Part:BBa_K2520006

TRE-GFP-hGH

This device allows inducible expression of GFP within mammalian cells. The TRE promoter is made up of Tet operator (TetO) sequence concatemers fused to a minimal hCMV. This promoter can be used for inducible expression of the downstream gene in two different schemes.

Tet-off

In the absence of Dox, tTA binds to the TRE promoter and activates transcription of the downstream gene. In the presence of Dox, tTA cannot bind to the TRE, and expression from the target gene remains minimal.

Tet-on

In the presence of Dox, rtTA binds the TRE promoter and activates transcription of the downstream gene. In the absence of Dox, rtTA cannot bind the TRE promoter, and expression of the downstream gene is minimal.

Usage and Biology

In this device, the inducible protein is GFP, and can be used easily for calibration and characterization of inducible systems. The human growth hormone (hGH) terminator has been added to allow for efficient expression of the GFP.

Figure 1: Weighted median of cells expressing GFP depending on different concentrations of Doxycyline

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 site found at 1438
  Illegal BsaI.rc site found at 967