Cyto-roGFP vector (V013211)

Price Information

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V013211 Cyto-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:
Cyto-roGFP
Antibiotic Resistance:
Ampicillin
Length:
6935 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Adenovirus
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

Cyto-roGFP vector Vector Map

Cyto-roGFP6935 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900oriAmpRAmpR promoterITRAd5 PsiCMV enhancerCMV promoterEGFPSV40 poly(A) signalHexon-interlacing proteinPackaging protein 1

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.

Cyto-roGFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V013211                 6935 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013211
VERSION     V013211
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6935)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(101..689)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(863..1720)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1721..1825)
                     /label="AmpR promoter"
     repeat_region   1939..2041
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     misc_signal     2129..2279
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        2308..2611
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2612..2815
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2870..3586
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    complement(3610..3744)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             4312..4731
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(4797..6131)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"