Hi there,

These are strong constitutive promoters and terminators. Any combination should be OK... I have successfully expressed simultaneously 6 human proteins from genomic insertions using the following combinations: ProPGK1+terTDH1; ProTDH3+TerADH1; ProHHF2+TerSSA1; ProCCW12+TerENO1; ProTEF1+TerENO2; ProTEF2+TerPGK1. The most crucial point being to optimize sequences for expression in yeast.

You can use 2A sequences in between the the genes (polycistronically) and a single inducible promoter (pGal) terminator signal to reduce recombination events also if you add a drop out selection gene after the last 2A sequence then you can be sure that the whole polycistronic system works properly by inducing proteins in a dropout media.

Eg: gal1 promoter + RBS + GENE1-2A-gene2-2A....GeneN-2A-Ura3/His + strong terminator.

Different 2A sequences used in yeast successfully:

https://doi.org/10.1021/acssynbio.2c00506

Selecting the right promoter-terminator combinations for protein expression in Saccharomyces cerevisiae (baker's yeast) can depend on several factors, including the specific requirements of your experiment, the level of expression you need, and the characteristics of your protein of interest. Here are a few commonly used promoter-terminator combinations in S. cerevisiae:

It's important to note that the choice of promoter-terminator combination should be tailored to your specific needs. Factors to consider include the expression level required for your protein, whether you need inducible or constitutive expression, and any potential toxicity or growth effects associated with high-level expression. Additionally, some expression vectors and kits are commercially available that provide a range of promoter-terminator combinations for easy cloning and expression in S. cerevisiae.