Table 1.

Oligonucleotides used to generate mutant C. parapsilosis isolates carrying TAC1^L518F and FKS1^E1393G.

Production of the guide RNA The protospacer sequence/PAM site is indicated, followed by the 23-mers (TOP and BOT oligos) for generating the guide RNA. Overhangs are highlighted in bold
TAC1-g1 (sequence)	5′-GTGGCTGATGAGGCATTACT/TGG-3′
TAC1-g1-TOP	5′-CCAGTGGCTGATGAGGCATTACT-3′	Annealing of the 23-mer to produce the guide RNA that was cloned into SapI-digested pCP-tRNA
TAC1-g1-BOT	5′-AACAGTAATGCCTCATCAGCCAC-3′
FKS1-g1 (sequence)	5′-AGTTGATTGAAAGAGGTGTG/TGG-3′
FKS1-g1-TOP	5′-CCAAGTTGATTGAAAGAGGTGTG-3′	Annealing of the 23-mer to produce the guide RNA that was cloned into SapI-digested pCP-tRNA
FKS1-g1-BOT	5′-AACCACACCTCTTTCAATCAACT-3′
Synthesis of RTs The sequence of each RT is indicated, followed by the long oligos (TOP and BOT) used for primer extension. The SNPs are highlighted in bold, and described on the right (the numbering refers to the ORF); syn = synonymous
TAC1-RT-1 (sequence)	5′-GGCGACGAATTGGATCGTCAAATGTCGATTGCAGTGGCTGATGAGGCATTGTTTGGAGATCCAGCACTACCACTCAGTTTTCGATTGTTGAAAAAGTTGA-3′	SNPs introduced: A1551G [syn SNP]; C1552T [non syn SNP]
TAC1-RT-1-TOP	5′-GGCGACGAATTGGATCGTCAAATGTCGATTGCAGTGGCTGATGAGGCATTGTTTGGAGAT-3′
TAC1-RT-1-BOT	5′-TCAACTTTTTCAACAATCGAAAACTGAGTGGTAGTGCTGGATCTCCAAACAATGCCTCAT-3′
FKS1-RT-1 (sequence)	5′-TCTTCATTTCGTTCATTCCATTGGTTGTTCAAGGGTTGATTGAAAGAGGAGTCTGGAAAGCTTGTCAAAGATTTGTTAGACATTTCATTTCGTTGTCACC-3′	A4178G [non syn SNP]; G4197C [syn SNP]; T4194A [syn SNP]
FKS1-RT-1-TOP	5′-TCTTCATTTCGTTCATTCCATTGGTTGTTCAAGGGTTGATTGAAAGAGGAGTCTGGAAAG-3′
FKS1-RT-1-BOT	5′-GGTGACAACGAAATGAAATGTCTAACAAATCTTTGACAAGCTTTCCAGACTCCTCTTTCA-3′
Sequencing of mutated loci
sTAC1-F	5′-GGTATGCTCAGGAGATTGGA-3′	Sequencing of TAC1 edited site
sTAC1-R	5′-ATAGTTCCACGTTCAGGCTC-3′
sFKS1-F	5′-CGGACATCCTGGTTTCCATA-3′	Sequencing of FKS1 edited site
sFKS1-R	5′-CAATGAAGACAACGAAGCCC-3′