Restriction digests of the clone give the following sizes (kb): EcoRI--5.0; BamHI--5.0, PvuII--4.4, 0.5. One of a series of pBluescript-based centromere vectors (ATCC 77142-77145, 77157-77158) differing in the yeast selectable marker gene. YC-type centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ') peptide. Useful in plasmid shuffle experiments. pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and f1 origin of pBS(+). A fragment (1.112 kb) containing the URA3 gene was inserted into the NdeI site and a cassette containing CEN6 and the ARS associated with histone 4 (ARSH4) was inserted into the AatII site of pRSS56. All ends were blunted. The order of the major features in this plasmid is: URA3 - f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4. |
