Protein_Domain

Part:BBa_K4895000:Design

Designed by: Patrick Jiang, Rori Hoover   Group: iGEM23_ASU   (2023-09-14)


Salmonella Enterica LT2 strain fadA (3-ketoacyl-CoA thiolase)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Unknown
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Many of our considerations dealt with the length of and the possible toxicity of the enzyme to bacteria. It is found that fadA comes from a family of Beta-oxidative genes under the fadR regulon. The fadR regulon works as an apoprotein double repressor for both the fad family and fab family.

The fab family consists of other metabolically active enzymes, that instead oxidate unsaturated long chain fatty acids.

In the presence of acyl-coA's, the fadR apoprotein will unbind from fad family genes and bind to the UTR of fab family genes. However, without acyl-coA, fadR always binds to the UTR of fad family, and does not bind to fab family. As such, it is important to consider that the heavy regulation of native fad genes may become a hinderance when beta oxidation needs to be constitutively expressed. This sequence only contains the coding region, and avoids the regulation by fadR.


Source

Genomic sequence of Salmonella Enterica LT2 strain. Member of the fad family of beta-oxidative enzymes. This sequence is adapted from amino acid sequences before being codon-optimized to fit the transcriptional needs of E.Coli. The codon optimization was performed through Benchling.

References