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. 2021 Aug 25;4:1007. doi: 10.1038/s42003-021-02541-z

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© The Author(s) 2021

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. 2 — a The schematic representation of the plasmid-based invader assay. Potential PAM sequences (NNN) are introduced upstream of the spacer sequence P23-S1 (orange letters) and then inserted into pWL502 yielding invader plasmids pWL502-NNN. DF50ΔEPS is transformed with invader plasmids and selected on AS-168SY plates without uracil. Only the cells containing the invader plasmid with a non-functional PAM could grow on the plates. No cell growth indicates that the invader plasmid carries a functional PAM which leads to interference and degradation of the invader plasmid. b Relative transformation rates of PAM constructs. Transformation rates are calculated as the number of transformants/μg DNA. The transformation rate of pWL502 is used as the positive control. Data shown for two or three biological replicates. Error bars indicate SDs, n = 3. c Colonies obtained by transformation of 1 μg of plasmid pWL502 or pWL502-TTG.