Part:BBa_K3629019

Yarrowia lipolytica LEU4 gene

2-isopropylmalate (LEU4) coding sequence from Yarrowia lipolytica .

Usage and Biology

2-isopropylmalate synthase is the first enzyme involved in the biosynthesis of leucine. This is a very important enzyme in the pathway as it serves as a regulatory point in the biosynthesis pathway (1). The protein has a leucine binding domain which allows for downregulation of the pathway through negative-feedback inhibition of the enzyme. Expression of this enzyme especially on a strong promoter could be used to up-regulate leucine synthesis and produce a leucine overproducing strain.

Design

Below is the codon-optimized sequence of Yarrowia lipolytica LEU4 for use in any expression construct. However, we have also made our own expression construct BBa_K3629023, designed to allow for optimal expression of LEU4 in Yarrowia lipolytica using the TEfin promoter BBa_K3629001 and the XPR2 terminator BBa_K3629004.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 728
  Illegal BamHI site found at 293
  Illegal XhoI site found at 428
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 799
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 544
  

References

1. [online] https://www.genome.jp/kegg-bin/show_pathway?yli00290 (Accessed October 26, 2020)