SEVA Linkers: A Versatile and Automatable DNA Backbone Exchange Standard for Synthetic Biology

Research output: Contribution to journalJournal article – Annual report year: 2016Researchpeer-review

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DNA vectors serve to maintain and select recombinant DNA in cell factories, and as design complexity increases, there is a greater need for well-characterized parts and methods for their assembly. Standards in synthetic biology are top priority, but standardizing molecular cloning contrasts flexibility, and different researchers prefer and master different molecular technologies. Here, we describe a new, highly versatile and automatable standard “SEVA linkers” for vector exchange. SEVA linkers enable backbone swapping with 20 combinations of classical enzymatic restriction/ligation, Gibson isothermal assembly, uracil excision cloning, and a nicking enzyme-based methodology we term SEVA cloning. SEVA cloning is a simplistic one-tube protocol for backbone swapping directly from plasmid stock solutions. We demonstrate the different performance of 30 plasmid backbones for small molecule and protein production and obtain more than 10-fold improvement from a four-gene biosynthetic pathway and 430-fold improvement with a difficult-to-express membrane protein. The standardized linkers and protocols add to the Standard European Vectors Architecture (SEVA) resource and are freely available to the synthetic biology community.
Original languageEnglish
JournalA C S Synthetic Biology
Volume5
Issue number10
Pages (from-to)1177–1181
Number of pages5
ISSN2161-5063
DOIs
Publication statusPublished - 2016
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • Synthetic biology standards, Plasmid backbone exchange, Standard parts characterization, Cell factory design

ID: 122264955