Part:BBa_K1484317:Design
4-Hydroxybenzalacetone Generator
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 2350
Illegal BamHI site found at 2696
Illegal XhoI site found at 3978 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 3282
Illegal AgeI site found at 979 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2409
Illegal SapI site found at 961
Design Notes
It consists of the enzymes 4-coumarate:coA ligase (from N. tabacum; BBa_K1484115) and benzalacetone synthase (from Rheum palmatum, Chinese rhubarb; BBa_K1484103), under the control of an arabinose inducible promoter.
The two genes are transcribed into a polycistronic operon.
The first RBS is BBa_J34801, the second is a synthetic RBS calculated by the Salis Lab RBS calculator which can be found at https://salis.psu.edu/software/
Source
The cds of the 4-coumarate:coA ligase was kindly sent by Jules Beekwilder and Esmer Jongedijk of Wageningen UR in the Netherlands, and that of benzalacetone synthase by Ikuro Abe and Takahiro Mori of The Graduate School of Pharmaceutical Sciences at the University of Tokyo.
References
Jules Beekwilder et al, Microbial production of natural raspberry ketone, Biotechnol. J. 2007, 2, 1270–1279
Yoshihiko Shimokawa, Hiroyuki Morita, and Ikuro Abe, /Benzalacetone synthase, Front Plant Sci. 2012; 3:57.
Yoshihiko Shimokawa, Hiroyuki Morita, and Ikuro Abe Structure-based engineering of benzalacetone synthase, Bioorganic & medicinal chemistry letters 09/2010; 20(17):5099-103.