Size (kb): 4.8340001106262210
Vector: pFa6a-HISMX6-pADH1-3HA (plasmid)
Construct size (kb): 4.834000110626221
Features: marker(s): ampR, HIS
tag (N-terminal): HA tag
Restriction digests of the clone give the following sizes (kb): BglII--4.8; EcoRI--4.8; BglII/BamHI--4.0, 0.8.
The marker, promoter and 3HA tag are flanked by the F4 primer site (5') and the R3 primer site (3'). The size of F4/R3 PCR product is 1969 nt.
R3 primer sequence: 5'(gene-specific sequence)GCACTGAGCAGCGTAATCTG-3'. The gene-specific sequence of these primers are chosen to correspond to the reverse complement of the N-terminal codons of the target gene. The sequences shown here are the reverse 3'.
Contains a functional truncated (700 nt) ADH promoter. The promoter is not down regulated during the exponential phase.
DeMarini DJ, et al. Constitutive promoter modules for PCR-based gene modification in Saccharomyces cerevisiae. Yeast 18: 723-728, 2001. PubMed: 11378899