Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010039 | CIp10 | 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
The CIp10 is an efficient and convenient integrating vector for Candida albicans.
- Vector Name:
- CIp10
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5198 bp
- Type:
- Integrating vector
- Replication origin:
- ori
- Source/Author:
- Murad AM, Lee PR, Broadbent ID, Barelle CJ, Brown AJ.
- Selection Marker:
- URA3
- Promoter:
- T3
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
CIp10 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.
References
- Murad AM, Lee PR, Broadbent ID, Barelle CJ, Brown AJ. CIp10, an efficient and convenient integrating vector for Candida albicans. Yeast. 2000;16(4):325-327. doi:10.1002/1097-0061(20000315)16:4<325::AID-YEA538>3.0.CO;2-#
CIp10 vector Sequence
LOCUS V010039 5198 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V010039
VERSION V010039
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 5198)
AUTHORS Murad AM, Lee PR, Broadbent ID, Barelle CJ, Brown AJ.
TITLE CIp10, an efficient and convenient integrating vector for Candida
albicans
JOURNAL Yeast 16 (4), 325-327 (2000)
PUBMED 10669870
REFERENCE 2 (bases 1 to 5198)
AUTHORS Abdul Murad AM, Lee PR, Broadbent ID, Brown AJP.
TITLE Direct Submission
JOURNAL Submitted (31-AUG-1999) Molecular and Cell Biology, University of
Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25
2ZD, UK
REFERENCE 3 (bases 1 to 5198)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5198)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast";
date: "2000"; volume: "16"; issue: "4"; pages: "325-327"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(31-AUG-1999) Molecular and Cell Biology, University of Aberdeen,
Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5198
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1124..1933
/gene="URA3"
/label="Orotidine 5'-phosphate decarboxylase"
/note="Orotidine 5'-phosphate decarboxylase from Candida
albicans (strain SC5314 / ATCC MYA-2876). Accession#:
P13649"
primer_bind 2060..2076
/label="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2585..2590
/label="StuI site for targeted integration"
/note="StuI site for targeted integration"
misc_feature 2602..2607
/label="NcoI site for targeted integration"
/note="NcoI site for targeted integration"
primer_bind complement(2964..2980)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3010..3028)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3049..3065)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3073..3089)
/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(3097..3127)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(3142..3163)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3451..4039)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4213..5070)
/label="AmpR"
/note="beta-lactamase"
promoter complement(5071..5175)
/label="AmpR promoter"