Part:BBa_K1172401:Design

mtrCAB - gene cluster from S. oneidensis for cytochrome expression

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2049
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 235
  

Design Notes

• Two illegal PstI restriction sites, which had to be removed by silent mutation via specific primers thus generating three fragments
  • pSB1C3_fwd:
  • pSB1C3_rev:
  • Fragment1_fwd:
  • Fragment1_rev:
  • Fragment2_fwd:
  • Fragment2_rev:
  • Fragment3_fwd:
  • Fragment3_rev:

• Fragments and backbone joined by Gibson assembly

Source

• Shewanella oneidensis MR-1 genomic sequence
  • The strain was kindly provided by Dr. Johannes Gescher (Karlsruher Institut für Technologie)