer_bind     6526..6544
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     repeat_region   6572..6674
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     misc_signal     6762..6912
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"