Table 2.
PCR primers and synthesised gene fragments used in the construction of plasmids for this study. Underlined sequences indicate restriction sites. Italic sequences indicate overlap regions for Gibson assembly. Complete sequences for each plasmid, including details of the primer binding sites, promoters, ribosome binding sites and replication origins are available online from Addgene under accession numbers 128848 – 128853 and 128855 – 128857.
| Name | Sequence |
|---|---|
| PCR primers | |
| K12-N_5p_Eco_F | ATATATGAATTCCTGAATTGCGCAAAGTTTACG |
| K12-N_5p_Bam_R | ATATATGGATCCATTGCAAGTCGTTGATTACGT |
| K12-N_3p_Xho_F | ATATATCTCGAGGATTAACTGAGACTGACGG |
| K12_3p_Nhe_R2 | ATATATGCTAGCTAAATTCCAGGCAGAAAAAAAC |
| RK2ori_Nhe_F | ATATATGCTAGCGCAGAGCCATGTAGGG |
| RK2ori_Hin_R | ATATATAAGCTTCCAGGCATCAAATAAAACGA |
| 372F | CGATCGCCGTTTTCGTATCG |
| CC1 | CTTAAGTTCTACGTGTTCCGC |
| 2836R | TGTAGGGCGTCATAGCGTTC |
| CC2 | CATGCTGGAGTTCTTCGCC |
| idnT_F | CTGTTTAGCGAAGAGGAGATGC |
| idnT_R | ACAAACGGCGGCGATAGC |
| hcaT_F | GCTGCTCGGCTTTCTCATCC |
| hcaT_R | CCAACCACGCAGACCAACC |
| cysG_F | TTGTCGGCGGTGGTGATGTC |
| cysG_R | ATGCGGTGAACTGTGGAATAAACG |
| fliC192_F | GGCCCGTAACGCCAACGACG |
| fliC289_R | CCGTCAGTTCACGCACACGC |
| 1F | CTACATGGCTCTGCGCTAGCGCTGAGGTCCCGCAGC |
| 4573R | TATTTGATGCCTGGAAGCTTCACTGCCCGCTTTCCAGT |
| G_Nhe_F | GCTAGCGCAGAGCCATGTAGGG |
| G_Hin_R | AAGCTTCCAGGCATCAAATAAAACGA |
| Synthesised gene fragments | |
| rhaBAD promoter + RBS | ATATATGTCGACCACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAATGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGTATTCAGGCGCTTTTTAGACTGGTCGTAATGAAATTCAGCAGGATCACACCATGGATATAT |
| pUC origin | ATATATAAGCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAGCTAGCATATAT |