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. 2019 Aug 8;47(16):8632–8648. doi: 10.1093/nar/gkz677

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© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 9. — S. thermophilus type II-A spacer integration model. (A) The integrase complex (Cas1–Cas2 bound by a pre-spacer) recognizes 15 bp of the sequences spanning the leader-repeat junction to direct specific integration to the first repeat. (B) Identity of a guanine at position 1 of the repeat facilitates localization of integration to direct the first transesterification attack at the leader-repeat junction, resulting in a half-site intermediate. (C) DNA bending of the repeat sequence initiates the second-site transesterification attack at the repeat–spacer junction measuring 8 nucleotides upstream of a molecular ruler localized near the repeat–spacer junction (Figures 7 and 8). The integration complex additionally relies on a guanine at position 36 to progress to full-site integration of a new spacer into the CRISPR array.