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Table 2 Plasmids used throughout the study

From: CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces pastorianus

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-gRNA CAN1 -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

GeneArtâ„¢

 

pUD526

ori amp r NotI HRL AaTEF1p-Spcas9 D147Y P411T-ScPHO5t natNT2 HRMNotI

GeneArtâ„¢

 

pUD527

ori kan r SHRA AgTEF1p-amdS-AgTEF1t SHRB

GeneArtâ„¢

 

pUD528

ori kan r SHRB AaTEF1p-Spcas9 D147Y P411T-PHO5t

GeneArtâ„¢

 

pUD530

ori kan r SHRB panARSopt SHRC

GeneArtâ„¢

 

pUD531

ori kan r SHRC TDH3p BsaI BsaI CYC1t SHRI

GeneArtâ„¢

 

pUD532

ori kan r SHRI bla ori SHRA

GeneArtâ„¢

 

pUD536

ori amp r BsaI gRNA BsaISeILV6

GeneArtâ„¢

 

pUD573

ori amp r BsaI gRNASeATF1 gRNA BsaISeATF2

GeneArtâ„¢

 

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-gRNA SeILV6 -HDV-CYC1t

This study

101166

pUDP012

ori amp r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p-HH-gRNA SeILV6 -HDV-CYC1t AaTEF1p-Spcas9 D147Y P411T-ScPHO5t

This study

101167

pUDP044

ori amp r panARSopt AgTEF1p-amdS-AgTEF1t TDH3p-HH-gRNA SeATF1 -HDV-HH-gRNA SeATF2-HDV-CYC1t

AaTEF1p-Spcas9 D147Y P411T-ScPHO5t

This study

101168

pUDR107

ori amp r 2 μm hphNT1 gRNA URA3

This study

 
  1. 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
  2. a https://www.addgene.org/