Table 1.
Oligonucleotides used in this work
| Primer | Sequence (5′ to 3′)a |
|---|---|
| PtcdR-SNAP-EcoRI-Fw | CCCTTAAGCGAATTCTAAAAGTAAACG |
| PtcdR-SNAP-XhoI-Rev | CATAAAATCCTCGAGTCTTATATTTATAATG |
| PtcdA-SNAP-EcoRI-Fw | CACAAAGATGAATTCTGGTCAGTTGGT |
| PtcdA-SNAP-XhoI-Rev | GTATTATTACTCGAGATAATAAATCCAC |
| PtcdB-SNAP-EcoRI-Fw | TATCAAAGTGAATTCGTTTTTGAGGAAG |
| PtcdB-SNAP-XhoI-Rev | CTATAATACTCGAGCATCTAAATGCTAAAAC |
| PtcdC-SNAP-EcoRI-Fw | ACTTCACCTGAATTCTGGTATATTC |
| PtcdC-SNAP-XhoI-Rev | CATAATACAATCCTCGAGTTATTAGAT |
| PtcdE-SNAP-EcoRI-Fw | CAATTGGAATTCGATGGATATATGATATG |
| PtcdE-SNAP-XhoI-Rev | CATTCATCATAGCTCGAGTTTTTATTG |
| PtcdRA-SNAP-EcoRI-Fw | CCCCCCGAATTCTACTTTATTTATTAGAAAAA |
| PsigDmut-Fw | GCATATTTTCATATAAAATTTAATTTATTTGCATCTTCTATAATTATAATG |
| PsigDmut-Rev | CATTATAATTATAGAAGATGCAAATAAATTAAATTTTATATGAAAATATGC |
| TcdR-RBSopt-Fw | GCGCGCCTCGAGGGAGGAACTACTATGCAAAAGTCTTTTTATG |
|
TcdR-comp-HindIII-Rev PsspA-BamHI-Fw |
CCCAAGCTTATTAATTTGCTCTTC AGATGAGGAGGATCCGGATAAAAGAGTTC |
| PsspA-SNAP-XhoI-Rev | CTTCCTTCTCTCGAGTTTATTTTGTGTTTGC |
| PspoIIIAA-BamHI-Fw | TAGATGGTGGGATCCCTAGGGCTTACCAAAAAAC |
| PspoIIIAA-SNAP-XhoI-Rev | GTTTATTCATCTCTTGCTCGAGTCCTTG |
| TcdR-comp-XhoI-Rev | CGCGCTCGAGATTAATTTGCTCTTC |
| TcdR-NcoI-Fw | CCCCCATGGAAAAGTCTTTTTATG |
| TcdR-SalI-Rev | CCCGTCGACCAAGTTAAAATAATTTTC |
| CDSigGpET28a-Fw | TGCCTCGAGTACATATTTTCTCATATTTTTTAAAGC |
| CDSigGpET28a-Rev | AGGGGGTGACCCCATGGCAGCTCTTAAATC |
| SigD-NcoI-Fw | CCCCCATGGATAGAGAAGAATTAATAAAAG |
| SigD-XhoI-Rev | CCCCTCGAGTATAGAATATTTAAGTTC |
| SpoVT-NdeI-Fw | CCCCATATGAAAGCAACAGGTATAGTTAG |
| SpoVT-XhoI-Rev | CCCCTCGAGTTATTGAACTTGTTTTCC |
| tcdR-AscI-Fw | CCCCGGCGCGCCATTATCTTAAGAGAGGAG |
| tcdR-SOE-Rev | CATAAATAAAATTTCTTGCAAATCATC |
| tcdR-SOE-Fw | TTGCAAGAAATTTTATTTATGGAAAATTATTTTAACTTG |
| tcdR-SbfI-Rev | CCCCCCTGCAGGTATCTATATAAATATCTG |
| tcdR-vef-Fw | GTATCATTTCACGAAGAGG |
| tcdR-vef-Rev | GGGTCATTTAAGTTTTCTC |
| tcdR-comp-BamHI-Fw | CCCGGATCCTAAAAATATTTTGATATG |
| tcdR-comp-HindIII-Rev | CCCAAGCTTATTAATTTGCTCTTC |
| tcdA-AscI-Fw | CCCCGGCGCGCCGGTAGTATATCAAACATTGG |
| tcdA-SOE-Rev | CTCATTTTCTCTTGGTCTAATGCTATATGCGAG |
| tcdA-SOE-Fw | CCAAGAGAAAATGAGCCTGGGATATATGGC |
| tcdA-SbfI-Rev | CCCCCCTGCAGGGATAAGGTTGTACTATGTAG |
| tcdB-AscI-Fw | CCCCGGCGCGCCCAAAGTAAGTCTGTTTTTGAGG |
| tcdB-SOE-Rev | CAATATTGCAACATATTCATCTTCTTG |
| tcdB-SOE-Fw | GAATATGTTGCAATATTGCAATTAGTG |
| tcdB-SbfI-Rev | CCCCCCTGCAGGGTCTTAAAAAATTGATAC |
| tcdA-vef-Fw | GATGGTGCATGGTCAGTTGG |
| tcdA-vef-Rev | GAAGATGGTGATGAGGTGC |
| tcdB-vef-Fw | GACAAGCTGTTAATAAGGC |
| tcdB-vef-Rev | CTGGTAATCCACATAAGCAC |
| YN3-vef-Fw | CATCAAGAAGAGCGACTTCG |
| YN3-vef-Rev | TTCTTTCTATTCAGCACTGTTATGC |
| pyrE-vef-Fw | CAATAATTTTATAACATTAACATGG |
| pyrE-vef-Rev | GTGTTACTTAAAAAATGTAAAT |
| YN4-vef-Fw | CAAGAAGAGCGACTTCGCGGAGCTGG |
| YN4-vef-Rev | CCATTACAGACTTATCCAGGG |
| SpoVT_sgRNA_Fw | TTTTCGTCGACAAGAATAGATGATCTTGGAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGC |
| sgRNA_Rev | GCACCGACTCGGTGCCACTTTTTCAAGTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACGTCGACGAAAA |
| spoVT-AscI-Fw | CCCCGGCGCGCCGAGAAAGATTTAGCAATG |
| spoVT-SOE-Rev | CTTTGAAACAACTACAATTGAACCCTCTTTTGGGATAACTACCCTTCC |
| spoVT-SOE-Fw | GGAAGGGTAGTTATCCCAAAAGAGGGTTCAATTGTAGTTGTTTCAAAG |
| spoVT-AsiSI-Rev | GATGGCGATCGCCTGCAACTTGAGACACAG |
| spoVT-vef-Fw | CGCGGATCCGATGAGTTTTTAAGAGAC |
| spoVT-vef-Rev | CGCGCTCGAGCAAAAGTCTGACCTAGAC |
| PsspA-EcoRI-FW | AGATGAGGAGAATTCGGATAAAAGAGTTCA |
| PsspA-SNAP-SOE-Rev | CATTTCACAATCTTTATCCATGTTGATTACCTTCCTTC |
| SNAP-SOE-Fw | ATGGATAAAGATTGTGAAATGAAGAGAACC |
| SNAPC-XhoI-Rev | CCCCTCGAGTTACCCAAGTCCTGGTTTCCCCAAACG |
aEngineered restriction sites are underlined.