TABLE 1.
Bacterial strains and plasmids used in this study.
| Strains | Relevant genotype | Source |
| E. coli NZY5α | fhuA2Δ(argF–lacZ)U169 phoA glnV44Φ80 Δ(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 | NZYTech (MB00401) |
| E. coli BL21(DE3) | F– ompT gal dcm lon hsdSB(rB- mB-) λ(DE3 ∗lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | NZYTech (MB006) |
| E. coli BL21(DE3) ΔlacZ | F– ompT gal dcm lon hsdSB(rB- mB-) λ(DE3 ∗lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) ΔlacZ | This study |
| Plasmids | Construct | Source |
| pCas9Cr4 | p15A ori, Cas9 from Streptococcus pyogenes under control of PTET, constitutive tetR, CmR | Addgene (62655) |
| pKDsgRNA-p15A | SC101 ori, sgRNA targeting p15A ori under control of PTET,λred under control of ParaB, araC, SpecR | Addgene (62656) |
| pKDsgRNA-LacZ | pKDsgRNA-p15A with sgRNA targeting lacZ instead of p15A ori | This study |
| pRSFduet-1 | RSF1030 ori, lacI, double PT7lac, KanR | Novagen |
| pACYCduet-1 | p15A ori, lacI, double PT7lac, CmR | Novagen |
| pUC57_COMT | pMB1 ori, AmpR; pUC57 carrying codon-optimized COMT from Arabidopsis thaliana | NZYTech |
| pRSFduet_TAL | pRSFduet-1 carrying codon-optimized TAL from Rhodotorula glutinis | Rodrigues et al., 2015a |
| pCDFduet_TAL | CloDF13 ori, lacI, double PT7lac, StrepR; pCDFduet-1 carrying TAL | Rodrigues et al., 2015a |
| pETduet_TAL | ColE1(pBR322) ori, lacI, double PT7lac, AmpR; pETduet-1 carrying TAL | Rodrigues et al., 2015a |
| pETduet_TAL_C3H | pETduet_TAL carrying codon-optimized C3H from Saccharothrix espanaensis | Rodrigues et al., 2015a |
| pRSFduet_C3H | pRSFduet-1 carrying C3H | Rodrigues et al., 2015a |
| pCDFduet_C3H | CloDF13 ori, lacI, double PT7lac, StrepR; pCDFduet-1 carrying C3H | Rodrigues et al., 2015a |
| pCDFduet_DCS | CloDF13 ori, lacI, double PT7lac, StrepR; pCDFduet-1 carrying codon-optimized DCS from Curcuma longa | Rodrigues et al., 2015b |
| pRSFduet_CURS1 | pRSFduet-1 carrying codon-optimized CURS1 from C. longa | Rodrigues et al., 2015b |
| pRSFduet_CURS1_CCoAOMT | pRSFduet_CURS1 carrying codon-optimized CCoAOMT from Medicago sativa | Rodrigues et al., 2015b |
| pAC-4CL1 | p15A ori, Plac, CmR, pACYC184-derived plasmid carrying 4CL1 from A. thaliana | Addgene (35947) |
| pACYCduet_COMT | pACYCDuet-1 carrying COMT | This study |
| pRSFduet_COMT | pRSFduet-1 carrying COMT | This study |
| pAC_COMT | pAC-4CL1 without 4CL1 and carrying COMT | This study |
| pACYCduet_4CL1 | pACYCduet-1 carrying 4CL1 | This study |
| pRSFduet_CURS1_COMT | pRSFduet_CURS1 carrying COMT | This study |
| pCDFduet_DCS_C3H | pCDFduet_DCS carrying C3H | This study |
| pACYCduet_4CL1_TAL | pACYCduet_4CL1 carrying TAL | This study |
| pRSFduet_COMT_C3H | pRSFduet_COMT carrying C3H | This study |
| pRSFduet_CURS1_4CL1 | pRSFduet_CURS1 carrying COMT | This study |
| pRSFduet_CURS1_DCS | pRSFduet_CURS1 carrying DCS | This study |
| pRSFduet_TAL_C3H | pRSFduet_TAL carrying C3H | This study |