PCR primers for amplification of the modules in Figure 3.

1The forward primer is identical for all modules described here and is the same as for previously described modules containing nmt1-derived promoters [26]. The gene-specific portion of the primer is typically chosen to correspond to sequences 100–200 bp upstream of the start codon. A web-based tool for automated primer design is available for these primers [35].
2A 25-mer universal sequence is used to anneal to Purg1 due to the AT-rich nature of this sequence; the gene-specific portion is therefore reduced to 75 bp for 100-mer primers, which does not seem to affect targeting efficiency. The complement start codon is indicated in italic. For regulated expression of full length proteins, the gene-specific portion of the primer corresponds to the complement of the N-terminal codons of the target gene (without start codon).
3The reading frames of the tag sequences are indicated. These primers are the same as for the corresponding modules containing nmt1-derived promoters [26]. For N-terminal tagging of full-length proteins, the gene-specific portion of the primer corresponds to the complement of the N-terminal codons of the target gene (including start codon). Note that the 3′ portions of these primers are specific to the tags and correspond to the complement of the C-terminal tag codons (without stop codon). A web-based tool for automated primer design is available for these primers [35].