Difference between revisions of "Part:BBa K3629019"

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===Usage and Biology===
 
===Usage and Biology===
  
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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. The protein has a leucine binding domain which allows for down regulation 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.
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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.
  
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See [https://parts.igem.org/Part:BBa_K3629023 BBa_K3629023] for a functional high expression construct using this coding sequence.
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===Design===
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Below is the codon-optimized sequence of <i>Yarrowia lipolytica</i> LEU4 for use in any expression construct. However, we have also made our own expression construct [https://parts.igem.org/Part:BBa_K3629023 BBa_K3629023], designed to allow for optimal expression of LEU4 in <i>Yarrowia lipolytica</i> using the TEfin promoter [https://parts.igem.org/Part:BBa_K3629001 BBa_K3629001] and the XPR2 terminator [https://parts.igem.org/Part:BBa_K3629004 BBa_K3629004].
  
 
===Sequence and Features===
 
===Sequence and Features===
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===References===
 
===References===
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1. KEGG PATHWAY: Valine, leucine and isoleucine biosynthesis. (n.d.). Retrieved October 26, 2020, from https://www.genome.jp/kegg-bin/show_pathway?yli00290
  
 
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Latest revision as of 01:50, 28 October 2020


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. KEGG PATHWAY: Valine, leucine and isoleucine biosynthesis. (n.d.). Retrieved October 26, 2020, from https://www.genome.jp/kegg-bin/show_pathway?yli00290