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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 4. — (a) Chromosomal insertion positions. Fragments were inserted into six positions distributed symmetrically about the E. coli chromosomal origin of replication oriC. Insertion positions are between the marked genes at each position (dots). (b) Verification of chromosomal insertions. Colony PCR across insertion junctions for insertion of the P_lacI-lacI:venus:T1-neo cassette (IvT-neo; 2.1-kb band) into the nth-ydgR position and the P_LlacO1-lacZ-cat cassette (O1Z-cat; 5-kb band) at each site. The insertion of both the IvT-neo and O1Z-cat cassettes into the nth-ydgR position was accomplished by a single 7-kb insertion bearing the neo marker. Lanes 2–8 are MG1655 Δlac negative controls in the following order: lane 2: nth IvT-neo; lanes 3–8: O1Z in alphabetical order of insertion positions, corresponding to positive insertion lanes 10–20.