Generator

Part:BBa_K332032:Design

Designed by: KUO, CHIA-CHUN   Group: iGEM10_NCTU_Formosa   (2010-10-22)

Constitutive promoter+37℃ induced RBS+tetR+double terminator+Ptet+RBS+GFP+double terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 794
    Illegal NgoMIV site found at 1723
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1639


Design Notes

To regulate GFP gene by temperature.

To maintain cells at 37° C ↑ : the tetR protein be generated and repress Ptet's circuit.

To release cells at 37° C ↓  : the tetR protein not be generated and do not repress Ptet's circuit.


TPcon1.jpg


The theoretical switching temperature of BBa_k115002, which was designed by Team TUDelft in 2008, is 37℃.

Thus, our thermosensing device A can be induced at this temperature. (The switching temperature can be modified by changing RBS according to different conditions.)

Once being swiched on, tetR will inhibit the corresponding promoter, pTet, which enables us to regulate the expression of cry gene and quorum sensing gene devices.



sequencing's blast:

A+B 1.jpg A+B 2.jpg A+B 3.jpg A+B 4.jpg

Source

the 37℃ induced RBS is from iGEM08_TUDelft

References