Part:BBa_K4578008

INP-OspA expression block

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.

This expression block includes INP-OspA and its respective T7 promoter, lac operon, and RBS.

OspA is expressed on the surface of B. burgdorferi and functions as a signal transducer or as a receptor. 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 1038
Illegal EcoRI site found at 1605
Illegal XbaI site found at 47
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1038
Illegal EcoRI site found at 1605
Illegal NheI site found at 151
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1038
Illegal EcoRI site found at 1605
Illegal BglII site found at 1656
Illegal BamHI site found at 184
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1038
Illegal EcoRI site found at 1605
Illegal XbaI site found at 47
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1038
Illegal EcoRI site found at 1605
Illegal XbaI site found at 47
Illegal NgoMIV site found at 740
Illegal AgeI site found at 594
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 769
Illegal SapI.rc site found at 427