Part:BBa_K5159012

Y. lipolytica PEX10 5' homology arm

The sequence is found in the Yarrowia lipolytica genome and used for the knockout of peroxisomal PEX10 gene by homologous recombination (HR). It is located upstream of the genomic PEX10 coding region to be knocked out. This homologous arm together with its downstream counterpart (BBa_K5159013) flanking a selection marker (e.g., LEU2 expression cassette) can locate the PEX10 gene in the genome of Yarrowia lipolytica and help perform a recombination to disable the PEX10 expression, leading to a disrupted peroxisome biogenesis and improved lipid accumulation inside the cells.

As the unconventional yeast Yarrowia lipolytica shows a higher tendency of utilizing non-homologous end-joining (NHEJ) instead of HR for genomic integration, a 1000 bp homologous arm in length is necessary for the efficient knockout to occur.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 196
  Illegal PstI site found at 415
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 196
  Illegal PstI site found at 415
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 251
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 196
  Illegal PstI site found at 415
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 196
  Illegal PstI site found at 415
  
• 1000
  COMPATIBLE WITH RFC[1000]