Table 1 List of E. coli strains and plasmids used in this study
Strains and plasmids | Description | Source or accession number |
|---|---|---|
Strains | ||
W3110 | F-, λ−, IN(rrnD-rrnE)1, rph-1 | CGSC #4474 |
BL21 | F– ompT gal dcm lon hsdSB(rB–mB–) [malB+]K-12(λS) | NEB #C2530H |
BL21(DE3) | F– ompT gal dcm lon hsdSB(rB–mB–) λ(DE3 [lacI lacUV5-T7p07 ind1 sam7 nin5]) [malB+]K-12(λS) | NEB #C2527I |
W3110::IL5 | F-, λ−, IN(rrnD-rrnE)1, rph-1 attB (HK022)::lacI- PLacUV5- lacZ’- T7 gene 1 | This study |
W3110::L5 | F-, λ−, IN(rrnD-rrnE)1, rph-1 attB (HK022):: PLacUV5- lacZ’- T7 gene 1 | This study |
W3110::pI | F-, λ−, IN(rrnD-rrnE)1, rph-1 attB (HK022):: PLacI- T7 gene 1 | This study |
Plasmids | ||
pAH69 | 4951 bp, AmpR, Rep101 ori, repA101(ts), HK022 integrase, cIts857 protein | (Haldimann and Wanner 2001) |
pHK-Km | 2539 bp, KanR, R6K ori, FRT- HK022 attp- FRT | (Chen et al. 2011) |
pDS3.0- HsdNC- T7RNAP | 11,966 bp, AmpR, GmR, R6K ori, Hsd-N, PLacI promoter, T7 gene1, Hsd-R, sacB | (Yi and Ng 2020) |
pHK-IL5-Km | 6933 bp, KanR, R6K ori, FRT- HK022 attp- lacI-PLacuv5- T7 promoter- lacZ’- T7 gene1- FRT | This study |
pHK-L5-Km | 5766 bp, KanR, R6K ori, FRT- HK022 attp- PLacuv5- T7 promoter- lacZ’- T7 gene1- FRT | This study |
pHK-pI-Km | 6586 bp, KanR, R6K ori, FRT- HK022 attp- Hsd-N- PLacI promoter- T7 gene1- Hsd-R- FRT | This study |
pET28a (+)-sfGFP | 6013 bp, KanR, pBR322 ori, lacI, T7 promoter, lacO, sfGFP | This study |
pSU-T7-sfGFP | 9471 bp, CmR, pUC ori, T7 promoter, sfGFP | (Tan and Ng 2020) |
pSU-PlacI-sfGFP | 3099 bp, CmR, pUC ori, PlacI promoter, sfGFP | (Yi and Ng 2020) |
pSU-PlacUV5-sfGFP | 3099 bp, CmR, pUC ori, PlacUV5 promoter, sfGFP | This study |
pSU- PlacUV5-NRBS-sfGFP | 3099 bp, CmR, pUC ori, PlacUV5 promoter, native RBS, sfGFP | This study |
pSU-PlacUV5-LacZα’-sfGFP | 3571 bp, CmR, pUC ori, PlacUV5 promoter, LacZα’, sfGFP | This study |
pET32a-SyCA | 6595 bp, AmpR, pBR322 ori, trxA, lacI, T7 promoter, lacO, SyCA | (Hsu et al. 2019) |
pET21a (+)-Cas9 | 9471 bp, AmpR, pBR322 ori, lacI, T7 promoter, lacO, SpCas9-6His | Lab stock |
pET20b (+)-Cas9 | 7694 bp, AmpR, pBR322 ori, ROP, T7 promoter, Spcas9-6 His | This study |
pET28a (+)-CadA | 7498 bp, KanR, pBR322 ori, lacI, T7 promoter, lacO, cadA | This study |
pSU-T7-CadA | 4255 bp, CmR, pUC ori, T7 promoter, cadA | This study |