Part:BBa_K4294780

TetR TU

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This is the transcriptional unit for TetR repressor under a constitutive promoter (BBa_J23106 ) to regulate the pTet promoter.

Circuit Design

TetR forms a dimer and binds to tet operators in PTet promoter and blocks the assembly of the transcription machinery. PTet is a medium strength promoter that is constitutively “ON” in the absence of TetR. It contains two tet operators, one inside the core promoter sequence and the other after the -10 hexamer, leading to efficient repression. The system is efficiently induced via anhydrotetracycline hydrochloride (aTc), a tetracycline derivative which binds Tet R with an ~35-fold higher binding constant, being an effective inducer at very low concentrations. Additionally, its antibiotic activity is ~100-fold lower and has minimal toxic effect on E.coli cells in the concentration used for induction.

Measurment

This TU was proven to not be functional for our system. We failed to induce our system with aTc using this promoter. We do not recommend it for use in similar circuits.

References

[1] Lutz, R., & Bujard, H. (1997). Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic acids research, 25(6), 1203–1210. https://doi.org/10.1093/nar/25.6.1203

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]