Restriction digests of the clone give the following sizes (kb): EcoRI--7.1; HindIII--7.1; BamHI--7.1; HindIII/BglII--7.1. The sequence to be mapped is cloned into the MCS of both YCF3 (ATCC 77155) and YCF4 (ATCC 77156) (both orientations being produced). NotI is used to linearize the constructs, which are transformed independently into YPH49. YCF3 is used to recover sequences proximal to the mapping sequence. Shuttle chromosome fragmentation vector. Constructed by replacing the 346 bp HindIII/BamHI fragment from YRp14 with a 1.5 kb HindIII/BglII fragment containing the telomere-adjacent sequence (Y') and a polylinker. EcoRI and HindIII sites were removed. The order of the major features in this plasmid is: Y' - MCS - SalI - URA3 - SUP11 - rep - pMB1 ori - bla. |
