First question is; how to set up a good antibiotic-free cloning system
for yeast. So, yeast minicircle plasmid with allelopathic toxin,
perhaps? I recall these occur naturally, plasmids that kill cells not
containing the toxin, so they prevent plasmid loss in the population.
Doubt that the toxin is harmful to humans.
There is a long history of using auxotrophic mutants for cloning in S. cerevisiae, integrative, centromeric and high copy replicative vectors (URA3 and LEU2, other markers, complementing appropriate mutant strains). This wheel should already have been invented, not sure if there is a central database for these vectors/strains unfortunately but a lot of yeast labs around the world in academia to ask.
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