Bailey Heron, Steve Petrovski, Galain C. Williams, Dena Lyras
Over time, numerous legacy bacterial isolates and mobile genetic elements (MGEs) have been systematically lost as physical strain collections have been passed down across academic generations. This ongoing attrition has significantly restricted the ability of contemporary scientists to reproduce historical findings or build upon foundational research. However, recent advancements in high fidelity commercial DNA synthesis, coupled with modern recombinant DNA technologies, now enable researchers to precisely reconstruct lost MGEs based entirely on digital sequence records. Here, we demonstrate the de novo physical reconstitution of PRR1, a plasmid-dependent, single-stranded RNA bacteriophage that has evidently become lost across many global culture repositories. Comprehensive genetic, phenotypic, and ultrastructural characterisation confirmed that the synthetic PRR1 virions exhibit biological and structural traits completely identical to the historically documented virology of the wild-type phage. Ultimately, this study highlights how the synthesis and operational rebooting of lost MGEs is now readily achievable within exceptionally short time frames, transforming public sequence databases into active toolkits for evolutionary and antimicrobial research.