Restriction digests of the clone give the following sizes (kb): BglII/BamHI--5.2, 3.8; BamHI--9.0. Restriction digests of the clone give the following sizes (kb): BglII--9.0; BamHI--9.0; BamHI/BglII--5.2, 3.8; SmaI--9.0. Used by inserting the excised cassette into a cloned target gene, isolating a restriction fragment containing the cassette with flanking target gene sequences, transforming Ura3- strains, and selecting for stable integrants. A plasmid containing an excisable URA3 cartridge for gene disruption that permits repeated use of URA3 selection in constructing multiply disrupted yeast strains. A derivative of pNKY51 with kanR from Tn903 inserted at the SmaI site of pNKY51. The order of the major features in this plasmid is: BglII (nt 1) - hisG - nt 1150 - URA3 - nt 2350 - kanR - nt 3850 - hisG - BamHI (nt 5000) - pMB1 ori - ampR. |
