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. 2017 Feb 23;45(11):e94. doi: 10.1093/nar/gkx132

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© The Author(s) 2017. 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 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 2. — TPA optimization based on five-fragments assembly. The columns represent the averages of two independent experiments, and the error bars represent the standard deviations. (A) The zeaxanthin pathway plasmid used for the optimization experiments. CrtE, B, Z, Y, I are the coding regions for the enzymes in the pathway. P represents promoter, R represents RBS and T represents terminator. The double-headed arrows at the bottom denote how the pathway has been broken up. (B and C) Efficiency and fidelity as a function of junction T_M and hybridization T_H. Horizontal axis represents the difference between T_H and T_M. (D) Efficiency as a function of T_H for T_M = 50°C. (E & F) Efficiency and fidelity as a function of annealing protocol. (G & H) Efficiency and fidelity as a function of DNA input amount and backbone:insert ratio. (I & J) Efficiency and fidelity as a function of hybridization time.