Part:BBa_K1061013

P tight promoter

It is a response element containing 7 consecutive TetO elements and a minimal CMV promoter. When binding to tTA or rtTA, the expression of the downstream gene will be highly activated. Besides, it can achieve low constitutive expression(almost undectectable) when not binding to tTA or rtTA.

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]

Funtional experiment

We conducted the functional experiment by co-tranfected the plasmid contains tTA and the plasmid contains P tight into Bosc, one kind of HEK-293 cell lines.Noted that we did not use tTA advance due to time limit(we construct the plasmid contains tTA advance later), and we will try that later,for improvement of expression control.

functional assay of tTA

The after adding dox, the expression of dox is almost fully suppressed. We also get the quantitive data by western blot, which will be submitted later.

Measurement Contribution

• Group: Tsinghua-A, 2016
• Author: Yuxi Ke
• Summary: We quantitatively measured and calculated TRE promotor's ability to accurately transmit information by channel capacity, a concept in information theory, the unit of which is bit.

• Link:http://2016.igem.org/Team:Tsinghua-A/Description

• Our Improvement
  • Purpose:Channel capacity evaluates correlation between input and output signals. For instance, if channel capacity is larger than 1 bit, the part can decern between high and low inputs, and vice versa. By assessing this novel quantity, we through light on how much information can a inducible promotor pass on. The measuring and calcualtion methods can be applied to more promotors in the future.
  • Results:Using EBFP2 as reporter, we measure channel capacity of a simple circuit driven by TRE to be 0.5259, with standard deviation of 0.0058.

channel capacity of TRE

• • Methods
• Uploads: (links to uploads relevant to your contribution, ex: csv containing your data, sequence files, etc.) Note: All files pertaining to a contribution must be uploaded to the Registry.