Shh10 vector (V016682)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016682 Shh10 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:
Shh10
Antibiotic Resistance:
Ampicillin
Length:
8342 bp
Type:
Packaging assistance
Replication origin:
ori
Host:
Mammalian cells, Adeno-associated virus
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

Shh10 vector Map

Shh108342 bp400800120016002000240028003200360040004400480052005600600064006800720076008000M13 oriWELQut siteProtein Rep78ITRoriAmpRAmpR 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.

Shh10 vector Sequence

LOCUS       V016682                 8342 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016682
VERSION     V016682
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8342)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(653..1166)
                     /direction=LEFT
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2092..2103)
                     /label="WELQut site"
                     /note="WELQut protease recognition and cleavage site"
     CDS             complement(4175..6031)
                     /gene="Rep78"
                     /label="Protein Rep78"
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"
     repeat_region   6179..6281
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     rep_origin      complement(6524..7112)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7286..8143)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8144..8248)
                     /label="AmpR promoter"