DNA

Part:BBa_K2100005

Designed by: Kathleen Brandes   Group: iGEM16_MIT   (2016-10-10)


pENTR eYFP

eYFP is a yellow fluorescent protein.

Sequence and Features

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 222
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 222
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 222
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 222
    Illegal AgeI site found at 5
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal PstI site found at 222

The MIT iGEM team used the hEF1a promoter to constitutively express fluorescent proteins as transfection markers in our experiments in HEK293, tHESC, MCF-7, and ISH cells. The amount of fluorescent units observed is an indication of the number of plasmids a particular cell uptakes. The graph below shows two single color controls from one of our experiments. We transfected one well of HEK293 cells with hEF1a-mKate and another with hEF1a-eYFP. The cells that uptook the hEF1a-mKate plasmids showed an increase in only the red fluorescent output, while the cells that uptook the hEF1a-eYFP plasmids showed a clear increase in only the yellow fluorescent output.

T--MIT--hEF1a.png

Here is an image of HEK293 cells transfected with the eYFP florescent gene cascaded with a hEF1a constituitive promoter. We often used yellow florescence as either a reporter for how our promoter responded to induction in cells or as transfection markers for experiments.

796px-T--MIT--khb_eYFPexpression_hek.jpeg


[edit]
Categories
//awards/part_collection/2016
Parameters
None