show Abstracthide AbstractThree temperature sensitive (TS) mutants in the budding yeast strain S288C were backcrossed into three natural yeast isolates. For each TS mutant - genetic background combination, three spontaneous suppressor mutants were isolated that could suppress the temperature sensitivity, and sequenced whole-genome. Provided are whole-genome sequencing data of the 27 suppressor mutants as well as the 9 parental strains.