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EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology

Lai, Hung-En and Moore, Simon J. and Polizzi, Karen and Freemont, Paul (2018) EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology. In: Braman, Jeffrey Carl, ed. Synthetic Biology: Methods and Protocols. Springer. ISBN 978-1-4939-7794-9. E-ISBN 978-1-4939-7795-6. (doi:10.1007/978-1-4939-7795-6_25) (KAR id:69389)

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https://doi.org/10.1007/978-1-4939-7795-6_25

Abstract

Development of advanced synthetic biology tools is always in demand since they act as a platform technology to enable rapid prototyping of biological constructs in a high-throughput manner. EcoFlex is a modular cloning (MoClo) kit for Escherichia coli and is based on the Golden Gate principles, whereby Type IIS restriction enzymes (BsaI, BsmBI, BpiI) are used to construct modular genetic elements (biological parts) in a bottom-up approach. Here, we describe a collection of plasmids that stores various biological parts including promoters, RBSs, terminators, ORFs, and destination vectors, each encoding compatible overhangs allowing hierarchical assembly into single transcription units or a full-length polycistronic operon or biosynthetic pathway. A secondary module cloning site is also available for pathway optimization, in order to limit library size if necessary. Here, we show the utility of EcoFlex using the violacein biosynthesis pathway as an example.

Item Type: Book section
DOI/Identification number: 10.1007/978-1-4939-7795-6_25
Uncontrolled keywords: Synthetic biology; Golden gate; MoClo; Violacein; DNA assembly
Subjects: Q Science
Q Science > QR Microbiology
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Simon Moore
Date Deposited: 05 Oct 2018 14:48 UTC
Last Modified: 22 Jan 2020 04:11 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/69389 (The current URI for this page, for reference purposes)
Moore, Simon J.: https://orcid.org/0000-0002-1968-206X
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