Restriction digests of the clone give the following sizes (kb): EcoRI/XhoI--7.3; KpnI--5.6, 1.5; BamHI/XbaI--6.4, 0.9. S. cerevisiae is cotransformed with the lambda clone of interest and vector linearized with an EcoRI/XhoI double digest. Ura+ colonies are tested for complementation of a mutant phenotype. YC-type shuttle vector designed to clone yeast genes that are difficult to clone by rescuing inserts from the vector lambdaMG3 by homologous recombination. Constructed by insertion of a 0.858 kb BamHI-XbaI fragment and a 1.5 kb KpnI fragment from lambdaMG3 into the corresponding sites of pRS316 (ATCC 77145). The order of the major features in this plasmid is: f1 ori - lacZ - T7 promoter - KpnI/lambdaMG3/KpnI - MCS - BamHI/lambdaMG3/XbaI - T3 promoter - pMB1 ori - ampR - CEN6/ARS4 - URA3. |
