Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006311 | pFA6a-VC-His3MX6 | 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:
- pFA6a-VC-His3MX6
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4318 bp
- Type:
- Yeast tagging vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Sung MK, Huh WK.
- Promoter:
- TEF
pFA6a-VC-His3MX6 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.
pFA6a-VC-His3MX6 vector Sequence
LOCUS 40924_19631 4318 bp DNA circular SYN 18-DEC-2018
DEFINITION Yeast tagging vector pFA6a-VC-His3MX6, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4318)
AUTHORS Sung MK, Huh WK.
TITLE Bimolecular fluorescence complementation analysis system for in vivo
detection of protein-protein interaction in Saccharomyces cerevisiae
JOURNAL Yeast 24 (9), 767-775 (2007)
PUBMED 17534848
REFERENCE 2 (bases 1 to 4318)
AUTHORS Sung M-K., Huh W-K.
TITLE Direct Submission
JOURNAL Submitted (09-JAN-2007) School of Biological Sciences, Seoul
National University, Seoul 151-747, Republic of Korea
REFERENCE 3 (bases 1 to 4318)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4318)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast";
date: "2007"; volume: "24"; issue: "9"; pages: "767-775"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-JAN-2007) School of Biological Sciences, Seoul National
University, Seoul 151-747, Republic of Korea"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4318
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 114..365
/codon_start=1
/label=VC155
/note="C-terminal fragment of mVenus for use in bimolecular
fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
/translation="DKQKNGIKLNFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
terminator 389..576
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
gene 651..1851
/label=HIS3MX6
/note="yeast selectable marker encoding the S. pombe his5
gene, which corresponds to S. cerevisiae HIS3"
promoter complement(1956..1974)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(2232..2820)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2994..3851)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3852..3956)
/label=AmpR promoter
promoter 4302..4318
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"