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Table 4 Strains and plasmids used in this study

From: ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production

 

Description

Reference

Strain

7D

S. cerevisiae CEN.PK113-7D. MATa; URA3; TRP1; LEU2; HIS3; MAL2-8c; SUC2

Euroscarf

7D-atp2

7D with genomic integration of the T. reesei ATPase β subunit gene atp2 (pTEF1-atp2-tCYC1) at X2 locus

This study

7D-atpAGD (2A)

7D with genomic integration of the E. coli ATPase α, β and γ subunit genes atpAGD joined via viral 2A peptides at X2 locus

This study

5D

S. cerevisiae CEN.PK113-5D. MATa; ura3-52; TRP1; LEU2; HIS3; MAL2-8c; SUC2

Euroscarf

5D (E.V.)

5D transformed with the empty vector pRSII326

This study

5D-atp2

5D transformed with pRSII326-atp2

This study

5D-atpAGD

5D transformed with pRSII326-atpAGD

This study

5D-atpAGD (2A)

5D transformed with pRSII326-atpAGD (2A)

This study

Plasmids

pCfB2312

Cas9 Expression Vector

[35]

pCfB2899

EasyClone-MarkerFree Integrative Vector for insertion into Chromosome X: 194944...195980 (X2 locus)

[35]

pCfB3020

EasyClone-MarkerFree gRNA Helper Vector for insertion into X2 locus

[35]

pCfB2899-atp2

pCfB2899 carrying the T. reesei ATPase β subunit gene atp2 (pTEF1-atp2-tCYC1)

This study

pCfB2899-atpAGD (2A)

pCfB2899 carrying the E. coli ATPase α, β and γ subunit genes atpAGD joined with viral 2A peptide sequences under the control of TEF1 promoter

This study

pRSII326

Shuttle vector with URA3 marker and 2µ ORI-STB. Addgene ID: 35469

[36]

pRSII326-atp2

pRSII326 with T. reesei ATPase β subunit gene atp2 (pTEF1-atp2-tCYC1)

This study

pRSII326-atpAGD

pRSII326 with E. coli ATPase α subunit gene atpA (pTPI-atpA-tCPS1), subunit β gene atpD (pTEF1-atpD-tPRM9) and γ subunit gene atpG (pGPM1-atpG-tHIS5)

This study

pRSII326-atpAGD (2A)

pRSII326 with E. coli ATPase α, β and γ subunit genes atpAGD joined with viral 2A peptide sequences under the control of TEF1 promoter

This study