Restriction digests of the clone give the following sizes (kb): KpnI--8.0; SacI--8.0; BamHI--8.0. Differs from pU2X1 (ATCC 77466) only in the orientation of the hsp70 promoter-terminator cassette. The sequence conferring resistance to carboxin (cbxR) is a mutation of an iron-sulfur protein subunit of the Ustilago maydis succinate dehydrogenase gene. A high-copy-number cloning vector permitting visual detection of recombinants using insertional inactivation of the Rhodococcus gene for indigo production. Includes priming sites useful for sequencing. Shuttle expression vector using Ustilago maydis promoters gap (glyceraldehyde phosphate dehydrogenase) or hsp70 to drive expression of the cloned insert in the BamHI or KpnI cloning site respectively. Constructed from pUXV3 (ATCC 77465) by inserting a 2.1 kb hsp70 promoter and termination cassette into the KpnI site, creating a unique KpnI cloning site between the promoter and terminator. The order of the major features in the plasmid is: Ustilago maydis ARS - ampR - pMB1 ori - 5' lacZ fragment - EcoRI - SacI <- hsp70 terminator - KpnI <- hsp70 promoter - gap promoter -> BamHI <- cbxR - 3' lacZ fragment. |
