Part:BBa_K4879003

Yarrowia lipolytica HygR marker

Resistance marker gene for Hygromycin B, designed for expression for Yarrowia lipolytica.

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Usage and Biology

Due to the limited number of auxotrophic markers available for Yarrowia lipolytica, we had to use antibiotic-resistance markers for the selection of possible attempted deletion in our chassis. In order to use the selection marker, we had to use 200µg/ml of hygromycin B in our selection plates.

Hygromycin B tends to kill cells by interfering in polypeptide synthesis, making the organism unable to synthesise proteins. The selection marker gene, which codes for a phosphotransferase, confers resistance by modifying the hygromycin B drug molecule; making it ineffective.

The protein structure of HygR, as predicted by AlphaFold

Design

The transcriptional unit for the HygR marker (BBa_K4879009) is designed with the coding sequence flanked by the TEF1 promoter (BBa_K3629001) upstream and the XPR2 terminator (BBa_K3629004) downstream.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 153
  Illegal EcoRI site found at 247
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 153
  Illegal EcoRI site found at 247
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 153
  Illegal EcoRI site found at 247
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 153
  Illegal EcoRI site found at 247
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 153
  Illegal EcoRI site found at 247
  
• 1000
  COMPATIBLE WITH RFC[1000]