PT  - JOURNAL ARTICLE
AU  - Ravendran Vasudevan
AU  - Grant A.R. Gale
AU  - Alejandra A. Schiavon
AU  - Anton Puzorjov
AU  - John Malm
AU  - Michael D. Gillespie
AU  - Konstantinos Vavitsas
AU  - Valentin Zulkower
AU  - Baojun Wang
AU  - Christopher J. Howe
AU  - David Lea-Smith
AU  - Alistair J. McCormick
TI  - CyanoGate: A Golden Gate modular cloning suite for engineering cyanobacteria based on the plant MoClo syntax
AID  - 10.1101/426700
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 426700
4099  - http://biorxiv.org/content/early/2018/10/02/426700.short
4100  - http://biorxiv.org/content/early/2018/10/02/426700.full
AB  - Recent advances in synthetic biology research have been underpinned by an exponential increase in available genomic information and a proliferation of advanced DNA assembly tools. The adoption of plasmid vector assembly standards and parts libraries has greatly enhanced the reproducibility of research and exchange of parts between different labs and biological systems. However, a standardised Modular Cloning (MoClo) system is not yet available for cyanobacteria, which lag behind other prokaryotes in synthetic biology despite their huge potential in biotechnological applications. By building on the assembly library and syntax of the Plant Golden Gate MoClo kit, we have developed a versatile system called CyanoGate that unites cyanobacteria with plant and algal systems. We have generated a suite of parts and acceptor vectors for making i) marked/unmarked knock-outs or integrations using an integrative acceptor vector, and ii) transient multigene expression and repression systems using known and novel replicative vectors. We have tested and compared the CyanoGate system in the established model cyanobacterium Synechocystis sp. PCC 6803 and the more recently described fast-growing strain Synechococcus elongatus UTEX 2973. The system is publicly available and can be readily expanded to accommodate other standardised MoClo parts.