Composite

Part:BBa_M36344

Designed by: Chris Aholt, Arjun Bhargava, Nikhil Bhargava   Group: Stanford BIOE44 - S11   (2012-12-04)

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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1274
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1274
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1274
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1274
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1274
    Illegal NgoMIV site found at 1048
    Illegal AgeI site found at 578
  • 1000
    COMPATIBLE WITH RFC[1000]

Circular specification. Registry curation 5-4-2014

This part was defined as a composite part including itself . This definition is recursive and not valid. The designer should define a basic part with only the parts sequence and then define a separate composite part with the enclosing parts. The current part is just many copies of the mCherry part

We have redefined it as a basic part.


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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