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Data S1. Materials and methods.

Fig. S1. Effect of various liquid media on S. sahachiroi protoplast formation and regeneration.

Fig. S2. Effect of culture time on S. sahachiroi protoplast formation and regeneration.

Fig. S3. Effects of lysozyme concentration and reaction time on S. sahachiroi protoplast formation and regeneration.

Fig. S4. Effect of recipient/donor ratio on conjugation efficiency.

Fig. S5. Effect of media on sporulation of S. sahachiroi.

Fig. S6. Conjugation and transformation plates.

Fig. S7. PCR confirmation of the in-frame deletion mutant ΔaziU3 and the complementation mutant ΔaziU3::aziU3.

Fig. S8. PFGE analysis of the native linear plasmids in S. sahachiroi ATCC 33158.

Table S1. Bacterial strains and plasmids used in this study.

Table S2. PCR primers used in this study.

Table S3. Sensitivity of S. sahachiroi ATCC 33158 to antibiotics.

Table S4. Effect of culture temperature on protoplast formation and regeneration.

Table S5. Effect of regeneration media on protoplast regeneration.

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