Journal
NUCLEIC ACIDS RESEARCH
Volume 40, Issue 12, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gks236
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Funding
- Bauer Fellows Program
- ETH Zurich
- National Institutes of Health [1R01CA155320-01]
- National Institute of General Medical Sciences Grant for Centers of Systems Biology
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Synthetic biology applications call for efficient methods to generate large gene cassettes that encode complex gene circuits in order to avoid simultaneous delivery of multiple plasmids encoding individual genes. Multiple methods have been proposed to achieve this goal. Here, we describe a novel protocol that allows one-step cloning of up to four gene-size DNA fragments, followed by a second assembly of these concatenated sequences into large circular DNA. The protocols described here comprise a simple, cheap and fast solution for routine construction of cassettes with up to 10 gene-size components.
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