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. 2014 Jul 23;5:369. doi: 10.3389/fmicb.2014.00369

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Copyright © 2014 Karas, Wise, Sun, Venter, Glass, Hutchison, Smith and Suzuki.

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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Restoration of the replication rate of JCVI-syn1.0 ΔL by inserting complementation fragment 1 (containing nusG) via the marker-less approach. The line for JCVI-syn1.0 (blue) and the line for rescued clone ML1 (green) overlap. Doubling times calculated from PicoGreen fluorescence (RFU) are 62 min for JCVI-syn1.0 (y = 54.732e^0.0112x, R² = 0.9951), 64 min for complemented clone ML1 (y = 58.374e^0.0108x, R² = 0.9951), and 116 min for JCVI-syn1.0 ΔL (y = 31.425e^0.006x, R² = 0.983).