Restriction digests of the clone give the following sizes (kb): HindIII--5.4, 2.4, 1.1; EcoRI--4.0, 3.6, 1.2; XhoI--8.9. 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). REP41X (ATCC 87605) and REP42X (ATCC 87607) are different in that REP41X contains the LEU2 marker and REP42X, ura4+. REP41X was constructed by deleting the ATG in the polylinker of the original REP41 plasmid. An XhoI linker was cloned into the blunted NdeI-BamHI cut REP41 plasmid, deleting SalI site. |
