Difference between revisions of "Part:BBa K2450201"
(→‎Usage and Biology)
Line 13: Line 13:
  
 
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.
 
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.
 +
 +
[File:Oxford_2017_modified_TetR_full.png]
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 16:23, 21 October 2017


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


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]

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.

[File:Oxford_2017_modified_TetR_full.png]