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. 2020 Mar 23;11:1529. doi: 10.1038/s41467-020-15272-2

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© The Author(s) 2020

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. 4 — a Genetic circuits for the arabinose- and rhamnose-induced expression of split mCherry-intein N- and C-terminal chimeric proteins, respectively, from two different plasmids (top, left), and schematics of protein trans-splicing, exemplified using M86 split at site S2 (bottom). The AND gate behavior (top, right) is achieved only when both inducers are present. Promoters are represented by straight angle arrows and RBS by black semi-circles. Split mCherry halves are shown in gray, M86 intein N-terminal in blue, M86 intein C-terminal in red, the added junction sequence residues (VDA and SDL) in green and the spliced mCherry in purple. b–h Background subtracted fluorescence (Fluo.) of E. coli cells harboring the genetic circuits for split mCherry-intein chimeric proteins, in the absence (−) or presence (+) of inducers. i Genetic circuits for the arabinose- and rhamnose-induced expression of split ECF^mut-split intein N- and C-terminal chimeric proteins, respectively, from two different plasmids (top, left), and schematics of protein trans-splicing, exemplified using ECF16^mut and the SspGyrB intein (bottom). Split ECF16^mut halves are shown in dark pink and SspGyrB halves in light pink. After splicing, the reconstituted ECF16^mut can activate its cognate promoter (P₁₆) and the fluorescent reporter is expressed. j–l Background subtracted fluorescence (Fluo.) of E. coli cells harboring the genetic circuits for the expression of the chimeric split ECF16^mut × SspGyrB halves and expression of the mCherry reporter from P₁₆ (j), the chimeric split ECF17^mut × NrdJ-1 and expression of the mTagBFP reporter from P₁₇ (k) or the chimeric split ECF20^mut × M86 and expression of the GFP reporter from P₂₀ (l), in the absence (−) or presence (+) of inducers. In all the cases, fluorescence was measured 6 h after induction and it was normalized to the cell density (OD₆₀₀). Bars in b–h and j–l represent the mean of three biological replicates (n = 3) and error bars correspond to s.d. Source data are provided as a Source Data file.