Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010838 | pKANNIBAL | 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:
- pKANNIBAL
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6063 bp
- Type:
- Plant Vectors
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding
- Copy Number:
- High copy number
- Promoter:
- CaMV 35S
pKANNIBAL 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.
pKANNIBAL vector Sequence
LOCUS pKANNIBAL. 6063 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector for silencing genes in plants using intron-containing hairpin
RNA (hpRNA).
ACCESSION .
VERSION .
KEYWORDS pKANNIBAL
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6063)
AUTHORS Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding
PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP,
Green AG, Waterhouse PM.
TITLE Construct design for efficient, effective and high-throughput gene
silencing in plants.
JOURNAL Plant J. 2001;27:581-90.
PUBMED 11576441
REFERENCE 2 (bases 1 to 6063)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6063)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2001"; volume: "27"; pages: "581-90"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6063
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(19..35)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(215..643)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 670..774
/label=AmpR promoter
CDS 1075..1866
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 2045..2633
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2921..2942
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2957..2987
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2995..3011
/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)."
primer_bind 3019..3035
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 3066..3084
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 4102..4447
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 4449..4467
/label=MCS 1
/note="MCS 1"
/note="multiple cloning site 1"
intron 4475..5241
/label=PDK intron
/note="pyruvate orthophosphate dikinase intron from
Flaveria trinervia"
misc_feature 5261..5284
/label=MCS 2
/note="MCS 2"
/note="multiple cloning site 2"
terminator 5288..5994
/label=OCS terminator
/note="octopine synthase terminator"
promoter complement(6035..6053)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"