Synthetic biology and metabolic engineering enable generation of novel cell factories that efficiently convert renewable feedstocks into biofuels, bulk and fine chemicals, thus creating the basis for biosustainable economy independent on fossil resources. While over a hundred proof-of-concept chemicals have been made in yeast, only a very small fraction of those has reached commercial-scale production so far. The limiting factor is the high research cost associated with development of a robust cell factory that can produce the desired chemical at high titer, rate and yield. Synthetic biology has the potential to bring down this cost by improving our ability to predictably engineer biological systems. This review highlights synthetic biology applications for design, assembly and optimization of non-native biochemical pathways in baker's yeast Saccharomyces cerevisiae. We describe computational tools for prediction of biochemical pathways, molecular biology methods for assembly of DNA parts into pathways and for introducing the pathways into the host, and finally approaches for optimizing performance of the introduced pathways.This article is protected by copyright. All rights reserved.