The
order of the major features in this plasmid is:ori(f1) - lacZ - T7 promoter -
MCS (KpnI-SacI) - T3 promoter - lacI - ori(pMB1) - ampR - ori (2 micron) -
URA3. Contains the REP3 and FRT sequences necessary for high copy propagation
in yeast. In S. cerevisiae, the copy number is about 20 per haploid cell. In
non-selective growth, plasmids are lost through mitotic segregation at rates in
the range of 4.4 + or - 1.4% of progeny per doubling. YE-type shuttle vector
encoding the beta-galactosidase alpha peptide, permitting blue-white visual
detection. Can be used to produce ssDNA. Contains promoters for in vitro RNA
synthesis and priming sites useful for sequencing. Restriction digests of the
clone give the following sizes (kb): BamHI--5.8; EcoRI--5.8; PstI--4.0, 1.7;
PvuII--5.2, 0.54. (ATCC staff) Medium is 1227 LB plus ampicillin. NCBI gi: 416322
Hosts: E.coli DH5alpha, Saccharomyces cerevisiae, E.coli.