Restriction digests of the clone give the following sizes (kb): EcoRI--4.1, 2.8, 2.6, 1.1; HindIII--6.6, 2.8, 1.4; PstI--9.5, 1.0. pRS319a differs from pRS319 in that pRS319 has a truncated CAN1 gene and pRS319a has a full length CAN1 gene. A plasmid shuffling vector. YC-type centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains CAN1 marker (full length CAN1 gene) for negative selection. pRS319a was constructed by inserting a 4.161 kb PvuII-Sma CAN1 fragment from M3093 (CAN1 in pBluescript II) into the HpaI site (position 2175) of pRS315 (ATCC 77144). The order of the major features in this plasmid is: LEU2 - CAN1 - f1 ori - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4. |
