Table 1 S. cerevisiae strains and plasmids used in this study
From: Heterologous biosynthesis and manipulation of crocetin in Saccharomyces cerevisiae
| Â | Description | Source |
|---|---|---|
Strain | ||
 CEN.PK2-1C | MATa, ura3-52, trp1-289, leu2-3,112, his3Δ1, MAL2-8C, SUC2 | EUROSCARF |
 SyBE_Sc0014CY06 | CEN.PK2-1C, Δgal1 Δgal7 Δgal10::HIS3, Δypl062w::KanMX, trp1::TRP1_T CYC1 -BtCrtI-P GAL10 -P GAL1 -PaCrtB-T PGK1 , leu2::LEU2_T CYC1 -BtCrtI-P GAL7 -T ACT1 -tHMG1-P GAL10 -P GAL1 -TmCrtE-T GPM1 , Δymrwdelta15::P UAS-GAL1 -PaCrtY-T ADH1 , Δynrcdelta9:: P UAS-GAL1 -PaCrtY-T ADH1 | This lab |
 SyBE_Sc0123Z001 | SyBE_SC0014CY06, Δho::P GAL1 -Aa_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z002 | SyBE_SC0014CY06, Δho::P GAL1 -As_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z003 | SyBE_SC0014CY06, Δho::P GAL1 -Eu_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z004 | SyBE_SC0014CY06, Δho::P GAL1 -Pa_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z005 | SyBE_SC0014CY06, Δho::P GAL1 -Ps_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z006 | SyBE_SC0014CY06, Δho::P GAL1 -Ss_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z007 | SyBE_SC0014CY06, Δho::P GAL1 -B.SD_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z008 | SyBE_SC0014CY06, Δho::P GAL1 -B.DC_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Z009 | SyBE_SC0014CY06, Δho::P GAL1 -Hp_CrtZ-T HIS5 -URA3 | This study |
 SyBE_Sc0123Cz10 | SyBE_SC0014CY06, Δho::P GAL1 -Aa_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz11 | SyBE_SC0014CY06, Δho::P GAL1 -As_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz12 | SyBE_SC0014CY06, Δho::P GAL1 -Eu_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz13 | SyBE_SC0014CY06, Δho::P GAL1 -Pa_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz14 | SyBE_SC0014CY06, Δho::P GAL1 -Ps_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz15 | SyBE_SC0014CY06, Δho::P GAL1 -Ss_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz16 | SyBE_SC0014CY06, Δho::P GAL1 -B.SD_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz17 | SyBE_SC0014CY06, Δho::P GAL1 -B.DC_CrtZ-T HIS5 | This study |
 SyBE_Sc0123Cz18 | SyBE_SC0014CY06, Δho::P GAL1 -Hp_CrtZ-T HIS5 | This study |
 SyBE_Sc0123C001 | SyBE_Sc0123Cz10 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C002 | SyBE_Sc0123Cz10 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C003 | SyBE_Sc0123Cz10 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C004 | SyBE_Sc0123Cz10 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C005 | SyBE_Sc0123Cz11 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C006 | SyBE_Sc0123Cz11 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C007 | SyBE_Sc0123Cz11 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C008 | SyBE_Sc0123Cz11 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C009 | SyBE_Sc0123Cz12 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C010 | SyBE_Sc0123Cz12 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C011 | SyBE_Sc0123Cz12 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C012 | SyBE_Sc0123Cz12 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C013 | SyBE_Sc0123Cz13 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C014 | SyBE_Sc0123Cz13 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C015 | SyBE_Sc0123Cz13 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C016 | SyBE_Sc0123Cz13 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C017 | SyBE_Sc0123Cz14 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C018 | SyBE_Sc0123Cz14 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C019 | SyBE_Sc0123Cz14 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C020 | SyBE_Sc0123Cz14 with pRS416-C-04(pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C021 | SyBE_Sc0123Cz15 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C022 | SyBE_Sc0123Cz15 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C023 | SyBE_Sc0123Cz15 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C024 | SyBE_Sc0123Cz15 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C025 | SyBE_Sc0123Cz16 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C026 | SyBE_Sc0123Cz16 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C027 | SyBE_Sc0123Cz16 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C028 | SyBE_Sc0123Cz16 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C029 | SyBE_Sc0123Cz17 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C030 | SyBE_Sc0123Cz17 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C031 | SyBE_Sc0123Cz17 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C032 | SyBE_Sc0123Cz17 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C033 | SyBE_Sc0123Cz18 with pRS416-C-01 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 ) | This study |
 SyBE_Sc0123C034 | SyBE_Sc0123Cz18 with pRS416-C-02 (pRS416-T HIS5 -P GAL10 -CCD3-T TEF2 ) | This study |
 SyBE_Sc0123C035 | SyBE_Sc0123Cz18 with pRS416-C-03 (pRS416-T HIS5 -P GAL10 -ZCD-T TEF2 ) | This study |
 SyBE_Sc0123C036 | SyBE_Sc0123Cz18 with pRS416-C-04 (pRS416-T HIS5 -P GAL10 -ZCD1-T TEF2 ) | This study |
 SyBE_Sc0123C048 | SyBE_Sc0123Cz14 with pRS416-A-02 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 -P GAL7 -ALD6-T PGI1 ) | This study |
 SyBE_Sc0123C049 | SyBE_Sc0123Cz14 with pRS416-A-03 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 -P GAL7 - Bix_ALD-T PGI1 ) | This study |
 SyBE_Sc0123C050 | SyBE_Sc0123Cz14 with pRS416-A-04 (pRS416-T HIS5 -P GAL10 -CCD2-T TEF2 -P GAL7 - Syn_ALD-T PGI1 ) | This study |
 SyBE_Sc0123C053 | SyBE_Sc0123Cz14 with pRS426-A-02 (pRS426-T HIS5 -P GAL10 -CCD2-T TEF2 -P GAL7 - Syn_ALD-T PGI1 ) | This study |
Plasmid | ||
 pJET1.2 | Blunt Cloning vector, resistant to ampicillin | Thermo scientific |
 pUC57-Simple | Blunt Cloning vector, resistant to ampicillin | GenScript |
 pRS416 | Single copy plasmid in S.cerevisiae with URA3 and Ampr marker | This Lab |
 pRS426 | Multiple copy plasmid in S.cerevisiae with URA3 and Ampr marker | This Lab |
 pRS425 K | Multiple copy plasmid in S.cerevisiae with LEU2 and KanMX marker | This Lab |
 pUC57-Simple-01 | CrtZ from Agrobacterium aurantiacum (Aa_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-02 | CrtZ from Alcaligenes sp. PC-1 (As_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-03 | CrtZ from Erwinia uredovora (Eu_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-04 | CrtZ from Pantoea agglomerans (Pa_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-05 | CrtZ from Pantoea stewartii (Ps_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-06 | CrtZ from Sulfolobus solfataricus P2 (Ss_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-07 | CrtZ from Brevundimonas sp. SD212 (B.SD_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-08 | CrtZ from Brevundimonas vesicularis DC263 (B.DC_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-09 | CrtZ from Haematococcus pluvialis (Hp_CrtZ) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-10 | CCD2 from Crocus was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-11 | CCD3 from Crocus was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-12 | ZCD from Crocus was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-13 | ZCD1 from Crocus was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-14 | ALD6 from S. cerevisiae was cloned into pUC57-Simple | This study |
 pUC57-Simple-15 | ALD from Bixa orellana (Bix_ALD) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pUC57-Simple-16 | ALD from Synechocystis sp. PCC6803 (Syn_ALD) was codon optimized, synthesized and cloned into pUC57-Simple | This study |
