Measurement

Part:BBa_K772009

Designed by: Mustafa Elitok   Group: iGEM12_Fatih   (2012-09-26)

GFP - LasR protein complex with flag tags

This part contains a green fluorescent protein (GFP) as a non-reactive chemical compound (anchor) in order to neutralize the active reactive fixed to it, an inducer regulatory protein (LasR is responsible to induce pLas promoter in the presence of 3OC12HSL which is a chemical compound sythesized by an enzyme that LasI gene codes.) and a TEV protease cleavage site between them to assemble these two proteins. BBa_772001[1] and BBa_K772002[2] are assembled to be used as the trigger of detection in the whole system which ends with the intracellular transfection (or activation) of TEV protease. After the activation, it is planned that TEV protease will cleave this anchor-inducer protein complex to release free the reporter. In conclusion, in this system, the inducer would activate a transcription pathway on the targeted inducible gene or the reporter would indicate that there is a change in the whole system.

This part contains flag tags extra from BBa_K772007[3] in the GFP region; because it is planned to be used in the Co-IP/SDS-PAGE assay to be isolated.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1180
    Illegal AgeI site found at 1377
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 693


[edit]
Categories
//classic/regulatory/other
//classic/reporter
//function/regulation/transcriptional
//function/reporter/fluorescence
//proteindomain/cleavage
//proteindomain/tail
Parameters
activation_coeficientTEV Protease
chassisE.coli TOP10 strain
device_typeQuorum Sensing
functionRegulatory
proteinsGFP, LasR
targetpLas