Difference between revisions of "Part:BBa K511815"

 
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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.
 
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.
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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.
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<html><img src='https://static.igem.org/mediawiki/parts/e/e0/TREMKATEBLANK.jpg
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' style="width:30%"><br></html>
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<html><img src='https://static.igem.org/mediawiki/parts/e/e1/TREMKATE0.jpg
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' style="width:30%"><br></html>
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<html><img src='https://static.igem.org/mediawiki/parts/d/da/TREMKATEMAX.jpg
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Revision as of 05:07, 29 September 2011

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.

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