Part:BBa_K5371201:Design

pAbAi-GAL4-NH

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 7
  Illegal EcoRI site found at 3600
  Illegal XbaI site found at 917
  Illegal PstI site found at 1560
  Illegal PstI site found at 4214
  Illegal PstI site found at 7688
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 7
  Illegal EcoRI site found at 3600
  Illegal PstI site found at 1560
  Illegal PstI site found at 4214
  Illegal PstI site found at 7688
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 7
  Illegal EcoRI site found at 3600
  Illegal BamHI site found at 48
  Illegal BamHI site found at 3331
  Illegal XhoI site found at 429
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 7
  Illegal EcoRI site found at 3600
  Illegal XbaI site found at 917
  Illegal PstI site found at 1560
  Illegal PstI site found at 4214
  Illegal PstI site found at 7688
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 7
  Illegal EcoRI site found at 3600
  Illegal XbaI site found at 917
  Illegal PstI site found at 1560
  Illegal PstI site found at 4214
  Illegal PstI site found at 7688
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 348
  Illegal BsaI.rc site found at 2024
  Illegal BsaI.rc site found at 3213
  Illegal BsaI.rc site found at 5939
  Illegal BsaI.rc site found at 6998
  Illegal SapI site found at 713
  Illegal SapI site found at 4856
  Illegal SapI site found at 7150
  

Design Notes

Stop codon of mCherry homologous region was removed for continuous and complete expression of Sir2-linker-mcherry to prevent a premature stop in this case. Length of the linker is adjusted to avoid frameshift, which may cause faulty translation. Codons of the linker is designed to avoid polarized amino acids or those with large residues.

Source

GAL4_NH1574_V2 was constructed via Gibson Assembly, with segments mainly amplified from Saccharomyces cerevisiae.