Part:BBa_K4482012:Design

HIRMAb-HC-NT3

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1917
  Illegal EcoRI site found at 2010
  Illegal XbaI site found at 1771
  Illegal SpeI site found at 262
  Illegal SpeI site found at 2026
  Illegal PstI site found at 1354
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1917
  Illegal EcoRI site found at 2010
  Illegal SpeI site found at 262
  Illegal SpeI site found at 2026
  Illegal PstI site found at 1354
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1917
  Illegal EcoRI site found at 2010
  Illegal BamHI site found at 1415
  Illegal XhoI site found at 16
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1917
  Illegal EcoRI site found at 2010
  Illegal XbaI site found at 1771
  Illegal SpeI site found at 262
  Illegal SpeI site found at 2026
  Illegal PstI site found at 1354
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1917
  Illegal EcoRI site found at 2010
  Illegal XbaI site found at 1771
  Illegal SpeI site found at 262
  Illegal SpeI site found at 2026
  Illegal PstI site found at 1354
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1077
  Illegal BsaI.rc site found at 39
  Illegal SapI.rc site found at 387
  

Design Notes

The sequences for HIRMAb-HC and NT-3 were ordered from Twist Bioscience as gene fragments and combined together with Gibson Assembly into a mammalian expression vector.

Source

The coding sequence for NT-3 was obtained from NCBI database (NP_001096124) and it was codon optimized for its synthesis by Twist Bioscience. The protein sequence for HIRMAb-HC was obtained from US patent US-8858950-B2 and back translated to obtain a DNA coding sequence for the protein, this DNA coding sequence was codon optimized for synthesis by Twist Bioscience and for expression in mammalian cells.