Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002331 | pUG6 | 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
Plasmid pUG6 carrying the loxP–kanMX–loxP module. LoxP-flanked marker gene deletion cassette: loxP-pAgTEF1-kanMX-tAgTEF1-loxP, selectable phenotype: G418 resistance
- Vector Name:
- pUG6
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4009 bp
- Type:
- PCR template vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Guldener U, Heck S, Fielder T, Beinhauer J, Hegemann JH.
- Promoter:
- TEF
pUG6 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
- Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH. A second set of loxP marker cassettes for Cre-mediated multiple gene knockouts in budding yeast. Nucleic Acids Res. 2002 Mar 15;30(6):e23.
pUG6 vector Sequence
LOCUS 40924_45503 4009 bp DNA circular SYN 18-DEC-2018
DEFINITION PCR template vector pUG6, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4009)
AUTHORS Guldener U, Heck S, Fielder T, Beinhauer J, Hegemann JH.
TITLE A new efficient gene disruption cassette for repeated use in budding
yeast
JOURNAL Nucleic Acids Res. 24 (13), 2519-2524 (1996)
PUBMED 8692690
REFERENCE 2 (bases 1 to 4009)
AUTHORS Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
TITLE A second set of loxP marker cassettes for Cre-mediated multiple gene
knockouts in budding yeast
JOURNAL Nucleic Acids Res. 30 (6), E23 (2002)
PUBMED 11884642
REFERENCE 3 (bases 1 to 4009)
AUTHORS Gueldener U, Hegemann JH, Heck S, Fiedler T, Beinhauer JD.
TITLE Direct Submission
JOURNAL Submitted (23-AUG-2000) Institut fuer Mikrobiologie,
Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
40225, Germany
REFERENCE 4 (bases 1 to 4009)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 4009)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "1996"; volume: "24"; issue: "13"; pages:
"2519-2524"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(23-AUG-2000) Institut fuer Mikrobiologie,
Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
40225, Germany"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4009
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 53..86
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
gene 141..1497
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
misc_feature 1560..1593
/label=loxP site
/note="loxP site"
protein_bind complement(1560..1593)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter complement(1647..1665)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(1923..2511)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2685..3542)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3543..3647)
/label=AmpR promoter
promoter 3993..4009
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"