Table 1. Bacterial strains, plasmids and oligonucleotides used in this work.
Underlined bases indicate oligonucleotide positions that differ from the corresponding templates.
| Bacterial strain | Genotype/phenotype | Reference/source |
| E. coli | ||
| DH5α | Ψ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rk- mk+) supE44 thi-1 gyrA relA1 | [71] |
| P. putida | ||
| KT2440 | mt-2 hsdR1 (r- m+) | [72] |
| KT2442 | mt-2 hsdR1 (r- m+) Rifr | [72] |
| KT2440 fliA::aphA-3 | KT2440 fliA::Km. Kmr | [46] |
| MRB49 | KT2442 flhF::miniTn5-Km. Rifr Kmr | [45] |
| MRB52 | KT2442 ΔfleQ | This work |
| MRB69 | KT2442 ΔflhF | This work |
| MRB71 | KT2442 ΔfleN | This work |
| MRB78 | KT2442 ΔflhF-fleN | This work |
| MRB101 | KT2442 ΔfleN ΔfleQ | This work |
| Plasmid | Genotype/phenotype | Reference/source |
| pENTR/D-TOPO | Vector for directional TOPO cloning. Kmr | Thermo Fisher Scientific |
| pEX18-Tc | Gene replacement vector with MCS from pUC18. Tcr Sacs | [73] |
| pMPO284 | pPS854-derived vector containing pUTminiTn5Km Kmr gene flanked by the FRT sites, Apr Kmr | [26] |
| pMRB1 | pBBR1-MCS4-derived broad host-range gfpmut3::lacZ transcriptional fusion vector. Apr | [26] |
| pMRB2 | pBBR1-MCS4-derived vector containing the Gateway conversion cassette attR1-Cmr-ccdB-attR2. Apr Cmr | [26] |
| pMRB3 | pBBR1-MCS4-derived vector containing the Gateway conversion cassette attR1-Cmr-ccdB-attR2. Apr Cmr | [23] |
| pMRB67 | pMRB2-derived vector containing a PlapA-gfpmut3-lacZ transcriptional fusion. Apr | [23] |
| pMRB89 | Expression vector for E. coli YhjH containing nahR-Psal-TnasF-yhjH. Strr | [26] |
| pMRB95 | pTYB12-based vector for FleQ overproduction. Apr | This work |
| pMRB96 | pEX18-Tc with Kmr gene flanked by FRT sites cloned between fleQ flanking regions. Tcr Kmr Sacs | This work |
| pMRB104 | pEX18-Tc with Kmr gene flanked by FRT sites cloned between flhF flanking regions. Tcr Kmr Sacs | This work |
| pMRB105 | pEX18-Tc with Kmr gene flanked by FRT sites cloned between fleN flanking regions. Tcr Kmr Sacs | This work |
| pMRB112 | pMRB2-derived vector containing a PbcsD-gfpmut3-lacZ transcriptional fusion. Apr | [23] |
| pMRB115 | pMRB3-derived vector containing a PflhA-gfpmut3-lacZ transcriptional fusion. Apr | [23] |
| pMRB132 | pTYB12-based vector for FleN overproduction. Apr | This work |
| pMRB144 | pEX18-Tc with Kmr gene flanked by FRT sites cloned between flhF-fleN flanking regions. Tcr Kmr Sacs | This work |
| pMRB158 | pMRB3-derived vector containing a PflhF-gfpmut3-lacZ transcriptional fusion. Apr | This work |
| pRK2013 | Helper plasmid for triparental mating. ColE1 replicon. Kmr | [74] |
| pTYB12 | Expression vector for N-terminal Intein–chitin-binding domain protein fusions. Apr | New England Biolabs |
| pYedQ | pRK404A-derived plasmid expressing E. coli YedQ from the Plac promoter. Tcr | [75]. |
| Oligonucleotide | Sequence (5’ to 3’) | |
| flhF-up-Fwd | ACCCGAATTCCGAACTGGCGATCAACCC | |
| flhF-up-Rev | CGCAGGATCCGCATTATCCCCTACCTCA | |
| flhF-down-Fwd | CGGCGGATCCGTTGACCATGAAGCGTGT | |
| flhF-down-Rev | TGCAAAGCTTGCTCGACAAAGAACTCCA | |
| fleN-up-Fwd | CCTGGAATTCCTGCTGGAAGTGCAACTC | |
| fleN-up-Rev | TGCAGGATCCCCATGTCTGTTCTTTACC | |
| fleN-down-Fwd | GGACGGATCCTATGAACGCCAGCGGCTT | |
| fleN-down-Rev: | TTTTAAGCTTCTTGTCCAATTAGACCTC | |
| flhA-RT-fwd | TCTTCACGTTCAACATCGCC | |
| flhA-RT-rev | TCCATCGAGCCGTAGAACTC | |
| Pint_FlhA_Fwd | CACCCTCTGAAGACATGGGCAAG | |
| Pint_FlhA_Rev2 | CACGGACCAGTTTCATGGCC | |
| flhA-flhF-RT-fwd | CTTCTGGAACCTAGCATGGC | |
| flhA-flhF-RT-rev | AATGCGCGTGTGGGTCTT | |
| flhF-miniTn5DST2-fwd | CATGCAACTGGAAACCTTGG | |
| flhF-miniTn5DST2-rev | GCCGTGGGTTGTGATAGAGA | |
| flhF-fleN-RT-fwd | GAGCCTTGCCATCAGTCATG | |
| flhF-fleN-RT-rev | CAGCAACACGTCGACATTGG | |
| fleN-fliA-RT-fwd | GACCGCTTCCTTGACGTTG | |
| fleN-fliA-RT-rev | CGTCGTATTTGTTGGCCACT | |
| fliA-cheY-RT-fwd | CAAGGAAATCGGTGAGGTGC | |
| fliA-cheY-RT-rev | CCGCTTCAATGATCTGGTCG | |
| PflhF-fwd | CACCCTCTGAAGACATGGGCAAG | |
| PflhF-rev | CACGGACCAGTTTCATGGCC | |
| fleN-fwd-NdeI | AACACATATGGGTAGCATGCATCCC | |
| fleN-rev-EcoRI | CCGCGAATTCTCATAGTACGGGTCCCGC | |
| PlapA1-fwd-HindIII | TGACAAGCTTCGACTGACTTCGGATTCC | |
| PlapA1-rev | AGCTATGTGTCGCAAATCAA | |
| PlapA2-fwd | TTGATTTGCGACACATAGCT | |
| PlapA2-rev-BamHI | GCCCGGATCCATTGGACTCTCCGTGTGACC | |
| PbcsD-fwd-HindIII | CCCAAGCTTGTTGATCGCCAGCACCTGG | |
| PbcsD-rev | GACTCATGTCAAAAAAACGACAAAAATGA | |
| lapA-qRT-fwd | AGCATTGTCGGCCAGGTTATT | |
| lapA-qRT-rev | TCGATAAGTACACGCCGGATG | |
| bcsD-qRT-fwd | CATTCCGCCTTCAAACCGTTT | |
| bcsD-qRT-rev | GGAAATCGCCAACAACTGCAT |