Table 2.
Escherichia coli strains and plasmids used in this study
| Strain/plasmid | Description | Source |
|---|---|---|
| Strains | ||
| DH5ɑ | supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Invitrogen |
| DH5α λpir | supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17 LAMpir U169 recA1 endA1 gyrA96 thi-1 relA1 | Lab stock |
| W3110 | Wide type, λ-F-mcrA mcrB IN (rrnD-rrnE)1 | DSM5911 |
| EC101 | W3110 with plasmid pET-trc-IEB | This study |
| EC102 | W3110 with plasmid pCDF-yciG-IEB | This study |
| EC103 | W3110 with plasmid pCDF-pstA-IEB | This study |
| EC104 | W3110 with plasmid pCDF-yodA-IEB | This study |
| EC105 | W3110 with plasmid pCDF-astC-IEB | This study |
| EC106 | W3110 with plasmid pCDF-ybiM-IEB | This study |
| EC201 | W3110 with plasmid pACYC-phnI | This study |
| EC202 | W3110 with plasmid pACYC-phoR | This study |
| EC203 | W3110 with plasmid pACYC-phnF | This study |
| EC204 | W3110 with plasmid pACYC-phnC | This study |
| EC205 | W3110 with plasmid pACYC-phnD | This study |
| EC301 | W3110 with plasmid pACYC-yfiL | This study |
| EC302 | W3110 with plasmid pACYC-yijF | This study |
| EC303 | W3110 with plasmid pACYC-cysP | This study |
| EC304 | W3110 with plasmid pACYC-yejG | This study |
| EC305 | W3110 with plasmid pACYC-yhcN | This study |
| EC401 | DH5α λpir with plasmid pRC-IS5 | This study |
| EC-IS5 | W3110 with the integration of plasmid pRC-IS5 | This study |
| EC-IS5(ΔCm) | EC-IS5 with the deletion of Cm | This study |
| Plasmidsa | ||
| pCDFDuet-1 | pCloDF13-derived vector; T7 promoter, StrR | Lab stock |
| pET-30a-trc | pBR322-derived vector; trc promoter, KmrR | Lab stock |
| pACYCDuet-1 | p15A-derived vector; T7 promoter, CmrR | Lab stock |
| pTrc99a | pBR322-derived vector; trc promoter, AmpR | [26] |
| pRC43 | Including R6K ori, Cm, IS5 sequence | [29] |
| pCas | repA101(Ts) kan Pcas-cas9 ParaB-Red lacIq Ptrc-sgRNA-pMB1 | [14] |
| pTargetF-cadA | pMB1 aadA sgRNA-cadA | [14] |
| pET-trc-IEB | Lycopene synthetic module under the trc promoter | This study |
| pCDF-yciG-IEB | Lycopene synthetic module under the yciG promoter | This study |
| pCDF-pstA-IEB | Lycopene synthetic module under the pstA promoter | This study |
| pCDF-yodA-IEB | Lycopene synthetic module under the yodA promoter | This study |
| pCDF-astC-IEB | Lycopene synthetic module under the astC promoter | This study |
| pCDF-ybiM-IEB | Lycopene synthetic module under the ybiM promoter | This study |
| pACYC-phnI | Lycopene synthetic module under the yciG promoter, feedforward module under the phnI promoter | This study |
| pACYC-phoR | Lycopene synthetic module under the yciG promoter, feedforward module under the phoR promoter | This study |
| pACYC-phnF | Lycopene synthetic module under the yciG promoter, feedforward module under the phnF promoter | This study |
| pACYC-phnC | Lycopene synthetic module under the yciG promoter, feedforward module under the phnC promoter | This study |
| pACYC-phnD | Lycopene synthetic module under the yciG promoter, feedforward module under the phnD promoter | This study |
| pACYC-yfiL | Lycopene synthetic module under the yciG promoter, feedback module under the yfiL promoter | This study |
| pACYC-yijF | Lycopene synthetic module under the yciG promoter, feedback module under the yijF promoter | This study |
| pACYC-cysP | Lycopene synthetic module under the yciG promoter, feedback module under the cysP promoter | This study |
| pACYC-yejG | Lycopene synthetic module under the yciG promoter, feedback module under the yejG promoter | This study |
| pACYC-yhcN | Lycopene synthetic module under the yciG promoter, feedback module under the yhcN promoter | This study |
| pTargetF-delete | gRNA for N20PAM | This study |
| pRC-IS5 | Including R6K ori, Cm, fragmentary IS5 sequence, Lycopene synthetic module under the trc promoter, feedforward module under the phoR promoter and feedback module under the yejG promoter | This study |
a AmpR: ampicillin; KanR: kanamycin; StrR: Streptomycin; Cm: chloramphenicol; R:resistance