The pPICHOLI vectors are designed for heterologous gene expression in the yeast P. pastoris and in the prokaryote E. coli. The vectors contain an inducible (yeast) alcohol oxidase (AOX) promoter and an E. coli T7 promoter. The vectors also contain sequences allowing autonomous replication both in P. pastoris and E. coli. Due to these components, vector linearization is no longer required and small amounts of DNA are sufficient to successfully transform P. pastoris. The integrated PARS sequence enables simple recovery of plasmids from yeast. Time-consuming subcloning into several expression vectors, including testing for a successful gene expression, is no longer necessary. A multiple cloning site enables convenient ligation of DNA fragments into the vectors.