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. 2020 Jun 19;10:9962. doi: 10.1038/s41598-020-66997-5

Table 1.

Plasmids and yeast strains used in this study. Genes from Aspergillus nidulans (An), Aspergillus clavatus (Ac), Clostridium acetobutylicum (Ca), Penicillium patulum (Pp), Saccharomyces cerevisiae (Sc), Salmonella enterica serovar typhimurium (St), Treponema denticola (Td) and codon-optimized genes (opt) are indicated by prefixes in superscript.

Plasmid Plasmid based on Relevant features Reference
pUG6-H pBR322, hphNT1, Ampr 23
pRS42K 2 µ, kanMX, Ampr 24
pRS72N 2 µ, natMX, Ampr 24
pRCC-K 2 µ, kanMX, Ampr, pROX3-optCas9-tCYC1, pSNR52-gRNA 25
pRS42K_prpE783Δ pJHV1 2 µ, kanMX, Ampr, pMET25, tCYC1, pTDH3-StoptprpEG783-tPGK1 This work
pJHV19 pRCC-K 2 µ, kanMX, Ampr, pROX3-optCas9-tCYC1, pSNR52-gRNA for SFA1 This work
pJHV54 pRCC-K 2 µ, kanMX, Ampr, pROX3-optCas9-tCYC1, pSNR52-gRNA for CIT3 This work
pJHV62 ColE1, AmpR,ConL3-pTDH3-Tdoptter-tADH1 –ConRE‘ This work
pJHV65 ConLS‘-pPGK1p-ScERG10-tVMA16 -ConR1-ConL1-pCCW12-Caopthbd-tIDP –ConR2-ConL2-pENO2-Caoptcrt-tPGK1 –ConR3-ConL3-pTDH3-Tdoptter-tADH1 –ConRE‘-natMX-LEU2 3Hom-KanR-ColE1-LEU 5Hom This work
pRS72N_ADY2 2 µ, natMX, Ampr, pHXT7-1–392-ScADY2-tCYC1 26
pRS72N_JEN1 2 µ, natMX, Ampr, pHXT7-1–392-ScJEN1-tCYC1 26
SiHV110 ConLS’-gfp-dropout-ConRE’-natMX-LEU2 3’Hom-KanR-ColE1-LEU 5’Hom This work (provided by Simon Harth)
pYTK3.41 ColE1, CamR, Tdoptter This work (provided by Fernando Garcés Daza)
pYTK3.43 ColE1, CamR, Caoptcrt This work (provided by Fernando Garcés Daza)
pYTK3.47 ColE1, CamR, ScERG10 This work (provided by Fernando Garcés Daza)
pYTK3.49 ColE1, CamR, Caopthbd This work (provided by Fernando Garcés Daza)
pYTK_Erg10 ColE1, AmpR,ConLS‘-pPGK1-ScERG10-tVMA16 -ConR1‘ This work (provided by Fernando Garcés Daza)
pYTK_Hbd ColE1, AmpR,ConL1-pCCW12-Caopthbd-tIDP –ConR2 This work (provided by Fernando Garcés Daza)
pYTK_Crt ColE1, AmpR,ConL2-pENO2-Caoptcrt-tPGK1 –ConR3 This work (provided by Fernando Garcés Daza)
pYTK01 ColE1, CamR, gfp-dropout 27
pYTK05 ColE1, CamR, ConL3 27
pYTK09 ColE1, CamR, TDH3p 27
pYTK53 ColE1, CamR, ADH1t 27
pYTK72 ColE1, CamR, ConRE 27
pYTK95 ColE1, CamR, AmpR-ColE1 27
pVS06 CEN6ARS4, kanMX, Ampr, pHXT7-1–392-ScERG10-tVMA16, pPGK1-Caopthbd-tEFM1, pTPI1-Caoptcrt-tYHI9, pPYK1-Tdoptter-tIDP1, pADH1-CaoptadhE2-tRPL3, pTDH3-EcopteutE-tRPL41B 22
pRS62H_ter 2 µ, natMX, Ampr, pHXT7-1–392-Tdoptter-tFBA1 21
pAB02 2 µ, natMX, Ampr, pROX3-optCas9-tCYC1, pSNR52-gRNA for POX1 This work (provided by Alexander Bissl)
pAB09 2 µ, natMX, Ampr, pROX3-optCas9-tCYC1, pSNR52-gRNA for CIT2 This work (provided by Alexander Bissl)
S. cerevisiae strain Parent strain Relevant features Reference
CEN.PK2-1C MATa leu2-3,112 ura3-52 trp1-289 his3-Δ1 MAL2–8c SUC2 28
JHY65 CEN.PK2-1C psfa1-sfa1Δ::pTDH3-StoptprpE-tSFA1 This work
JHY162 CEN.PK2-1C ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 14
JHY164 CEN.PK2-1C cit2Δ This work
JHY174 JHY164 cit2Δ cit3Δ This work
JHY175 CEN.PK2-1C cit3Δ This work
JHY179 JHY65 psfa1-sfa1Δ::pTDH3-StoptprpE-tSFA1 cit3Δ This work
JHY180 JHY179 psfa1-sfa1Δ::pTDH3-StoptprpE-tSFA1 cit3Δ cit2Δ This work
JHY185 JHY180 psfa1-sfa1Δ::pTDH3-StoptprpE-tSFA1 cit3Δ cit2Δ ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 This work
JHY194 JHY162 ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 leu2::pPGK1-ScERG10-tVMA16, pCCW12-Caopthbd-tIDP, pENO2-Caoptcrt-tPGK1, pTDH3-Tdoptter-tADH1, pTEF-natMX-tTEF This work
JHY196 CEN.PK2-1C pox1Δ This work
JHY197 JHY174 cit2Δ cit3Δ ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 This work
JHY211 JHY196 pox1Δ ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 This work
JHY212 JHY211 pox1Δ ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 leu2::pPGK1-ScERG10-tVMA16, pCCW12-Caopthbd-tIDP, pENO2-Caoptcrt-tPGK1, pTDH3-Tdoptter-tADH1, pTEF-natMX-tTEF This work
JHY218 JHY65 sfa1p-sfa1Δ::TDH3p-StoptprpE-SFA1t ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 This work
JHY229 JHY185 psfa1-sfa1Δ::pTDH3-StoptprpE-tSFA1 cit3Δ cit2Δ ura3::pPGK1-PpoptMSAS-tCYC1, pHXT7-1–392- AnoptnpgA-tFBA1, pFBA1-AcoptpatG-tADH1 pacs2-acs2Δ::pTEF-hphNT1-tCYC1 This work

Other abbreviations: hphNT1: hygromycin resistance; Ampr: ampicillin resistance; CamR: chloramphenicol resistance; KanR: kanamycin resistance; kanMX: geneticin resistance; natMX: clonat resistance. If not stated otherwise, promoters (p) were taken 1-500 bp upstream and terminators (t) 1-300 bp downstream of respective open reading frames.