Table 1 Strains and plasmids used in this study
Strains or plasmids | Relevant characteristics | Source |
|---|---|---|
Strains | ||
 E. coli DH5α | F-φ80 lacZΔM15Δ (lacZYA-argF) U169recA1 endA1 hsdR17 (rk -, mk +) phoA, sup E44 thi − 1 gyr A96 rel A1 λ - | TransGen Biotech |
 E. coli Top10 | F-mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15ΔlacX74recA1 araΔ139Δ(ara-leu)7697galU galK rpsL(Sm) endA1nupG | TransGen Biotech |
 P. putida KT2440 | Wild-type strain | In our lab |
 Pp1 | P. putida KT2440 + pBBR1MCS2 | This study |
 Pp2 | P. putida KT2440 + pBBR-Cas9 | This study |
 Pp3 | P. putida KT2440 + pBBR-Cas9ΔyqhD-HA500 | This study |
 PPΔyqhD | P. putida KT2440 with yqhD deletion | This study |
 PPΔevf | P. putida KT2440 with ech, vdh and fcs deletions | This study |
 PpΔend | P. putida KT2440 with endAX deletions | This study |
 PpV01 | PpΔend + pBBR-vs. | This study |
 PpV02 | PpΔend + pBBR-vs-dxs | This study |
 PpV03 | PpΔend + pBBR-vs-ispA | This study |
 PpV04 | PpΔend with phaG deleted + pBBR-vs-dxs | This study |
 PpV05 | PpΔend with phaG and phaC1ZC2 deleted + pBBR-vs-dxs | This study |
 PpV06 | PpΔend with phaG and phaC1ZC2 deleted + pBBR-vs-dxs-ispA | This study |
 PpV07 | PpΔend with phaG and phaC1ZC2 deleted and vs. linked to ispA in genome + pBBR-vs-dxs-ispA | This study |
Plasmids | Â | Â |
 pBBR1MCS2 | p1, pBBR replicon, mob+, Km | In our lab |
 pBBR-Cas9 | p2, pBBR1MCS2 derivative, Plac and Cas9 | This study |
 pBBR-Cas9ΔyqhD-HA500 | p3, pBBR-Cas9 derivative, sgRNA and donor DNA (500 bp per arm) of yqhD | This study |
 pBBR-Cas9ΔyqhD-HA300 | pBBR-Cas9 derivative, sgRNA and donor DNA (300 bp per arm) of yqhD | This study |
 pBBR-Cas9ΔyqhD-HA150 | pBBR-Cas9 derivative, sgRNA and donor DNA (150 bp per arm) of yqhD | This study |
 pBBR-Cas9ΔyqhD-HA900 | pBBR-Cas9 derivative, sgRNA and donor DNA (900 bp per arm) of yqhD | This study |
 pBBR-Cas9ΔyqhD-HA1200 | pBBR-Cas9 derivative, sgRNA and donor DNA (1200 bp per arm) of yqhD | This study |
 pBBR-Cas9Δevf-HA150 | pBBR-Cas9 derivative, sgRNA and donor DNA (150 bp per arm) of ech-vdh-fcs | This study |
 pBBR-Cas9Δevf-HA300 | pBBR-Cas9 derivative, sgRNA and donor DNA (300 bp per arm) of ech-vdh-fcs | This study |
 pBBR-Cas9Δevf-HA500 | pBBR-Cas9 derivative, sgRNA and donor DNA (500 bp per arm) of ech-vdh-fcs | This study |
 pBBR-Cas9Δevf-HA900 | pBBR-Cas9 derivative, sgRNA and donor DNA (900 bp per arm) of ech-vdh-fcs | This study |
 pBBR-Cas9Δevf-HA1200 | pBBR-Cas9 derivative, sgRNA and donor DNA (1200 bp per arm) of ech-vdh-fcs | This study |
 pBBR-Cas9ΔcatAB | pBBR-Cas9 derivative, sgRNA and donor DNA of catAB deletion | This study |
 pBBR-Cas9Δvdh | pBBR-Cas9 derivative, sgRNA and donor DNA of vdh deletion | This study |
 pBBR-Cas9ΔvanAB | pBBR-Cas9 derivative, sgRNA and donor DNA of vanAB deletion | This study |
 pBBR-Cas9ΔyqhD::gfp | pBBR-Cas9 derivative, sgRNA and donor DNA for gfp replacing yqhD | This study |
 pBBR-Cas9ΔyqhD::rfp | pBBR-Cas9 derivative, sgRNA and donor DNA for rfp replacing yqhD | This study |
 pBBR-Cas9Δ2827::MluI | pBBR-Cas9 derivative, sgRNA and donor DNA for MluI replacing PP_2827 | This study |
 pBBR-Cas9::RBS-fadH | pBBR-Cas9 derivative, sgRNA and donor DNA for an RBS insertion before fadH | This study |
 pBBR-Cas9ΔendA | pBBR-Cas9 derivative, sgRNA and donor DNA of endA deletion | This study |
 pBBR-Cas9ΔendX | pBBR-Cas9 derivative, sgRNA and donor DNA of endX deletion | This study |
 pBBR-Cas9ΔphaG | pBBR-Cas9 derivative, sgRNA and donor DNA of phaG deletion | This study |
 pBBR-Cas9ΔphaC1ZC2 | pBBR-Cas9 derivative, sgRNA and donor DNA of phaC1ZC2 deletion | This study |
 pBBR-Cas9::vs. | pBBR-Cas9 derivative, sgRNA and donor DNA for vs. insertion linking with ispA | This study |
 pBBR-vs. | pBBR1MCS2 derivative, carrying vs. (valencene synthase) gene | This study |
 pBBR-vs-dxs | pBBR1MCS2 derivative, carrying vs. and dxs (DXP synthase) genes | This study |
 pBBR-vs-ispA | pBBR1MCS2 derivative, carrying vs. and ispA (GPP/FPP synthase) genes | This study |
 pBBR-vs-dxs-ispA | pBBR1MCS2 derivative, carrying vs., dxs and ispA | This study |