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The pGinger family of expression plasmids

Allison N. Pearson, Mitchell G. Thompson, Liam D. Kirkpatrick, Cindy Ho, Khanh M. Vuu, Lucas M. Waldburger, Jay D. Keasling, Patrick M. Shih
doi: https://doi.org/10.1101/2023.01.23.524619
Allison N. Pearson
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
2Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
3Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA
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Mitchell G. Thompson
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
4Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA
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Liam D. Kirkpatrick
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
4Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA
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Cindy Ho
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
2Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
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Khanh M. Vuu
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
4Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA
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Lucas M. Waldburger
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
2Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
5Department of Bioengineering, University of California, Berkeley, California, USA
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Jay D. Keasling
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
2Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
6Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, USA
7The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Denmark
8Center for Synthetic Biochemistry, Institute for Synthetic Biology, Shenzhen Institutes for Advanced Technologies, Shenzhen, China
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  • For correspondence: jdkeasling@lbl.gov pmshih@lbl.gov
Patrick M. Shih
1Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA
3Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA
4Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA
9Innovative Genomics Institute, University of California, Berkeley, CA
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  • For correspondence: jdkeasling@lbl.gov pmshih@lbl.gov
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Abstract

The pGinger suite of expression plasmids comprises 43 plasmids that will enable precise constitutive and inducible gene expression in a wide range of gram-negative bacterial species. Constitutive vectors are composed of 16 synthetic constitutive promoters upstream of RFP, with a broad host range BBR1 origin and a kanamycin resistance marker. The family also has seven inducible promoters (Jungle Express, NahR, XylS, RhaS, Lac, LacUV5, and TetR) controlling RFP expression on BBR1/kanamycin plasmid backbones. For four of these inducible promoters (Jungle Express, NahR, Lac, and TetR), we created variants that utilize the RK2 origin and spectinomycin or gentamicin selection. Relevant RFP expression and growth data have been collected in the model bacterium Escherichia coli as well as Pseudomonas putida. All pGinger vectors are available via the Joint BioEnergy Institute (JBEI) Public Registry.

Competing Interest Statement

J.D.K. has financial interests in Amyris, Ansa Biotechnologies, Apertor Pharma, Berkeley Yeast, Demetrix, Lygos, Napigen, ResVita Bio, and Zero Acre Farms.

Footnotes

  • https://public-registry.jbei.org/folders/771

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 23, 2023.
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The pGinger family of expression plasmids
Allison N. Pearson, Mitchell G. Thompson, Liam D. Kirkpatrick, Cindy Ho, Khanh M. Vuu, Lucas M. Waldburger, Jay D. Keasling, Patrick M. Shih
bioRxiv 2023.01.23.524619; doi: https://doi.org/10.1101/2023.01.23.524619
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The pGinger family of expression plasmids
Allison N. Pearson, Mitchell G. Thompson, Liam D. Kirkpatrick, Cindy Ho, Khanh M. Vuu, Lucas M. Waldburger, Jay D. Keasling, Patrick M. Shih
bioRxiv 2023.01.23.524619; doi: https://doi.org/10.1101/2023.01.23.524619

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