 pJET1.2-Z-01 | The cassette ho_F-P GAL1 -T HIS5 -URA3-ho_R was cloned and inserted into the pJET1.2 | This study |
 pJET1.2-Z-02 | Aa_CrtZ was digested from pUC57-Simple-01 by BsaI and inserted into the same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-03 | As_CrtZ was digested from pUC57-Simple-02 by BsaI and inserted into the same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-04 | Eu_CrtZ was digested from pUC57-Simple-03 by BsaI and inserted into the same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-05 | Pa_CrtZ was digested from pUC57-Simple-04 by BsaI and inserted into the same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-06 | Ps_CrtZ was digested from pUC57-Simple-05 by BsaI and inserted into the same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-07 | Ss_CrtZ was digested from pUC57-Simple-06 by BsaI and inserted into the Same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-08 | B.SD_CrtZ was digested from pUC57-Simple-07 by BsaI and inserted into the Same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-09 | B.DC_CrtZ was digested from pUC57-Simple-08 by BsaI and inserted into the Same site of pJET1.2-Z-01 | This study |
 pJET1.2-Z-10 | Hp_CrtZ was digested from pUC57-Simple-09 by BsaI and inserted into the Same site of pJET1.2-Z-01 | This study |
 pRS416-C-01 | The cassette T HIS5 -P GAL10 -CCD2-T TEF2 was cloned and inserted into the NotI site of pRS416 | This study |
 pRS416-C-02 | The cassette T HIS5 -P GAL10 -CCD3-T TEF2 was cloned and inserted into the NotI site of pRS416 | This study |
 pRS416-C-03 | The cassette T HIS5 -P GAL10 -ZCD-T TEF2 was cloned and inserted into the NotI site of pRS416 | This study |
 pRS416-C-04 | The cassette T HIS5 -P GAL10 -ZCD1-T TEF2 was cloned and inserted into the NotI site of pRS416 | This study |
 pRS425 K-A-01 | The cassette T TEF2 -P GAL7 -T PGI1 was cloned and inserted into the PstI/BamHI site of pRS425 K | This study |
 pRS425 K-A-02 | ALD6 was digested from pUC57-Simple-14 by BsaI and inserted into the same site of pRS425 K-A-01 | This study |
 pRS425 K-A-03 | Bix_ALD was digested from pUC57-Simple-15 by BsaI and inserted into the same site of pRS425 K-A-01 | This study |
 pRS425 K-A-04 | Syn_ALD was digested from pUC57-Simple-16 by BsaI and inserted into the same site of pRS425 K-A-01 | This study |
 pRS416-A-01 | The cassette T HIS5 –T PGI1 was cloned and inserted into the XhoI/SacI site of pRS416 | This study |
 pRS416-A-02 | The cassette T HIS5 -P GAL10 -CCD2-T TEF2 (digested from pRS416-C-01 by NotI), the cassette T TEF2 -P GAL7 -ALD6-T PGI1 (digested from pRS425 K-A-02 by PstI/BamHI) and plasmid pRS416-A-01 (digested by BamHI) were assembled based on RADOM method | This study |
 pRS416-A-03 | The cassette T HIS5 -P GAL10 -CCD2-T TEF2 (digested from pRS416-C-01 by NotI), the cassette T TEF2 -P GAL7 -Bix_ALD-T PGI1 (digested from pRS425 K-A-03 by PstI/BamHI) and plasmid pRS416-A-01 (digested by BamHI) were assembled based on RADOM method | This study |
 pRS416-A-04 | The cassette T HIS5 -P GAL10 -CCD2-T TEF2 (digested from pRS416-C-01 by NotI), the cassette T TEF2 -P GAL7 -Syn_ALD-T PGI1 (digested from pRS425 K-A-04 by PstI/BamHI) and plasmid pRS416-A-01 (digested by BamHI) were assembled based on RADOM method | This study |
 pRS426-A-01 | The cassette T HIS5 –T PGI1 was cloned and inserted into the XhoI/SacI site of pRS426 | This study |
 pRS426-A-02 | The cassette T HIS5 -P GAL10 -CCD2-T TEF2 (digested from pRS416-C-01 by NotI), the cassette T TEF2 -P GAL7 -Syn_ALD-T PGI1 (digested from pRS425 K-A-04 by PstI/BamHI) and plasmid pRS426-A-01 (digested by BamHI) were assembled based on RADOM method | This study |