Basic Vector Information
- Vector Name:
- pOAD
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8429 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Uetz P, Giot L, Cagney G, Mansfield TA, Judson RS, Knight JR,
- Copy Number:
- High copy number
- Promoter:
- ADH1(long)
pOAD 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 ℃.
pOAD vector Sequence
LOCUS 40924_33707 8429 bp DNA circular SYN 01-JAN-1980
DEFINITION Yeast two-hybrid """"prey"""" vector for fusing a gene to the GAL4
activation domain.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8429)
AUTHORS Uetz P, Giot L, Cagney G, Mansfield TA, Judson RS, Knight JR,
Lockshon D, Narayan V, Srinivasan M, Pochart P, Qureshi-Emili A, Li
Y, Godwin B, Conover D, Kalbfleisch T, Vijayadamodar G, Yang M,
Johnston M, Fields S, Rothberg JM.
TITLE A comprehensive analysis of protein-protein interactions in
Saccharomyces cerevisiae.
JOURNAL Nature 2000;403:623-7.
PUBMED 10688190
REFERENCE 2 (bases 1 to 8429)
AUTHORS Fields Lab
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8429)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature";
date: "2000"; volume: "403"; pages: "623-7"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8429
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 408..595
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
misc_feature complement(1427..2589)
/label=CEN/ARS
/note="S. cerevisiae CEN4 centromere fused to the
autonomously replicating sequence ARS1/ARS416"
CDS complement(2954..4045)
/codon_start=1
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(4058..4462)
/label=LEU2 promoter
promoter 4568..4672
/label=AmpR promoter
CDS 4673..5530
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5704..6292
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 7281..7981
/label=ADH1 promoter
/note="promoter for alcohol dehydrogenase 1"
CDS 8026..8046
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 8062..8403
/codon_start=1
/label=GAL4 activation domain
/note="activation domain of the GAL4 transcriptional
activator"
/translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE"
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