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Recombinant DNA technology has revolutionized biomedical research with continual innovations advancing the speed and throughput of molecular biology. Nearly all these tools, however, are reliant on Escherichia coli as a host organism, and its lengthy growth rate increasingly dominates experimental time. Here we report the development of Vibrio natriegens, a free-living bacteria with the fastest generation time known, into a genetically tractable host organism. We systematically characterize its growth properties to establish basic laboratory culturing conditions. We provide the first complete Vibrio natriegens genome, consisting of two chromosomes of 3,248,023 bp and 1,927,310 bp that together encode 4,578 open reading frames. We reveal genetic tools and techniques for working with Vibrio natriegens. These foundational resources will usher in an era of advanced genomics to accelerate biological, biotechnological, and medical discoveries.

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