. 2018 Feb 9;18(3):foy012. doi: 10.1093/femsyr/foy012

Table 2.

Plasmids used in this study.

Name (Addgene Plasmid #)	Relevant characteristics	Origin
pYTK079	Template for hph (Hyg^R) open reading frame	Lee et al. (2015)
pUD423	Template for AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t cassette	Gorter de Vries et al. (2017)
pUD527	ori kan^R SHRA AgTEF1p-amdS-AgTEF1t SHRB	GeneArt
pUD530	ori kan^R panARS(OPT)	GeneArt
pUD531	ori kan^R SHRC ScTDH3p-^{BsaI BsaI}-ScCYC1t SHRI	GeneArt
pUD532	ori kan^R SHRI ori bla SHRA	GeneArt
pUD540	ori amp^R ^BsaIHH-gRNA_OpADE2-HDV^BsaI	GeneArt
pUD555	ori amp^R ^BsaIHH-gRNA_KlADE2-HDV^BsaI	GeneArt
pUD602	ori amp^R ^BsaIHH-gRNA_OpKU80-HDV^BsaI	GeneArt
pUD750	ori amp^R ^BsaIHH-gRNA_OpADE2-HDV-HH-gRNA_OpYNR1-HDV^BsaI	This study
pUDP002 (plasmid #103872)	ori amp^R panARS(OPT) AgTEF1p-hph-AgTEF1t ScTDH3p^{BsaI BsaI}ScCYC1t AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t	This study
pUDP013 (plasmid #103873)	ori amp^R panARS(OPT) AgTEF1p-hph-AgTEF1t ScTDH3p-HH-gRNA_OpADE2-HDV- ScCYC1t AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t	This study
pUDP025 (plasmid #103874)	ori amp^R panARS(OPT) AgTEF1p-hph-AgTEF1t ScTDH3p-HH-gRNA_KlADE2-HDV- ScCYC1t AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t	This study
pUDP046 (plasmid #107062)	ori amp^R panARS(OPT) AgTEF1p-hph-AgTEF1t ScTDH3p-HH-gRNA_OpKU80-HDV- ScCYC1t AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t	This study
pUDP082 (plasmid #103875)	ori amp^R panARS(OPT) AgTEF1p-hph-AgTEF1t ScTDH3p-HH-gRNA_KmADE2-HDV- ScCYC1t AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t	This study
pUDP123 (plasmid # 107269)	ori amp^R panARS(OPT) AgTEF1p-hph-AgTEF1t ScTDH3p-HH-gRNA_OpADE2-HDV-HH-gRNA_OpYNR1-HDV-ScCYC1t AaTEF1p-Spcas9^{D147Y P411T}-ScPHO5t	This study

Restriction enzyme sites are indicated in superscript and gRNA target sequences are indicated in subscript. SHRs represent specify synthetic homologous recombination sequences used for plasmid assembly. Aa: Arxula adeninivorans; Sp: Streptococcus pyogenes; Ag: Ashbya gossypii; Sc: Saccharomyces cerevisiae; Op: Ogataea (para)polymorpha; Kl: Kluyveromyces lactis; Km: Kluyveromyces marxianus; HH: hammerhead ribozyme; HDV: hepatitis delta virus ribozyme. pUDP013 (gRNA_OpADE2) targets ADE2 in both O. polymorpha and O. parapolymorpha. The addgene plasmid code (when relevant) is indicated next to the plasmid name between brackets.