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. Author manuscript; available in PMC: 2016 Apr 26.
Published in final edited form as: Methods Mol Biol. 2015;1311:349–362. doi: 10.1007/978-1-4939-2687-9_23

Fig. 3.

Fig. 3

Primer design and overhang sequences for Golden Gate cloning. (a) BsaI is a type IIS restriction enzyme that cuts outside of the recognition site. (b) The primers (both forward and reverse) contain a BsaI site, a unique 4-nt overhang, and homologous sequence binding to the PCR vector. (c) An orthogonal set of overhangs for Golden Gate cloning. These orthogonal overhangs (A–I) prevent erroneous ligation during the ligation step