Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V003187 | pSilent-1 | 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:
- pSilent-1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7056 bp
- Type:
- Ascomycete silencing vector
- Replication origin:
- ori
- Source/Author:
- Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.
- Promoter:
- trpC
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pSilent-1 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.
pSilent-1 vector Sequence
LOCUS 40924_40392 7056 bp DNA circular SYN 18-DEC-2018
DEFINITION Ascomycete silencing vector pSilent-1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7056)
AUTHORS Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.
TITLE RNA silencing as a tool for exploring gene function in ascomycete
fungi
JOURNAL
REFERENCE 2 (bases 1 to 7056)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 7056)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (15-MAR-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 4 (bases 1 to 7056)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 7056)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Fungal
genetics and biology : FG "
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(15-MAR-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7056
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 24..128
/label=AmpR promoter
CDS 129..986
/label=AmpR
/note="beta-lactamase"
rep_origin 1160..1748
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/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 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2234..2588
/label=trpC promoter
/note="promoter for Aspergillus nidulans trpC"
CDS 2593..3615
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
regulatory 3638..4123
/label=trpC terminator
/note="trpC terminator"
/regulatory_class="terminator"
primer_bind 4132..4148
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5444..5460)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
intron 5491..5624
/note="CUT intron"
terminator 5829..6393
/label=trpC terminator
/note="transcription terminator from the Aspergillus
nidulans trpC gene"
promoter complement(6413..6431)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6441..6457)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6599..7054
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"