Reagents and tools table
| Reagent/resource | Reference or source | Identifier or catalog number |
|---|---|---|
| Experimental models | ||
| Synechococcus sp. PCC7942 kaiABCΔ | Obtained from Johnson CH Lab | N/A |
| Synechococcus sp. PCC7942 strain harbouring a luxAB reporter (PkaiBC::luxAB) | Obtained from Johnson CH Lab | N/A |
| Synechococcus sp. PCC7942 kaiABCΔ: kaiABC | This study | N/A |
| Synechococcus sp. PCC7942 kaiABCΔ: ankaiABC | This study | N/A |
| PCR primers | ||
| Forward primer for construction of cyanobacterial transformation plasmids pKai-TR-ankaiABC and pKai-TR-ankaiBC (V-A-homo-F) | This study | 5’-cagcggcggaggaactctttga-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmids pKai-TR-ankaiABC and pKai-TR-ankaiBC (V-A-homo-R) | This study | 5’-tcaaagagttcctccgccgctg-3’ |
| Forward primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (V-A-F) | This study | 5’-aaaggtaaaggaggtcttaagctcgg-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmids pKai-TR-ankaiABC and pKai-TR-ankaiBC (V-A-R) | This study | 5’-gcagtcgctcctgtcaggac-3’ |
| Forward primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (V-B-F) | This study | 5’-ctgtcgttaactgctttgttggtact-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (V-B-R) | This study | 5’-acgcagatcaacggggtagca-3’ |
| Forward primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (V-C10-F) | This study | 5’-agctttatgcttgtaaaccgttttgt-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (V-C10-R) | This study | 5’-cttaaaagagggtgaagtcaggtagt-3’ |
| Forward primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (An-A10-F) | This study | 5’-gtcctgacaggagcgactgcatgttatctgccttaacgatct-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (An-A10-R) | This study | 5‘-ttaagacctcctttacctttttaagaagaaacctcagagtc-3’ |
| Forward primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (An-B10-F) | This study | 5’-gctaccccgttgatctgcgtatgagcccgctgcgcaaaaccta-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (An-B10-R) | This study | 5’- aacaaagcagttaacgacagttacggatcgctatccggc-3’ |
| Forward primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (An-C10-F) | This study | 5’-tgacttcaccctcttttaagatgaccagcctggcggaacat-3’ |
| Reverse primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiABC (An-C10-R) | This study | 5’-cggtttacaagcataaagctttagctttccggttctttttcc-3’ |
| Forword primer for construction of cyanobacterial transformation plasmid pKai-TR-ankaiBC (anBC-F) | This study | 5’-gacaggagcgactgcaaaggtaaaggaggtcttaagctcgg-3’ |
| Reverse primer of KaiA for genotyping (cpWt1-F) | This study | 5’-tgaaccagccaaagaaca-3’ |
| Reverse primer of KaiA for genotyping (cpWt1-R) | This study | 5’-ggaacatcggcaaagaaa-3’ |
| anKaiB+anKaiC for genotyping (anAn1-F) | This study | 5’-gaactgctgctggatgat-3’ |
| anKaiB+anKaiC for genotyping (anAn1-R) | This study | 5’-gatccaccagtttctggca-3’ |
| Recombinant DNA | ||
| pGEX-6P-1 | Addgene | Cat#: 27-4597-01 |
| pGEX-6P-1-kaiC | This study | N/A |
| pGEX-6P-1-kaiA | This study | N/A |
| pGEX-6P-1-kaiB | This study | N/A |
| pGEX-6P-1-anKaiC | This study | N/A |
| pLou3-ankaiA | This study | N/A |
| pGEX-6P-1-ankaiB | This study | N/A |
| pGEX-6P-1-kaiC-V118A | This study | N/A |
| pGEX-6P-1-kaiC-S229T | This study | N/A |
| pGEX-6P-1-kaiC-T240N | This study | N/A |
| pGEX-6P-1-KaiC-S229T-T240N | This study | N/A |
| pGEX-6P-1-psp | This study | N/A |
| pRK793 | Addgene | Cat#: 8827 |
| pKai-TR-kaiABC | Obtained from Johnson CH Lab | Nishimura et al, 2002 |
| pKai-TR-ankaiABC | This study | N/A |
| pKai-TR-ankaiBC | This study | N/A |
| Antibodies | ||
| Rabbit anti-KaiC 1:5000 for immunoblotting | This study | N/A |
| Goat Anti-Rabbit IgG (H + L)-HRP Conjugate | Bio-Rad | Cat#:170-6515 |
| Chemicals, enzymes and other reagents | ||
| TransStart® Taq DNA Polymerase | TransGen Biotech | AP141-13 |
| FastPure EndoFree Plasmid Midi Kit | Vazyme | DC205-01 |
| BG11 | Hopebio | HB8793 |
| Agar Powder | Hopebio | HB8274 |
| Chloramphenicol | Sangon Biotech | A100230 |
| Spectinomycin | Sangon Biotech | A600901 |
| Kanamycin | Sangon Biotech | A600286 |
| Agar | Sangon Biotech | A505255 |
| Tryptone | Oxoid | LP0042B |
| Yeast extract | Oxoid | LP0021 |
| NaCl | Sangon Biotech | A501218 |
| Tris | Sangon Biotech | A600194 |
| IPTG Dioxane Free | Sangon Biotech | A600168 |
| Bam HI | New England Biolabs | R3136V |
| Sac I | New England Biolabs | R3156S |
| Glutathione reduced | Sangon Biotech | A600229 |
| Glutathione Sepharose 4B | Coolaber | CS20421 |
| ATP | Sigma-Aldrich | A1852 |
| ADP | Sigma-Aldrich | A2754 |
| Software | ||
| SnapGene | SnapGene Software | https://www.snapgene.com/ |
| ImageJ | National Institutes of Health | https://imagej.nih.gov/ij/ |
| R software | R Foundation Statutes | https://www.r-project.org/ |
| GraphPad Prism 8 | Graphpad Software | https://graphpad.com |
| CRYSON | Svergun et al, 1998 | https://www.embl-hamburg.de/biosaxs/cryson.html |
| SasView | SasView Software | https://www.sasview.org/ |
| Other | ||
| anKaiC structure | This study | PDB:8JON |
| KaiC structure | Swan et al, 2022 | PDB:7S67 |