Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V015036 | pBE2R-mRFP1 | 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:
- pBE2R-mRFP1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8215 bp
- Type:
- Protein expression, Secretory expression
- Replication origin:
- ori
- Host:
- Bacillus subtilis
- Promoter:
- SP6
- Growth Temperature:
- 37℃
pBE2R-mRFP1 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.
pBE2R-mRFP1 vector Sequence
LOCUS V015036 8215 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V015036 VERSION V015036 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 8215) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..8215 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 891..1479 /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3531..4178) /gene="cat" /label="Chloramphenicol acetyltransferase" /note="Chloramphenicol acetyltransferase from Staphylococcus aureus. Accession#: P00485" CDS complement(4504..5505) /label="repB" /note="RepB replication protein" CDS 6273..6947 /label="mRFP1" /note="monomeric derivative of DsRed (Campbell et al., 2002)" terminator 6995..7066 /label="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 7082..7109 /label="T7Te terminator" /note="phage T7 early transcription terminator" promoter complement(7167..7185) /label="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" promoter 7970..8074 /label="AmpR promoter" CDS join(8075..8215,1..717) /label="AmpR" /note="beta-lactamase"