Ez transformation kit

S. cerevisiae strain CEN.PK2-1Dα was used as the base strain for in this work with the exception of the evolution of yeTadA1.0 adenosine deaminase, which was performed in W303α as noted in Supplementary Table 1. Characterization of TadA and engineered yeTadA1.0 was performed in a CEN.PK2α as noted in the figures and Supplementary Table 1. All yeast plasmid transformations were performed using EZ transformation kit from Zymo Research according to manufacturer’s instructions, unless otherwise specified. Cloning was performed using a combination of Gibson assembly and DNA gap repair in the yeast strains specified in Supplementary Table 1. Scar-free yeast integrations were carried out as previously described² , by co-transformation of gRNA/Cas9 expression plasmid targeting the edit site, along with repair DNA to be integrated genomically. Briefly, expression cassettes or gene modifications were amplified using primers with 15–40 bp overhangs for gap repair in yeast. Each assembly was flanked by an integration homology region of 30–50 base pairs. gRNAs were generated at a target region such that integration of the desired modification would result in disruption of the gRNA binding site. Typically, 100 ng of purified PCR product was introduced with 300 ng of gRNA plasmid using the Zymo EZ Kit for transformation.