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. 2020 Aug 6;10:13297. doi: 10.1038/s41598-020-69990-0

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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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SDS-PAGE of the SpyT/-C and SnpT/-C reactions. Left: Reactions of mtDod Spy-/SnpT constructs with seACP-SpyC and/or mClover3-SnpC. Right: Inverse reactions of mtDod SpyC constructs with SpyT-seACP. In all reactions, bands of higher mass representing the conjugation products are observed. As mentioned above, the acidic loading buffer causes the appearance of double bands (mtDod SpyC constructs) and smearing bands (seACP-SpyC-H8) for some constructs.