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The Standard European Vector Architecture (SEVA) format released

Víctor de Lorenzo. Centro Nacional de Biotecnología, CSIC. Madrid 28049, Spain. vdlorenzo@cnb.csic.es

The failure of the pioneers of recombinant DNA technologies to implement standards for the construction and nomenclature of plasmids and other genetic tools has resulted in decades of chaotic development of cloning vectors and other molecular assets. But the onset of Synthetic Biology has made apparent the necessity of fixed formats for vector organization and designation. In addition, the vast majority of genetic engineering efforts are made in E. coli. This is an excellent host for the physical assembly of DNA parts into devices and modules but often, this organism is not the optimal platform for the deployment of the designed properties in a biotechnological setting, e.g., a bioreactor or released to the environment. Many attempts to expand the vector toolbox beyond enteric bacteria have been made in the past decades but their molecular architectures have been generally capricious and largely dependent on the immediate application for which they were created. While most of these issues may be ultimately resolved through in vitro assembly of DNA segments and even synthesis of entire genomes á la carte, the immediate future still needs better molecular tools for deployment of the engineered features of interest in the most advantageous carrier. The BioBrick and BglBrick standards, popularized through the iGEM competition, are useful platforms for the recurrent physical assembly of DNA segments, but they do not tackle the issue of functionality or deployment of the constructs. In this context the initiative Standard European Vector Architecture (SEVA) was launched time ago for helping participants of Syn Bio Projects of the 6th and 7th Framework Programmes to access a coherent resource of molecular tools that are subject to a concise, minimalist and standardized format and nomenclature. Importantly, these tools are compatible with old and new cloning and DNA assembly methods. The resulting set of plasmid and transposon vectors, which are available in the http://seva.cnb.csic.es database, is composed of a number of synthetic, interchangeable and reusable functional modules that include a broad-host-range origin of replication, antibiotic markers, expression systems and reporter genes that are punctuated by terminators and gadget-insertion sites (Fig. 1). The first collection of these formatted vectors was intended for analysis and assembly of engineered phenotypes in a suite of Gram-negative bacteria, but they are currently being enriched and expanded towards Gram-positives with contributions from the ETH Zürich, the TU Berlin, U Groningen, the Imperial College London and many others. We advocate that the SEVA format may become a fundamental reference to speed up the ease of biosystems engineering beyond laboratory applications. 

Figure 1. Organization of SEVA plasmids. Vectors that follow the format are formed by three variable modules: a cargo, a replication origin and an antibiotic marker. Rare restriction sites used to change the functional modules are shown. Segments are separated by three permanent regions, which are shared by all vectors, the T0 and T1 transcriptional terminators and the oriT conjugation origin.