Restriction digests of the clone give the following sizes (kb): KpnI--6.3; KpnI/SstI--4.3, 2.0; ClaI--6.3. 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 LYS2 markers by a homologous recombination event). Deleter vector for constructing designer yeast strains with a non-revertable deletion of a lys2 auxotrophic marker. |
