Imperial College London

ProfessorPaulFreemont

Faculty of MedicineDepartment of Medicine

Chair in Protein Crystallography
 
 
 
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Contact

 

+44 (0)20 7594 5327p.freemont

 
 
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Location

 

259Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@inbook{Lai:2018:10.1007/978-1-4939-7795-6_25,
author = {Lai, H-E and Moore, S and Polizzi, K and Freemont, P},
doi = {10.1007/978-1-4939-7795-6_25},
pages = {429--444},
title = {EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.},
url = {http://dx.doi.org/10.1007/978-1-4939-7795-6_25},
year = {2018}
}

RIS format (EndNote, RefMan)

TY  - CHAP
AB - 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.
AU - Lai,H-E
AU - Moore,S
AU - Polizzi,K
AU - Freemont,P
DO - 10.1007/978-1-4939-7795-6_25
EP - 444
PY - 2018///
SP - 429
TI - EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
UR - http://dx.doi.org/10.1007/978-1-4939-7795-6_25
UR - https://www.ncbi.nlm.nih.gov/pubmed/29754244
ER -