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. 2014 Nov 5;42(21):13440–13451. doi: 10.1093/nar/gku1082

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© The Author(s) 2014. 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. — Overview of mammalian circuit assembly (mMoClo). (A) mMoClo is based on a library of empty backbones used to assemble parts (Parts Positioning Vectors), transcription units (Transcription Unit Positioning Vectors) and complex multi-unit circuits (Destination Vector) to be integrated into the landing pad. The Parts Library is made of sequence-verified basic genetic parts (Insulators, Promoters, 5′ UTR, Genes, 3′UTR and polyA sequences). (B) One-step creation of an integration vector carrying BxB1 attB site and promoterless selection marker. (C) One-step assembly of different functional transcription units in specific position vectors. (D) One-step assembly of a multi-unit genetic circuit by combining the desired transcription units, the corresponding linker vector and the previously built integration vector carrying the promoter-trap cassette. Details of the overhangs used in the assembly process are provided in Supplementary Figure S4.