Coding

Part:BBa_K2450201:Design

Designed by: Zoe Ford   Group: iGEM17_Oxford   (2017-09-27)


TetR with TEV cleavage site and CFP tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We had to decide on an appropriate location for the TEV cleavage site in TetR. We examined the crystal structure of TetR and identified a linker between the DNA binding and dimerization domains. We decided to have a C-terminal CFP tag so it would not interfere with the N-terminal DNA binding domain of the TetR. We used CFP so it would be compatible with our other parts, using mCherry and eYFP, and all would be visible in our fluorescent microscope as they have distinct absorption and emission wavelengths.

Source

TetR from BBa_K106669

TEV cleavage sequence from Merck

CFP from AddGene

References

PDB entry 3zqi for structure of TetR