Restriction digests of the clone give the following sizes (kb): HindIII--5.6, 2.3, 1.1; EcoRI--4.3, 3.5, 1.2; XhoI--9.0. REP3X (ATCC 87603) and REP4X (ATCC 87604) contain full strength nmt1 promoter whereas REP41X (ATCC 87605) and REP42X (ATCC 87607) with medium strength (nmt1*) and REP81X (ATCC 87606) and REP82X (ATCC 87608) with low strength (nmt1**) due to mutation in the TATA box by Basi et al. (Gene 123:131-136, 1993). REP3X (ATCC 87603) and REP4X (ATCC 87604) differ in that REP3X contains LEU2 (2.2 kb) and REP4X contains ura4+ (1.8 kb). REP3X was constructed by removing an ATG within the polylinker of an expression vector REP3, which contains the thiamine-inducible nmt1 promoter (J. Biol. Chem. 265:10857-10862, 1993), by insertion of a XhoI linker into Ball and SalI sites. The SalI site is preserved. |
