Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006839 | pREP | 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:
- pREP
- Antibiotic Resistance:
- Kanamycin
- Length:
- 11214 bp
- Type:
- Plant Expression Vectors
- Replication origin:
- ori
- Host:
- Plants
- Promoter:
- CaMV 35S (enhanced)
pREP vector 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.
pREP vector Sequence
LOCUS V006839 11214 bp DNA circular SYN 13-JAN-2022
DEFINITION Exported.
ACCESSION V006839
VERSION V006839
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11214)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 11214)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11214
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(10..26)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(34..50)
/label="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(58..88)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(103..124)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 315..992
/label="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 1060..2082
/label="HygR"
/note="aminoglycoside phosphotransferase from E. coli"
polyA_signal 2125..2299
/label="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature complement(2377..2401)
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
CDS 2826..3617
/label="KanR"
/note="aminoglycoside phosphotransferase"
rep_origin 3707..4295
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(4481..4621)
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4965..5159)
/direction=LEFT
/label="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(5228..6292)
/label="pVS1 RepA"
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(6729..7355)
/label="pVS1 StaA"
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
misc_feature complement(8655..8679)
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 8882..8898
/label="M13 fwd"
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 9324..9668
/label="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 9728..9752
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 9761..10636
/note="Replication-associated protein A from Bean yellow
dwarf virus. Accession#: O39521"
/label="Replication-associated protein A"
protein_bind complement(10867..10891)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
terminator 10955..11207
/label="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"