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. |
