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. 2018 Aug 1;9:3004. doi: 10.1038/s41467-018-05109-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Characterization of the Kai copy-number tunable strain. a A theophylline riboswitch regulates translational efficiency of all three kai genes, and transcriptional regulation of kaiA is controlled by an IPTG-inducible promoter. Clock state is reported by EYFP-SsrA expressed from the kaiBC promoter. b Theophylline regulates translation by freeing the ribosome binding site upstream of each kai gene. c Kai copy numbers plotted as a function of theophylline concentration with 1 μM IPTG (solid line), and Kai copy numbers in wild-type cells (dotted line). Vertical error bars or shaded area indicate standard error of the mean from three replicates. d Colony-level oscillations detected with a bioluminescent reporter in the copy number tunable strain with 1 μM IPTG and various theophylline concentrations