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. 2014 Jun 26;5:302. doi: 10.3389/fpls.2014.00302

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Copyright © 2014 Cantos, Francisco, Trijatmiko, Slamet-Loedin and Chadha-Mohanty.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Structure and key features of the pZFN1, pZFN2, and pZFN3 constructs. The vector system is based on the assembly of ZFN expression cassettes (pSAT4.hspP.ZFN and pSAT5.hspP.ZFN), a plant selection expression cassette (pSAT1.hpt), and GUS reporter cassette (pSAT6A.ZFN-TS^*::GUS) onto the plant binary vector pRCS2. Asterisk stands for the modification generated in the cassette. The plasmid carries a plant expression cassette engineered for constitutive expression of a mutated uidA (GUS) gene. A stop (TGA) codon was engineered within the 6-bp spacer of the ZFN target site, leading to premature termination of uidA translation in plant cells. The ZFN QQR recognition site is shown.