Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002123 | pWSK29 | 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:
- pWSK29
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5434 bp
- Type:
- Cloning vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Wang RF, Kushner SR.
pWSK29 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.
pWSK29 vector Sequence
LOCUS 40924_46628 5434 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pWSK29, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5434)
AUTHORS Wang RF, Kushner SR.
TITLE Construction of versatile low-copy-number vectors for cloning,
sequencing and gene expression in Escherichia coli
JOURNAL Gene 100, 195-199 (1991)
PUBMED 2055470
REFERENCE 2 (bases 1 to 5434)
AUTHORS Maples VF, Kushner SR.
TITLE Direct Submission
JOURNAL Submitted (01-AUG-1997) Genetics, University of Georgia, Life
Sciences Bldg., Athens, GA 30602, USA
REFERENCE 3 (bases 1 to 5434)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5434)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene 100,
195-199 (1991)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-AUG-1997) Genetics, University of Georgia, Life Sciences Bldg.,
Athens, GA 30602, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5434
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(147..1004)
/label=AmpR
/note="beta-lactamase"
promoter complement(1005..1109)
/label=AmpR promoter
rep_origin complement(1135..1590)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 1732..1748
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1758..1776
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(1785..1892)
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(1905..1923)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1944..1960)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1968..1984)
/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)."
promoter complement(1992..2022)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2037..2058)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
misc_difference 2641
/label=pSC101
/note="pSC101"
misc_difference 2830
/replace="c"
/label=pSC101
/note="pSC101"
rep_origin 3337..3559
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 3607..4554
/label=Rep101
/note="RepA protein needed for replication with the pSC101
origin"
misc_difference 4633
/replace="g"
/label=pSC101
/note="pSC101"
misc_difference 4723
/replace="t"
/label=pSC101
/note="pSC101"
misc_difference 4997..4999
/label=pSC101
/note="pSC101"
misc_feature 5271..5431
/note="originates from pLG339; contains portion of tn903
(kanamycin resistance transposon)"