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Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology

Generation of new DNA constructs is an essential process in modern life science and biotechnology. Modular cloning systems based on Golden Gate cloning, using Type IIS restriction endonucleases, allow assembly of complex multipart constructs from reusable basic DNA parts in a rapid, reliable and aut...

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Autores principales: Valenzuela-Ortega, Marcos, French, Christopher
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7889407/
https://www.ncbi.nlm.nih.gov/pubmed/33623824
http://dx.doi.org/10.1093/synbio/ysab003
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author Valenzuela-Ortega, Marcos
French, Christopher
author_facet Valenzuela-Ortega, Marcos
French, Christopher
author_sort Valenzuela-Ortega, Marcos
collection PubMed
description Generation of new DNA constructs is an essential process in modern life science and biotechnology. Modular cloning systems based on Golden Gate cloning, using Type IIS restriction endonucleases, allow assembly of complex multipart constructs from reusable basic DNA parts in a rapid, reliable and automation-friendly way. Many such toolkits are available, with varying degrees of compatibility, most of which are aimed at specific host organisms. Here, we present a vector design which allows simple vector modification by using modular cloning to assemble and add new functions in secondary sites flanking the main insertion site (used for conventional modular cloning). Assembly in all sites is compatible with the PhytoBricks standard, and vectors are compatible with the Standard European Vector Architecture (SEVA) as well as BioBricks. We demonstrate that this facilitates the construction of vectors with tailored functions and simplifies the workflow for generating libraries of constructs with common elements. We have made available a collection of vectors with 10 different microbial replication origins, varying in copy number and host range, and allowing chromosomal integration, as well as a selection of commonly used basic parts. This design expands the range of hosts which can be easily modified by modular cloning and acts as a toolkit which can be used to facilitate the generation of new toolkits with specific functions required for targeting further hosts.
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spelling pubmed-78894072021-02-22 Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology Valenzuela-Ortega, Marcos French, Christopher Synth Biol (Oxf) Research Article Generation of new DNA constructs is an essential process in modern life science and biotechnology. Modular cloning systems based on Golden Gate cloning, using Type IIS restriction endonucleases, allow assembly of complex multipart constructs from reusable basic DNA parts in a rapid, reliable and automation-friendly way. Many such toolkits are available, with varying degrees of compatibility, most of which are aimed at specific host organisms. Here, we present a vector design which allows simple vector modification by using modular cloning to assemble and add new functions in secondary sites flanking the main insertion site (used for conventional modular cloning). Assembly in all sites is compatible with the PhytoBricks standard, and vectors are compatible with the Standard European Vector Architecture (SEVA) as well as BioBricks. We demonstrate that this facilitates the construction of vectors with tailored functions and simplifies the workflow for generating libraries of constructs with common elements. We have made available a collection of vectors with 10 different microbial replication origins, varying in copy number and host range, and allowing chromosomal integration, as well as a selection of commonly used basic parts. This design expands the range of hosts which can be easily modified by modular cloning and acts as a toolkit which can be used to facilitate the generation of new toolkits with specific functions required for targeting further hosts. Oxford University Press 2021-02-02 /pmc/articles/PMC7889407/ /pubmed/33623824 http://dx.doi.org/10.1093/synbio/ysab003 Text en © The Author(s) 2021. Published by Oxford University Press. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Valenzuela-Ortega, Marcos
French, Christopher
Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology
title Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology
title_full Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology
title_fullStr Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology
title_full_unstemmed Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology
title_short Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology
title_sort joint universal modular plasmids (jump): a flexible vector platform for synthetic biology
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7889407/
https://www.ncbi.nlm.nih.gov/pubmed/33623824
http://dx.doi.org/10.1093/synbio/ysab003
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