Table 2.
Plasmids used throughout the study
| Name | Relevant genotype | Source | Addgene ID#a |
|---|---|---|---|
| pCT | ori amp r ARS4 CEN6 LEU2 AaTEF1p-Spcas9 D147Y P411T-ScPHO5 | [36] | |
| pMEL15 | ori amp r 2 μm natNT2 SNR52p-gRNACAN1-SUP4t | [15] | |
| pROS12 | ori amp r 2 μm hphNT1 gRNA CAN1 gRNA ADE2 | [15] | |
| pUC19 | ori amp r LacZ multiple cloning site | [68] | |
| pUD423 | ori amp r AaTEF1p-Spcas9 D147Y P411T-ScPHO5t | GeneArt™ | |
| pUD526 | ori amp r NotI HRL AaTEF1p-Spcas9 D147Y P411T-ScPHO5t natNT2 HRMNotI | GeneArt™ | |
| pUD527 | ori kan r SHRA AgTEF1p-amdS-AgTEF1t SHRB | GeneArt™ | |
| pUD528 | ori kan r SHRB AaTEF1p-Spcas9 D147Y P411T-PHO5t | GeneArt™ | |
| pUD530 | ori kan r SHRB panARSopt SHRC | GeneArt™ | |
| pUD531 | ori kan r SHRC TDH3p BsaI BsaI CYC1t SHRI | GeneArt™ | |
| pUD532 | ori kan r SHRI bla ori SHRA | GeneArt™ | |
| pUD536 | ori amp r BsaI gRNABsaISeILV6 | GeneArt™ | |
| pUD573 | ori amp r BsaI gRNASeATF1 gRNABsaISeATF2 | GeneArt™ | |
| pUDP003 | ori amp r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p BsaI BsaI CYC1t | This study | 101164 |
| pUDP004 | ori amp r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p BsaI BsaI CYC1t AaTEF1p-Spcas9 D147Y P411T-ScPHO5t | This study | 101165 |
| pUDP010 | ori amp r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p-HH-gRNASeILV6-HDV-CYC1t | This study | 101166 |
| pUDP012 | ori amp r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p-HH-gRNASeILV6-HDV-CYC1t AaTEF1p-Spcas9 D147Y P411T-ScPHO5t | This study | 101167 |
| pUDP044 |
ori amp
r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p-HH-gRNASeATF1 -HDV-HH-gRNA
SeATF2-HDV-CYC1t
AaTEF1p-Spcas9 D147Y P411T-ScPHO5t |
This study | 101168 |
| pUDR107 | ori amp r 2 μm hphNT1 gRNAURA3 | This study |
HRL and HRM indicate the left and right homology arms for integration on the SPR3 locus, SHR stands for synthetic homologous recombination sequence and enzyme digestion sites are indicated in superscript