Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V016821 | P362-FBP_12 | 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:
- P362-FBP_12
- Antibiotic Resistance:
- Kanamycin
- Length:
- 22699 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Plants
- Selection Marker:
- Neo/G418
- Promoter:
- CaMV35S(enhanced)
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
P362-FBP_12 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.
P362-FBP_12 vector Sequence
LOCUS V016821 22699 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016821
VERSION V016821
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 22699)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..22699
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(5..21)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 224..248
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 1549..2175
/label="pVS1 StaA"
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 2607..3677
/label="pVS1 RepA"
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 3746..3940
/label="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 4284..4424
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4610..5198)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5288..6079)
/label="KanR"
/note="aminoglycoside phosphotransferase"
misc_feature 6504..6528
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(6606..6780)
/label="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6840..7628)
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
promoter complement(7697..8373)
/label="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 8564..8585
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8600..8630
/label="lac promoter"
/note="promoter for the E. coli lac operon"
CDS 9547..10413
/gene="cph"
/label="Caffeoylpyruvate hydrolase"
/note="Caffeoylpyruvate hydrolase from Neonothopanus nambi.
Accession#: A0A3G9JYJ6"
promoter 11105..11151
/label="minimal CaMV 35S promoter"
/note="minimal 35S promoter from cauliflower mosaic virus"
misc_feature 11182..11235
/label="TMV Omega"
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
CDS 11244..12275
/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"
terminator 12304..12504
/label="gene 7 terminator"
/note="transcription terminator for octopine-type Ti
plasmid gene 7"
promoter 12541..12921
/label="MAS promoter"
/note="mannopine synthase promoter (Velten et al., 1984)"
CDS 12923..14188
/gene="h3h"
/label="Hispidin-3-hydroxylase"
/note="Hispidin-3-hydroxylase from Neonothopanus nambi.
Accession#: A0A3G9K5C8"
terminator 14193..14445
/label="MAS terminator"
/note="mannopine synthase terminator"
promoter 14557..14902
/label="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
polyA_signal 20109..20283
/label="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
promoter 20804..21149
/label="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 21160..21960
/gene="luz"
/label="Luciferase"
/note="Luciferase from Neonothopanus nambi. Accession#:
A0A3G9JYH7"
terminator 21971..22678
/label="OCS terminator"
/note="octopine synthase terminator"