Part:BBa_K2450201
TetR with TEV cleavage site and CFP tag
This part is a modified TetR repressor protein containing a cleavage site for TEV protease. It offers a new way of relieving repression of a DNA system. The cleavage site is located in the linker between the DNA binding domain and dimerisation domain of TetR. It also has a His-tag for easy purification in a nickel column. It is an improvement of a previous iGEM part.
The TetR repressor is only functional as a dimer. Each monomer has a DNA-binding domain and a dimerisation domain.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
TetR functions as a homodimer to bind tet operator sequences and prevent the association of RNA polymerase with its promoter. Each TetR monomer has a DNA binding domain and a dimerisation domain. The DNA binding domain is a helix-turn-helix motif that associates with a tet operator sequence (TCCCTATCAGTGATAGAGA). Our complementary part, BBa_K2450301, contains 2 repeats of this sequence - one for each monomer.
TEV protease has a specific cleavage site of Glu-Asn-Leu-Tyr-Phe-Gln-(CUT)-Gly, which was inserted into our protein in a linker region between the DNA binding domain and the dimerisation domain.
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