From: CRISPRi-mediated metabolic engineering of E. coli for O-methylated anthocyanin production
Strain or plasmid name | Properties/genotype | References |
---|---|---|
E. coli strains | ||
 E. coli DH5α™ | F-Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK−, mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | Novagen |
 E. coli BL21 Star™ (DE3) | F-ompT hsdS B (rB−mB−) gal dcm rne131 (DE3) | Invitrogen |
E. coli plasmids | ||
 pMAL-c2X-PhANS | ColE1(AmpR), Petunia hybrida anthocyanidin synthase (PhANS) with N-terminal cytosolic MBP tag | Unpublished |
 pMAL-c2X-At3GT | ColE1(AmpR), Arabidopsis thaliana 3-O-glucosyltransferase (At3GT) with N-terminal cytosolic MBP tag | Unpublished |
 pETM6-PhANS | ColE1(AmpR), ePathBrick feature, PhANS | This study |
 pETM6-At3GT | ColE1(AmpR), ePathBrick feature, At3GT | This study |
 pETM6-MBP-PhANS | ColE1(AmpR), ePathBrick feature, PhANS with N-terminal MBP tag | This study |
 pETM6-MBP-At3GT | ColE1(AmpR), ePathBrick feature, At3GT with N-terminal MBP tag | This study |
 pETM6-At3GT-m-PhANS | ColE1(AmpR), ePathBrick feature, PhANS and At3GT in monocistronic configuration | This study |
 pACM4-At3GT-m-PhANS | pACYC184(CmR), ePathBrick feature, PhANS and At3GT in monocistronic configuration | This study |
 pETM6-MBP-At3GT-m-MBP-PhANS | ColE1(AmpR), ePathBrick feature, PhANS and At3GT with N-terminal MBP tags in monocistronic configuration | This study |
 pETM6-VvAOMT1 | ColE1(AmpR), ePathBrick feature, Vitis vinifera anthocyanin O-methyltransferase (VvAOMT1) | This study |
 pETM6-CkmOMT2 | ColE1(AmpR), ePathBrick feature, fragrant cyclamen ‘Kaori-no-mai’ anthocyanin O-methyltransferase 2 (CkmOMT2) | This study |
 pETM6-PhMF1 | ColE1(AmpR), ePathBrick feature, Petunia hybrida anthocyanin O-methyltransferase 1 (PhMF1) | This study |
 pETM6-PhMF2 | ColE1(AmpR), ePathBrick feature, Petunia hybrida anthocyanin O-methyltransferase 2 (PhMF2) | This study |
 pETM6-SlAOMT | ColE1(AmpR), ePathBrick feature, Solanum lycopersicum anthocyanin O-methyltransferase (SlAOMT) | This study |
 pETM6-VvAOMT1-m-At3GT-m-PhANS | ColE1(AmpR), ePathBrick feature, VvAOMT1, PhANS, and At3GT in monocistronic configuration | This study |
 pETM6-VvAOMT1-m-MBP-At3GT-m-MBP-PhANS | ColE1(AmpR), ePathBrick feature, VvAOMT1 and N-terminal MBP-tagged PhANS and At3GT in monocistronic configuration | This study |
 pETM6-CkmOMT2-m-At3GT-m-PhANS | ColE1(AmpR), ePathBrick feature, CkmOMT2, PhANS, and At3GT in monocistronic configuration | This study |
 pETM6-CkmOMT2-m-MBP-At3GT-m-MBP-PhANS | ColE1(AmpR), ePathBrick feature, CkmOMT2 and N-terminal MBP-tagged PhANS and At3GT in monocistronic configuration | This study |
 pdCas9 | pACYC184(CmR), tracrRNA, cas9(D10A, H840A), non-targeting CRISPR spacer with two BsaI sites | [49] |
 pdCas9-M-MetJ1 | pACYC184(CmR), tracrRNA, cas9(D10A, H840A), CRISPR spacer targeting MetJ promoter at position 1 | This study |
 pdCas9-M-MetJ2 | pACYC184(CmR), tracrRNA, cas9(D10A, H840A), CRISPR spacer targeting MetJ promoter at position 2 | This study |