Regulatory

Part:BBa_K079038

Designed by: Francesca Ceroni   Group: iGEM08_Bologna   (2008-10-25)

TetO-wt/4C5G - operator library member

These parts have been extracted from the TeT Operator Library.

Repression efficiency of Tet Operators Variants expressed as β-galactosidase percentage activity. 100% Being unrepressed

This subset of operator sequences has been chosen since it allows to cover a wide affinity and repression range


K079036:Tet O is the Wild Type operator sequence regulating Tetracycline resistance genes. The natural sequence exhibit a tigth repression given by an high Tet R bindig affinity
K079037:TetO-4C is a medium affinity operator obtained through a T-C mutation in the fourth nucleotide on both side of the symmetric consensus sequence. Disrupting this interaction the repression efficiency is about 12 time lower than the Wt
K079038: TetO-wt/4C5G is a double mutant that heavily compromises the interaction of TetR with the right half of the binding sequence. The resulting repression is quite weak, more than 40 times less stringent than in TetO


The parts can be individually assembled with Constitutive Promoter Family yielding a collection of Tc inducible promoters with different dynamics. Design rules and effect of multiple Tet Operator sites on repression strength and cooperativity are scarcely represented in the literature. Further investigation in this direction are planned

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]


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Parameters
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