Composite

Part:BBa_K191003

Designed by: Le Thanh Tu NGUYEN   Group: iGEM09_EPF-Lausanne   (2009-10-11)

Promoter Lac I - RBS - LovTAP - Term

Part BBa_K191003 codes for the hybrid protein LovTAP. This hybrid protein has a light-sensitive domain (Lov means : light, oxygen, voltage) and a regulatory domain (part of the Trp repressor). The fused protein allows transmission of the conformational change induced by light (on the light-sensitive domain) to the DNA-binding domain, therfore resulting in an increase or decrease of the regulatory domain's affinity for its DNA binding site. The activation of the LovTAP protein (binding to DNA) is achieved by illumination @ 470 nm

For more details, please consult the section of [http://2009.igem.org/Team:EPF-Lausanne/LOVTAP LovTAP system ] on team EPF-Lausanne wiki.

Usage and Biology

In order to be able to test the efficiency of the fused protein LovTAP, its expression is induced by IPTG (thanks to the LacI promoter). Two other parts were also constructed as the read out systems of LovTAP : BBa_K191004 and BBa_K191005

All three parts together give the following system, where LovTAP activities can be measured through RFP fluorescence intensity (both with with static and kinetic measurements)

Systems.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 764


[edit]
Categories
//function/sensor/light
Parameters
input_sIPTG
positive_regulators
protein_outLovTAP