Part:BBa_K3861011

tetR-Ptet

Tetracycline inducer system based on the tetracycline resistance mechanism found in gram-negative bacteria.¹ It is one of the most used transcriptional regulatory systems.² We use the Tet-On system where the transcription is induced in the presence of a tetracycline or tetracycline derivates.³ The rtTA (reverse tetracycline transactivator) protein, a fusion of the TetR (tetracycline repressor) and VP16 ativation domain, can bind the TRE (tetracycline response element) only in the presence of tetracycline, thereby activating gene expression.³

Usage and Biology

The sequence contains already an RBS, thus genes that aimed to be expressed from this promoter can be added directly downstream of the submitted sequence. The sequence is derived from Tn10dTet transposon.⁴ The sequence upstream of tetR is necessary to prevent transcription into adjacent genes.

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]

References

1. Orth, P., Schnappinger, D., Hillen, W., Saenger, W. & Hinrichs, W. Structural basis of gene regulation by the tetracycline inducible Tet repressor–operator system. Nat. Struct. Biol. 7, 215–219 (2000).
2. Baron, U. & Bujard, H. Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Methods Enzymol. 327, 401–421 (2000).
3. Gossen, M. et al. Transcriptional Activation by Tetracyclines in Mammalian Cells. Science 268, 1766–1769 (1995).

4. Rappleye, C. A. and Roth, J. R. A Tn10 derivative (T-POP) for isolation of insertions with conditional (tetracycline-dependent) phenotypes. Journal of Bacteriology, 5827-5834 (1997).