Figure 1. FIGURE 1: Schematic representation of the constructed vectors.

The pRA vectors (A) are yeast centromeric shuttle plasmids that carry an auxotrophic marker (URA3, TRP1 or LEU2 gene) for selection in yeast and the ampicillin (Amp) resistance gene for selection in E. coli. The gene of interest can be cloned into the multiple cloning site (MCS) under control of either the inducible GAL1 promoter (3 different strengths) or the constitutive TEF promoter. The expressed protein will bear any of the four (Flag, Myc, HA or V5) N-terminal epitope tags allowing easy detection. All restriction sites indicated are unique. (B) High copy (2 μm) yeast shuttle plasmids carrying the HIS3 auxotrophic marker and expressing either 6xHis-Myc tagged ubiquitin (Ub) or SUMO (Smt3) under control of the copper inducible promoter (CUP). CTA1 and Cyc1 term (inator), transcription termination sequences derived from the yeast Catalase A and Cytochrome c1 gene, respectively.