Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017310 | pEE491 | 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:
- pEE491
- Antibiotic Resistance:
- Kanamycin
- Length:
- 11610 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Plants
- Copy Number:
- Low
- Promoter:
- CaMV35S(enhanced)
- Growth Temperature:
- 37℃
pEE491 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.
pEE491 vector Sequence
LOCUS V017310 11610 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017310
VERSION V017310
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11610)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11610
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..770
/label="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 1330..1941
/gene="CP"
/label="Capsid protein"
/note="Capsid protein from Tobacco rattle virus (isolate
PpK20). Accession#: Q88897"
misc_RNA 2327..2402
/label="gRNA scaffold"
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 3540..3557
/label="6xHis"
/note="6xHis affinity tag"
terminator 3592..3844
/label="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 3866..3890
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 5191..5817
/label="pVS1 StaA"
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 6249..7319
/label="pVS1 RepA"
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 7388..7582
/label="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7926..8066
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(8252..8840)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10268..11059)
/label="KanR"
/note="aminoglycoside phosphotransferase"
misc_feature 11484..11508
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"