Generator

Part:BBa_K511815

Designed by: Grant Robinson   Group: iGEM11_MIT   (2011-09-28)

Inducible Red Fluorescent Protein Generator (TRE-Tight-mKate) MammoBlock Device

This MammoBlock composite device produces the monomeric red fluorescent protein mKate when induced with tTA/rtTA transactivator variants in the presence of tetracycline analogues, and otherwise produces mKate at a low, basal (OFF) level.

Here is a graph of the activation of TRE_mKate with constitutively expressed rtTA3 over a range of doxycycline concentrations.

Tre_mkate.jpg

Figures 1, 2, and 3 below show FACS expression data for HEK-293 cells without any construct, HEK-293 cells with this construct and zero doxycycline, and HEK-293 cells with this construct and 2 ug/mL of doxycycline, respectively. Additionally, a Hef1a-rtTA3 plasmid was used in all cases to provide the transactivator necessary for TRE-mediated expression.





Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal XbaI site found at 30
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal XbaI site found at 30
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal XbaI site found at 30
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 986
    Illegal SapI.rc site found at 368


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Categories
Parameters
n/aInducible Red Fluorescent Protein Generator (TRE-Tight-mKate) MammoBlock Device