Part:BBa_M36344
Actuator for Grape Scent
This part is an actuator for the aroma normally associated with grape soda and consists of a RBS, CDS, and transcriptional terminator. The RBS uses a bicistronic design architecture, which facilitates linkage to promoter regions and allows for similar levels of protein expression across multiple environments. Our coding DNA sequence produces the AMAT enzyme (Anthraniloyl-coenzymeA: methanol acyltransferase), which catalyzes the reaction between anthraniloyl coA and methanol to produce methyl anthranilate, which is an aromatic compound responsible for the grape smell. The transcriptional terminator was specifically chosen as it is complementary to the RBS used.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1274
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1274
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1274
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1274
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1274
Illegal NgoMIV site found at 1048
Illegal AgeI site found at 578 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The amino acid sequence for the AMAT gene was found in the UniProt database, originally from Wang, J., De Luca, V. 2005. We then took the amino acid sequence and ran it through OPTIMIZER, referenced below, to arrive at our nucleotide sequence. A base pair had to be modified from this codon translation to avoid palindromic sequences. Specifically, base pair 1078 of our sequence was changed from a C to a G.
Wang, J., De Luca, V. “The biosynthesis and regulation of biosynthesis of Concord grape fruit esters, including ‘foxy’ methylanthranilate.” The Plant Journal. 2005. Vol. 44, pp. 606-619.
Puigbò, P., Guzmán, E., Romeu A., and Garcia-Vallvé S. “OPTIMIZER: a web server for optimizing the codon usage of DNA sequences.” Nucleic Acids Res. July, 2007. Issue 35. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1933141/
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