Part:BBa_K5159020

LEU2 marker expression cassette

The LEU2 marker expression cassette, expressing LEU2 gene with its native promoter and terminator in Yarrowia lipolytica, is usually transformed together with other gene(s) as a selective marker. In a strain (e.g., Yarrowia lipolytica Po1f) knocked out with native LEU2 gene and thus unable to synthesize essential amino acid leucine, presence of LEU2 expression can make the transformed cells selectively grow on leucine- auxotrophic YNB plates. While untransformed cells cannot grow due to auxotrophy.

In order to repeatedly integrate multiple genes into Yarrowia lipolytica, we will release the LEU2 marker expression cassette with the Cre-loxP system, so the LEU2 expression cassette is flanked with 2 loxP sites by its design.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 35
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 35
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 35
  Illegal BglII site found at 2267
  Illegal XhoI site found at 981
  Illegal XhoI site found at 1010
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 35
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 35
  Illegal NgoMIV site found at 123
  Illegal AgeI site found at 1595
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 641
  Illegal BsaI site found at 924
  Illegal BsaI.rc site found at 839
  Illegal SapI site found at 1392