Restriction digests of the clone give the following sizes (kb): EcoRI--8.5; HindIII--8.5; BamHI--8.5; SalI (uncut). 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. YCF4 is used to recover sequences distal to the mapping sequence. Shuttle chromosome fragmentation vector. Constructed by inserting a 1.4 kb fragment containing CEN4 at the SalI site (blunt-ended) of YCF3. The order of the major features in this plasmid is: Y' - MCS - CEN4 - URA3 - SUP11 - rep - pMB1 ori - bla. |
