Part:BBa_I759017:Design

cis5-repressed, tet-regulated YFP

Assembly Compatibility:

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

  Illegal NheI site found at 985
  Illegal NheI site found at 1008
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

This part is the YFP sequence down-regulated by the cis repressive element. The Ptet promoter and TetR gene, which is under the control of a constitutive promoter, establish an inducible system in which transcription can be activated by the addition of anhydrotetracycline (aTc).

Cis5 is a modified version of constructs made by the University of California Berkeley iGEM 2006 team. Modifications involve a change in the number of bulges and end-loop size which affect the free energy of interaction within the cis structure. The key principal in the design was to find a structure that offered tight repression of the gene of interest but also allowed for activation upon addition of a trans-activating element. A lower free energy indicates tighter binding and a decreased favorability for forming a compliment with the trans element. The cis5 construct was designed to be activated by trans3 and trans 5. Design specifics: free energy = -12.6kcal; 6bp loop, 4 bulges, 5bp toe-hold

Source

Caltech iGEM 2007. The cr element is a synthetic oligo which has been inserted into part I759009.

References

http://parts2.mit.edu/wiki/index.php/University_of_California_Berkeley_2006