pShuttle-CMV-GFP U6promoter vector (V017063)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017063 pShuttle-CMV-GFP U6promoter 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:
pShuttle-CMV-GFP U6promoter
Antibiotic Resistance:
Kanamycin
Length:
8233 bp
Type:
RNA interference
Replication origin:
ori
Host:
Mammalian cells, Adenovirus
Promoter:
U6
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pShuttle-CMV-GFP U6promoter vector Map

pShuttle-CMV-GFP U6promoter8233 bp400800120016002000240028003200360040004400480052005600600064006800720076008000U6 promoterCMV enhancerCMV promoterEGFPHexon-interlacing proteinPackaging protein 1Early 4 ORF1 proteinITRoriNeoR/KanRM13 oriITRAd5 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.

pShuttle-CMV-GFP U6promoter vector Sequence

LOCUS       V017063                 8233 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017063
VERSION     V017063
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8233)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..241
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        363..666
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        667..870
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             915..1631
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             1730..2149
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(2215..3549)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     CDS             complement(4134..4517)
                     /note="Early 4 ORF1 protein from Human adenovirus C
                     serotype 2. Accession#: P03242"
                     /label="Early 4 ORF1 protein"
     repeat_region   4827..4929
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     rep_origin      complement(5189..5777)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             6609..7400
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(7451..7831)
                     /direction=LEFT
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     repeat_region   7869..7971
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     misc_signal     8059..8209
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"