Part:BBa_K4578003

INP-OspA

The ice nucleation protein (INP) allows for OspA to be expressed on the surface of E. coli, which is useful for future tests and diagnostics in order to determine if we can successfully detect B. burgdorferi.

Optimizing this sequence for E. coli allows for the design of a model organism that satisfies BSL-1 laboratory requirements. This provides more opportunities to test novel Lyme disease detection systems in the future.

OspA is expressed on the surface of B. burgdorferi and functions as a signal transducer or as a receptor (Li, 1997). The OspA sequence used BBa_K3767004 from iGEM21_Queens_Canada. The ice-nucleating protein (INP) is a natural surface expressing protein, and we hypothesized that by linking OspA to the INP, OspA would be expressed on the surface of the cell. The INP sequence was provided by Air Force Research Laboratory.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 849
  Illegal EcoRI site found at 1416
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 849
  Illegal EcoRI site found at 1416
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 849
  Illegal EcoRI site found at 1416
  Illegal BglII site found at 1467
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 849
  Illegal EcoRI site found at 1416
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 849
  Illegal EcoRI site found at 1416
  Illegal NgoMIV site found at 551
  Illegal AgeI site found at 405
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 580
  Illegal SapI.rc site found at 238