Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017406 | pCpf1-sp | 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:
- pCpf1-sp
- Antibiotic Resistance:
- Streptomycin
- Length:
- 12595 bp
- Type:
- Genome editing
- Replication origin:
- RSF1010 oriV
- Host:
- E. coli
- Copy Number:
- Low
- Promoter:
- J23119
- 5' Primer:
- M13F
- Growth Temperature:
- 37℃
pCpf1-sp 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.
pCpf1-sp vector Sequence
LOCUS V017406 12595 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017406
VERSION V017406
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12595)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12595
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 14..3913
/gene="cas12a"
/label="CRISPR-associated endonuclease Cas12a"
/note="CRISPR-associated endonuclease Cas12a from
Francisella tularensis subsp. novicida (strain U112).
Accession#: A0Q7Q2"
promoter 3923..3957
/label="J23119 promoter"
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
promoter 4212..4230
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(4278..4296)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter complement(4365..4395)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(4410..4431)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 5141..5929
/label="SmR"
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
rep_origin complement(6711..7105)
/direction=LEFT
/label="RSF1010 oriV"
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
oriT 7447..7534
/label="RSF1010 oriT"
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 8773..9741
/label="RSF1010 RepB"
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 10255..11091
/label="RSF1010 RepA"
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 11081..11929
/label="RSF1010 RepC"
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"