Generator

Part:BBa_K779602

Designed by: Giulio Alighieri   Group: iGEM12_MIT   (2012-09-29)

Hef1a-TagBFP-FF5 MammoBlock Device

A composite part with a constitutive Hef1A promoter with a TagBFP (blue fluorescent protein) appended with microRNA binding sites. Created through the gateway cloning protocol.

Tagchar.png
We transfected 250 ng of Hef1A:TagBFP-FF5 into 100,000 HEK293 cells using Lipofectamine 2000 reagent. Images were taken on a Leica Confocal microscope at 10X. Left shows a brightfield image. Right shows the blue channel. We can observe high transfection efficiency and expression of TagBFP.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 315
    Illegal PstI site found at 820
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal NotI site found at 1285
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 569
    Illegal XhoI site found at 968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 315
    Illegal PstI site found at 820
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal NgoMIV site found at 703
    Illegal AgeI site found at 81
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1918


[edit]
Categories
Parameters
n/aHef1a-TagBFP-FF5 MammoBlock Device