Integrative modules for efficient genome engineering in yeast.

We present a set of vectors containing integrative modules for efficient genome integration into the commonly used selection marker loci of the yeast . A fragment for genome integration is generated via PCR with a unique set of short primers and integrated into , , , and loci. The desired level of expression can be achieved by using constitutive (, ), inducible (, ), and daughter-specific () promoters available in the modules. The reduced size of the integrative module compared to conventional integrative plasmids allows efficient integration of multiple fragments. We demonstrate the efficiency of this tool by simultaneously tagging markers of the nucleus, vacuole, actin, and peroxisomes with genomically integrated fluorophores. Improved integration of our new pDK plasmid series allows stable introduction of several genes and can be used for multi-color imaging. New bidirectional promoters (, , and ) allow tractable metabolic engineering.