TABLE 3.
Strains and plasmids used in this study
| Strain(s) or plasmid | Characteristics | Reference or source |
|---|---|---|
| Strains | ||
| P. putida S12 | Wild-type P. putida S12 (ATCC 700801), harboring megaplasmid pTTS12, solvent-tolerant strain | 54 |
| P. putida S12d (S12-06, S12-10, S12-22) | P. putida S12 ΔpTTS12, non-solvent-tolerant strains | 19 |
| P. putida S12e (S12-06e30, S12-10e35, S12-22e30) | ALE-derived P. putida S12 ΔpTTS12, solvent-tolerant strain | This paper |
| P. putida S12e30 | ALE-derived wild-type P. putida S12 (control strain) | This paper |
| P. putida S12d ΔarpR | P. putida S12 ΔpTTS12 ΔRPPX_14650 | This paper |
| P. putida S12d Δafr | P. putida S12 ΔpTTS12 ΔRPPX_14685 | This paper |
| P. putida S12d ΔgacA | P. putida S12 ΔpTTS12 ΔRPPX_00635 | This paper |
| P. putida S12d ΔgacS | P. putida S12 ΔpTTS12 ΔRPPX_15700 | This paper |
| P. putida S12e arpR-wt | ALE-derived P. putida S12 ΔpTTS12 RPPX_14650-wt | This paper |
| P. putida S12e afr-wt | ALE-derived P. putida S12 ΔpTTS12 RPPX_14685-wt | This paper |
| P. putida S12e gacS-wt | ALE-derived P. putida S12 ΔpTTS12 RPPX_15700-wt | This paper |
| P. putida S12e rpoB′-wt | ALE-derived P. putida S12 ΔpTTS12 RPPX_06985-wt | This paper |
| P. putida S12e ATP-wt | ALE-derived P. putida S12 ΔpTTS12 RPPX_09480-09510-wt | This paper |
| P. putida S12 ΔlapA | P. putida S12 ΔRPPX_08475 | This paper |
| P. putida S12 Δflg | P. putida S12 ΔRPPX_02040-02125 | This paper |
| P. putida S12-RE1 | P. putida S12-10 (ΔpTTS12) ΔarpR | This paper |
| P. putida S12-RE2 | P. putida S12-10 (ΔpTTS12) ΔarpR ΔgacS | This paper |
| P. putida S12-RE3 | P. putida S12-10 (ΔpTTS12) ΔarpR ΔgacS atpα (R165C) | This paper |
| P. putida S12-RE4 | P. putida S12-10 (ΔpTTS12) ΔarpR ΔgacS atpα (R165C) Δafr | This paper |
| P. putida S12-RE5 | P. putida S12-10 (ΔpTTS12) ΔarpR Δgac; atpα (R165C) Δafr rpoB′ (D622G) | This paper |
| E. coli WM3064 | thrB1004 pro thi rpsL hsdS lacZΔM15 RP4-1360 Δ(araBAD)567 ΔdapA1341::[erm pir] | William Metcalf |
| Plasmids | ||
| pEMG | Kmr Apr ori R6K lacZα MCS flanked by two I-SceI sites | 49 |
| pEMG-ΔarpR | pEMG plasmid for constructing P. putida S12d Δafr | This paper |
| pEMG-Δafr | pEMG plasmid for constructing P. putida S12d Δafr | This paper |
| pEMG-ΔgacA | pEMG plasmid for constructing P. putida S12d ΔgacA | This paper |
| pEMG-ΔgacS | pEMG plasmid for constructing P. putida S12d ΔgacS | This paper |
| pEMG-ΔlapA | pEMG plasmid for constructing P. putida S12 ΔlapA | This paper |
| pEMG-Δflg | pEMG plasmid for constructing P. putida S12 Δflg | This paper |
| pEMG-c-arpR | pEMG plasmid for constructing P. putida S12e arpR-wt | This paper |
| pEMG-c-afr | pEMG plasmid for constructing P. putida S12e afr-wt | This paper |
| pEMG-c-gacS | pEMG plasmid for constructing P. putida S12e gacS-wt | This paper |
| pEMG-c-rpoB′ | pEMG plasmid for constructing P. putida S12e rpoB′-wt | This paper |
| pEMG-c-ATP | pEMG plasmid for constructing P. putida S12e ATP-wt | This paper |
| pSW-2 | Gmr ori RK2 xylS Pm → I-SceI | 49 |
| p421-cas9 | Cas9 and tracrRNA; oriV RK2; Smr/Spr | 51 |
| p658-ssr | xylS ‐Pm → ssr oriV RSF1010; Gmr | 51 |
| p2316 | SEVA CRISPR array; oriV pBBR1; Kmr | 50 |
| p2316-ATPsyn | pSEVA2316 derivative containing the ATP synthase spacer | This paper |
| p2316-rpoB′-622 | pSEVA2316 derivative containing the rpoB′ spacer (substitution of amino acid D622G) | This paper |