Plasmid copy number variation of a modular vector set in Shewanella oneidensis MR-1

To advance synthetic biology approaches that utilize S. oneidensis as host for biotechnology applications, we have investigated the variation in plasmid copy number of a modular vector set resulting from distinct origins of replication under different conditions. The replicons yielded a ≈9X-fold range for plasmid copy number variation in S. oneidensis (while the same origins yielded a ≈3X-fold range in Escherichia coli). This provides a sizeable range to control gene expression levels in S. oneidensis for synthetic biology applications. In addition, plasmid harboring the pBBR1 origin resulted in stable copy numbers in S. oneidensis under different conditions (mid-logarithmic, stationary, multi-plasmid). This may enable the realization of synthetic circuits in S. oneidensis where predictable, quantitative behavior is desired (in either single- or double-plasmid contexts).