pAD4 (ATCC® 87470)

Applications: YI-type (integrating) shuttle vectormarker deletion vector O-acetylhomoserine sulfhydrylase  /  Depositors: JD Boeke

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Designations pAD4
Depositors JD Boeke
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Distribution host: Escherichia coli unknown (ATCC 33694)
Vector Information
Size (kb): 7.375
Intact vector size: 7.375
Type of vector: phagemid
Cloning sites:
Polylinker sites:
Other unique sites: BamHI
Construction: pRS406
Host range: Saccharomyces cerevisiae; Escherichia coli
Features (with orientation and position when available):
marker(s): URA3, ->
replicon: f1, <-
other: MET15 deleter cassette
replicon: pMB1
marker(s): ampR, <-
Vector: pAD4 (phagemid)
Construction: pRS406
Construct size (kb): 7.375
Features: marker(s): URA3
marker(s): ampR
other: MET15 deleter cassette
replicon: f1
replicon: pMB1
YI-type (integrating) shuttle vector
marker deletion vector O-acetylhomoserine sulfhydrylase
Restriction digests of the clone give the following sizes (kb): KpnI--4.2, 3.1; KpnI/SacI--4.2, 3.1, 0.03; ClaI--7.4.
The two step selection process requires a ura3 transformation host (this host can be created using pJL164 (ATCC 87471)). After transformation with the ClaI linearized vector, URA3 integrants are selected on ura- plates.
The designer deletion strain is then recovered by selection on 5-FOA plates (loss of URA3 and MET15 markers by a homologous recombination event).
MET15, MET17 and MET25 are synonymous.
This deleter vector is used to create designer yeast strains with a non-revertable met15 auxotrophic marker deletion.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37°C

Brachmann CB, et al. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14: 115-132, 1998. PubMed: 9483801

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component of:ATCC 87472
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