Restriction digests of the clone give the following sizes (kb): HindIII--4.8, 2.2, 1.4, 1.2; BamHI--9.6; NotI--9.6; ClaI--9.6. In S. cerevisiae, requires a host expressing FLP such as YPH485 or YPH494, or a host modified by pFV17 (ATCC 77170). When propagated in a strain carrying an integrated copy of GAL10-FLP, the copy number is 10-20 without induction and 200-400 when induced by galactose. YE-type shuttle vector permitting FLP-driven high copy number. Constructed by inserting a BalI/EcoRV fragment of the 2 micron circle (interrupting FLP) into the SmaI site of pHSS6 which had been modified by the addition of a TRP1 gene. The order of the major features in this plasmid is: REP3 - D - REP1 - REP2 - TRP1 - MCS (NotI - BamHI/NotI) - kanR. |
