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. 2010 Jan 4;38(6):e92. doi: 10.1093/nar/gkp1193

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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 5. — (a) Verification of large chromosomal insertions. Colony PCR of 16 randomly picked colonies obtained from insertion of IvT-O1Z-neo between atpI and gidB. Lane 2: MG1655 Δlac pTKRED negative control atpI proximal junction. Lane 3: MG1655 Δlac pTKRED negative control gidB proximal junction. Lanes 5–20 and 22–37 are IvT-O1Z-neo cassette integrants. Lanes are alternating pairs of atpI proximal junctions (2.1-kb bands; lacI:venus:T1 amplified) and gidB proximal junctions (5-kb bands; lacZ-neo amplified) for each clone. (b, c) Alignment of insertion junction sequences obtained from first eight clones shown in (a). (b) Junction proximal to atpI. (c) Junction proximal to gidB. Sequences of the flanking chromosomal regions, 25-bp landing pad regions, and the inserted fragment are labeled and indicated by a black, red or purple underscore, respectively. Mismatches to the expected sequence are highlighted in blue.