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. 2017 Jul 21;7:6119. doi: 10.1038/s41598-017-06428-0

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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De novo synthesis of lycopene biosynthetic pathway. Fragments were assembled by multiplex assembly using error-depleted oligo pools of various throughputs and half-specific primer pairs. Thirty six (a), 3–4 (b), 6–7 (c), and 17–19 (d) subpools were used in one assembly reaction and 24, 26, 24, and 22 fragments were assembled, respectively. (e) Fragment detection in the assembly (36 subpool/reaction) with full-specific primer pairs. (f) Fragment detection in the assembly (12 subpool/reaction) with half-specific primer pairs. (g) Fragment detection in the assembly (12 subpool/reaction) with full-specific primer pairs. (h) Operons assembled by Gibson assembly. M1, 20 bp DNA ladder (TaKaRa Bio); M2, 2 K plus II DNA ladder (TaKaRa Bio). Red arrows indicate target fragments. OLS, oligo subpool; F, fragment.