Study of ΔdnaA Mutants and their Genomic Position in Synechococcus

1. Fig. S1 a b 0 6 12 24 48 (h) 1021 bp Kmr DnaA CBB 1449 bp dnaA c First selection: transformation Plating on plate with Km Colony PCR-1 Individual ΔdnaA mutants WT M kb 2.69 1.88 1.49 0.93 Cultivation in liquid medium with Km until the cells reach the stationary phase Second selection: Plating on plate with Km Colony PCR-2 ΔdnaA mutants WT 1 2 3 4 5 6 7 8 Segregation: complete (18) incomplete (59) Total 77

2. Fig. S2 a oriC oriC ΔdnaA-1 ΔdnaA-2 b Chr (0826) Plasmid F R P Chr P Chr c R F Chr P Chr P

3. Fig. S3 a b ΔdnaA-1 oriC WT 2328 0302 α-FLAG S. 7942 genome 1943 0680 α-RpoD1 1595 1001 1294 c ΔdnaA-1 ΔdnaA-2 Percent recovery Genomic position

4. Fig. S4 SYTOX Green Auto fluorescence DIC merge WT ΔdnaA-1 ΔdnaA-2

5. Fig. S5 Up: 27 gene Down: 91 gene 10 3 4 2 4 3 2 13 3 48 2 2 2 1 1 3 4 5

6. Fig. S6 ΔdnaATK b a WTTK M Kb 4.26 1021 bp 3.47 Kmr 2.69 1.88 gemome 1.49 1344 bp dnaA c d 792 bp ΔPORTK ΔPORTK ΔPORTK Specr WTTK WTTK WTTK M M M Kb Kb Kb 3 1.88 4.26 1.88 3.47 1.49 1.49 2.69 1 927 bp 1.88 0964 1.49 4 2 1, 2 1, 3 1, 4 Putative oriC region (POR) Containg DnaA box Primer set WTTK e f ΔdnaATK ΔPORTK ΔdnaATK WTTK ΔPORTK dnaN dnaN 0963 α-HA Cell number (X 107) CBB Time (hr) h g ΔdnaATK WTTK pRL２７１ Nmr 1517 1200 1000 900 genome WT; 1280bp dnaA 800 ΔdnaA; 1379bp 700

7. Table S1 Blast search was performed to find the DnaA homologs in plants and algae using Synechococcus DnaA amino acid sequence data as the reference. Indicated data was top 10 in similarity. Upper, plants and algae; lower, cyanobacteria. DnaA protein is not conserved in all plants and photosynthetic prokaryotes, except for cucumber. All the homologous sequences of lower than 23 % indentity in plants and algae are ATPase domains, not DnaA protein-specific domains.

8. Table S2

9. Table S2

10. Table S3