pAD4 (ATCC® 87470)

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

Permits and Restrictions

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Designations pAD4
Depositors JD Boeke
Biosafety Level 1
Host
Distribution host: Escherichia coli unknown (ATCC 33694)
Vector Information
Size (kb): 7.375
DESCRIPTION OF VECTOR:
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
Marker(s):URA3,ampR
Construct size (kb): 7.375
Features: marker(s): URA3
marker(s): ampR
other: MET15 deleter cassette
replicon: f1
replicon: pMB1
Applications
YI-type (integrating) shuttle vector
marker deletion vector O-acetylhomoserine sulfhydrylase
Comments
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
References

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

Related Products
component of:ATCC 87472
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Shipping Information Distributed: freeze-dried
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