pAD1 (ATCC® 87468)

Applications: YI-type (integrating) shuttle vectormarker deletion vector 3-isopropylmalate dehydrogenase beta-isopropylmalate dehydrogenase  /  Depositors: JD Boeke

Designations pAD1
Permits and Restrictions

View Permits

Depositors JD Boeke
Biosafety Level 1
Distribution host: Escherichia coli unknown (ATCC 33694)
Vector Information
Size (kb): 6.485
Intact vector size: 6.485
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: LEU2 deleter cassette
replicon: pMB1
marker(s): ampR, <-
Vector: pAD1 (phagemid)
Construction: pRS406
Construct size (kb): 6.485
Features: marker(s): URA3
marker(s): ampR
other: LEU2 deleter cassette
replicon: f1
replicon: pMB1
YI-type (integrating) shuttle vector
marker deletion vector 3-isopropylmalate dehydrogenase beta-isopropylmalate dehydrogenase
Restriction digests of the clone give the following sizes (kb): KpnI--6.5; KpnI/SstI--4.3, 2.2; ClaI--6.5.
The two step selection process requires a ura3 transformation host (this host can be created using pJL164 (ATCC 87471)). After transformation with the SalI 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 LEU2 markers by a homologous recombination event).
This deleter vector is used to create designer yeast strains with a non-revertable leu2 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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