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. 2021 Jan 11;49(3):1769–1783. doi: 10.1093/nar/gkaa1167

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© The Author(s) 2021. 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-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — Modular genome engineering strategy. Schematic representation of the design and construction of neochromosomes (A), as well as screening based on fluorescence, size and sequencing (B). The data in panel B represent the results pertaining to the assembly of the 100 kb neochromosome in strain IMF6. The red bar in the top left of the fluorescence microscopy pictures (40x) represents 10 μm. The CHEF gel used to determine the size of the neochromosomes, contains two transformants (#3 and #4) of the 100 kb neochromosome transformation. The neochromosomes are either undigested (undig.) by I-SceI and therefore in their circular form, or in-plug linearized by I-SceI digestion (dig.) and therefore in linear form, which can be visualised on CHEF gel.