Part:BBa_K3861027

tetR-Ptet-GFP(STm codon harmonized)

We have cloned our Salmonella codon harmonized GFP to a tetracycline-controlled transcriptional activation system (pTet-tetR) (BBa_K3861027{https://parts.igem.org/wiki/index.php?title=Part:BBa_K3861027}; [BBa_K3861011 + BBa_K3861019]). The tetR system is well characterized and may be the most used gene expression controlling promoter system to date.1 There are two variants of this promoter-induction system, called Tet-On and Tet-Off.2,3 We use the Tet-On variant that selectively activates expression in the presence of tetracyclines. Important for our needs is the fact that tetracycline (or Anhydro-tetracycline) can passively penetrate bacterial membranes and no active transportes systems is crucial for this inducible expression system.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1382
  

Reference

1. Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. U. S. A. 89, 5547–5551 (1992). 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. Berens, C. & Hillen, W. Gene regulation by tetracyclines. Constraints of resistance regulation in bacteria shape TetR for application in eukaryotes. Eur. J. Biochem. 270, 3109–3121 (2003).