Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017025 | pN094 | 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:
- pN094
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6659 bp
- Type:
- Protein expression
- Replication origin:
- oriV
- Host:
- Plants
- Promoter:
- CaMV 35S
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pN094 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.
pN094 vector Sequence
LOCUS V017025 6659 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017025
VERSION V017025
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6659)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6659
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(76..100)
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
rep_origin 184..893
/label="oriV"
/note="incP origin of replication"
promoter 910..1014
/label="AmpR promoter"
CDS 1015..1872
/label="AmpR"
/note="beta-lactamase"
rep_origin 2046..2634
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 2970..4115
/label="trfA"
/note="trans-acting replication protein that binds to and
activates oriV"
misc_feature complement(4274..4298)
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
terminator complement(4347..5054)
/label="OCS terminator"
/note="octopine synthase terminator"
CDS complement(5082..6113)
/gene="npgA"
/label="4'-phosphopantetheinyl transferase NpgA"
/note="4'-phosphopantetheinyl transferase NpgA from
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS
112.46 / NRRL 194 / M139). Accession#: G5EB87"
misc_feature complement(6122..6175)
/label="TMV Omega"
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
promoter complement(6204..6549)
/label="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"