Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005901 | pgR106 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pgR106
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10451 bp
- Type:
- Transformation vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Lu R, Malcuit I, Moffett P, Ruiz MT, Peart J, Wu AJ, Rathjen JP, Bendahmane A, Day L, Baulcombe DC.
- Promoter:
- CaMV 35S
pgR106 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.
pgR106 vector Sequence
LOCUS V005901 10451 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V005901
VERSION V005901
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10451)
AUTHORS Lu R, Malcuit I, Moffett P, Ruiz MT, Peart J, Wu AJ, Rathjen JP,
Bendahmane A, Day L, Baulcombe DC.
TITLE High throughput virus-induced gene silencing implicates heat shock
protein 90 in plant disease resistance
JOURNAL EMBO J. 22 (21), 5690-5699 (2003)
PUBMED 14592968
REFERENCE 2 (bases 1 to 10451)
AUTHORS Lu R, Malcuit I, Moffett P, Ruiz T, Peart J, Wu A-J., Rathjen J,
Bendahmane A, Day L, Baulcombe D.
TITLE Direct Submission
JOURNAL Submitted (13-MAY-2003) Department of Plant Pathology, University of
California Riverside, 900 University Avenue, Riverside, CA 92521,
USA
REFERENCE 3 (bases 1 to 10451)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10451)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J.";
date: "2003"; volume: "22"; issue: "21"; pages: "5690-5699"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-MAY-2003) Department of Plant Pathology, University of
California Riverside, 900 University Avenue, Riverside, CA 92521,
USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10451
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 540..562
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
promoter 659..1002
/label="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 1087..5454
/note="RNA replication protein from Potato virus X (strain
X3). Accession#: P17779"
/label="RNA replication protein"
CDS 5488..6162
/note="Movement and silencing protein TGBp1 from Potato
virus X (strain X3). Accession#: P17780"
/label="Movement and silencing protein TGBp1"
misc_feature 6670..6702
/label="multiple cloning site"
/note="multiple cloning site"
misc_feature 6703..7654
/label="Potato virus X strain UK3 3'-sequence"
/note="Potato virus X strain UK3 3'-sequence"
terminator 7679..7931
/label="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 7935..8102
/label="shuttle vector pSK5640 sequence"
/note="shuttle vector pSK5640 sequence"
primer_bind complement(7938..7954)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(8121..8139)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8160..8176)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8184..8200
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(8208..8238)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(8253..8274)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 8513..8537
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
rep_origin complement(8628..9216)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9320..10132)
/label="KanR"
/note="aminoglycoside phosphotransferase"