SYNBRIDGE: A VERSATILE FRAMEWORK FOR BIDIRECTIONAL INTERCONVERSION BETWEEN BIOBRICKS AND GOLDEN GATE ASSEMBLY STANDARDS

Component reusability and interoperability are fundamental to synthetic biology. While BioBricks laid the groundwork for part domestication, Golden Gate-based assembly standards, in particular Modular Cloning (MoClo), revolutionized multipart assembly. Despite widespread adoption, standardized conversion between these two popular frameworks remains challenging due to sequence incompatibilities. While de novo synthesis remains constrained by cost and sequence complexity, conventional PCR-based conversion suffers from off-target binding driven by sequence redundancy in multigene devices, unscalable primer optimization costs, and amplicon size constraints. To address interconversion from an engineering perspective, we present SynBridge: a closed-loop idempotent workflow for bidirectional BioBrick-MoClo conversion built around two core operations, Bri2Mo and Mo2Bri. As a proof of concept, we validated SynBridge by constructing a composite genetic system integrating a BioBrick module for LuxR-mediated RFP expression with a MoClo-assembled device for thermoregulated GFP expression. Quantitative cloning efficiency and functional characterization confirmed a robust and reproducible workflow that operates without requiring ad-hoc solutions. SynBridge provides a versatile engineering tool that ensures backward compatibility, maximizes part reusability, and establishes a standardized interchangeability framework suitable for automated procedures.