Basic Vector Information
- Vector Name:
- pUG66
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3580 bp
- Type:
- PCR template vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
- Promoter:
- TEF
pUG66 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 ℃.
pUG66 vector Sequence
LOCUS 40924_45508 3580 bp DNA circular SYN 18-DEC-2018
DEFINITION PCR template vector pUG66, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3580)
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 2 (bases 1 to 3580)
AUTHORS Gueldener U, Heinisch JH, Voss D, Koehler G, Hegemann JH.
TITLE Direct Submission
JOURNAL Submitted (23-AUG-2000) Institut fuer Mikrobiologie,
Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
40225, Germany
REFERENCE 3 (bases 1 to 3580)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3580)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(23-AUG-2000) Institut fuer Mikrobiologie,
Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
40225, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3580
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 54..87
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 142..485
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 486..866
/codon_start=1
/gene="ble"
/product="bleomycin resistance protein"
/label=ble
/protein_id="AAG34545.1"
/translation="MADQATPNLPSRDFDSTAAFYERLGFGIVFRDAGWMILQRGDLKL
EFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGTMA
ALVDPDGTLLRLIQNELLAGIS"
gene 486..866
/gene="ble"
/label=ble
terminator 872..1069
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_feature 1132..1165
/label=loxP site
/note="loxP site"
protein_bind complement(1132..1165)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter complement(1219..1237)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(1495..2083)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2257..3114)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3115..3219)
/label=AmpR promoter
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