pGBW-m4092694 vector (V016844)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016844 pGBW-m4092694 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:
pGBW-m4092694
Antibiotic Resistance:
Ampicillin
Length:
8300 bp
Type:
Protein expression
Replication origin:
ori
Host:
E. coli
Promoter:
T7
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pGBW-m4092694 vector Map

pGBW-m40926948300 bp400800120016002000240028003200360040004400480052005600600064006800720076008000Transposase InsC for insertion element IS2T7 terminatortonB terminatorcat promoterAmpRropT7Te terminatororiT3Te terminator

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.

pGBW-m4092694 vector Sequence

LOCUS       V016844                 8300 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016844
VERSION     V016844
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8300)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3756..4118
                     /gene="insC1"
                     /label="Transposase InsC for insertion element IS2"
                     /note="Transposase InsC for insertion element IS2 from
                     Shigella flexneri. Accession#: P59444"
     terminator      5183..5230
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     terminator      5273..5304
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     promoter        5305..5407
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             5408..6265
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             6269..6457
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     terminator      6484..6511
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(6523..7110)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      complement(7132..7161)
                     /label="T3Te terminator"
                     /note="phage T3 early transcription terminator"