pMV306hsp vector (V017276) Gene synthesis in pMV306hsp backbone

Price Information

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

It is a bacterial expression plasmid. It contains the hsp60 promoter from Mycobacterium bovis BCG, which can drive the expression of inserted genes. It's often used to construct recombinant plasmids with specific genes so that the target genes can be expressed in host cells.

pMV306hsp vector Map

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.

References

• Andreu et al PLoS One. 2010 May 24;5(5):e10777.

pMV306hsp vector Sequence

LOCUS       Exported                4356 bp DNA     circular SYN 13-JAN-2025
DEFINITION  Exported.
ACCESSION   V017276
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4356)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4356)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(2451..4356,1..2450)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..813
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1148..1736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      2382..2428
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             2961..4073
                     /gene="33"
                     /label=Integrase
                     /note="Integrase from Mycobacterium phage L5. Accession#: 
                     P22884"