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Table 1 Strains and plasmids used in this study

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Unpublished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Unpublished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