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. 2010 May 18;38(18):6065–6077. doi: 10.1093/nar/gkq387

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Structure of the NHEJ reporter constructs pEJ and pEJ2. GFP translation is prevented by insertion of an artificial ATG (start codon) into the 5′-untranslated region between CMV promoter and the ORF. The artificial ATG is not in frame with the original ATG (17). Two repeat I-SceI recognition sequences (bold characters) flank the artificial ATG either in direct (pEJ2) or in inverted orientation (pEJ). Simultaneous cleavage of both I-SceI sites leads to pop out of the artificial ATG (middle) leaving either non-compatible ends (pEJ) or fully compatible ends (pEJ2). Repair of the I-SceI-induced DSB by NHEJ restores GFP translation leading to green fluorescence.