Restriction digests of the clone give the following sizes (kb): BglII--4.6; SalI/BglII--3.9, 0.7; EcoRI--4.6. The marker and promoter are flanked by the F4 primer site (5') and the R6 primer site (3'). The size of the F4/R6 PCR product should be 2005 nt. F4 primer sequence: 5'-(gene-specific sequence)GAATTCGAGCTCGTTTAAAC-3'. R6 primer sequence: 5'-(gene-specific sequence) CATTGTATATGAGATAGTTGA-3'. Contains a functional truncated (700 nt) ADH promoter. The promoter is not down regulated during the exponential phase. |
