Next-generation vaccines and therapies in development, including those based on DNA, RNA, and viral vectors, are all overwhelmingly manufactured using plasmid DNA (pDNA). The traditional method for the large-scale production of pDNA involves fermentation in E. coli strains, which is a laborious, time-consuming, and expensive process, making plasmid production a bottleneck and potential inhibitor of growing success for next-generation therapies. Cell-free routes to synthetic DNA may address the problem.