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Distribution host: Escherichia coli unknown (ATCC 33694)

Size (kb): 6.485

DESCRIPTION OF VECTOR:
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

Marker(s):URA3,ampR

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.

Temperature: 37°C

component of:ATCC 87472