CyanOperon: An Operon Building Expansion for the CyanoGate MoClo Toolkit
Year: 2026
Type: article
Abstract: ABSTRACT Operons are gene clusters controlled by a single promoter that enable coordinated translation from a single messenger RNA. Here we describe an expansion of the CyanoGate MoClo toolkit to assemble synthetic operons. The versatile CyanOperon system includes two Level 0 acceptor vectors for building interchangeable promoter-ribosome bind site (RBS) combinations and 15 Level 1 acceptor vectors for the hierarchical assembly and expression of up to six genes within a single operon. The system also allows for operon assembly into a self-replicating vector or for chromosomal integration by homologous recombination. To showcase CyanOperon, we assembled the violacein biosynthesis pathway as an operon and demonstrated violacein production in Escherichia coli . We then constructed a 20-part RBS library to examine how spacer length between the Shine–Dalgarno sequence and start codon affects translation in E. coli and the model cyanobacterium Synechocystis sp. PCC 6803. Lastly, we compared the expression of up to three operonic fluorescent markers following chromosomal integration or from a self-replicating vector in E. coli and Synechocystis sp. PCC 6803. The CyanOperon system is publicly available and can be readily integrated with other MoClo systems to accelerate the development of standardized operon assemblies. GRAPHICAL ABSTRACT
Source: ACS Synthetic Biology
Institution: University of Edinburgh
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: A powerful directed-evolution tool for exploitation of chloroplast engineering biology, Engineering cyanobacteria into bio-solar cell factories for scalable carbon capture utilisation and storage, Rewriting The Genetic Code: The Algal Plastome As A Testbed For Basic And Applied Studies, 22BBSRC-NSF/BIO: A synthetic pyrenoid to guide the engineering of enhanced crops
OpenAlex ID: W7155567940