Basic Vector Information
pUG74 vector Vector Map
Plasmid Resuspension 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 ℃.
pUG74 vector Sequence
LOCUS 40924_45513 3772 bp DNA circular SYN 18-DEC-2018
DEFINITION PCR template vector pUG74, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3772)
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 3772)
AUTHORS Hegemann JH, Heick SB.
TITLE Delete and Repeat: A Comprehensive Toolkit for Sequential Gene
Knockout in the Budding Yeast Saccharomyces cerevisiae
JOURNAL Methods Mol. Biol. 765, 189-206 (2011)
PUBMED 21815094
REFERENCE 3 (bases 1 to 3772)
AUTHORS Hegemann JH, Heick SB.
TITLE Direct Submission
JOURNAL Submitted (13-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung
der Mikroorganismen, Heinrich-Heine-Universitaet,
Universitatsstrasse 1, Duesseldorf, NRW 40225, Germany
REFERENCE 4 (bases 1 to 3772)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 3772)
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: "Methods
Mol. Biol. 765, 189-206 (2011)"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(13-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung der
Mikroorganismen, Heinrich-Heine-Universitaet, Universitatsstrasse 1,
Duesseldorf, NRW 40225, Germany"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3772
/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..1260
/label=natMX6
/note="yeast selectable marker conferring nourseothricin
resistance"
misc_feature 1323..1356
/label=loxP site
/note="loxP site"
protein_bind complement(1323..1356)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter complement(1410..1428)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(1686..2274)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2448..3305)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3306..3410)
/label=AmpR promoter
promoter 3756..3772
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
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