Basic Vector Information
pUG75 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 ℃.
pUG75 vector Sequence
LOCUS 40924_45518 4228 bp DNA circular SYN 18-DEC-2018 DEFINITION PCR template vector pUG75, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4228) 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 4228) 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 4228) 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 4228) TITLE Direct Submission REFERENCE 5 (bases 1 to 4228) 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..4228 /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..1716 /label=hphMX6 /note="yeast selectable marker conferring hygromycin resistance" misc_feature 1779..1812 /label=loxP site /note="loxP site" protein_bind complement(1779..1812) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter complement(1866..1884) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" rep_origin complement(2142..2730) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2904..3761) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3762..3866) /label=AmpR promoter promoter 4212..4228 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase"
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