Difference between revisions of "Part:BBa K2100006"
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2100006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2100006 SequenceAndFeatures</partinfo>
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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.
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https://static.igem.org/mediawiki/parts/c/c8/T--MIT--hEF1a.png
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Here is an image of tHESC cells constitutively expressing mKate florescence under the hEF1a promoter.
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https://static.igem.org/mediawiki/parts/9/92/T--MIT--khb_thesc_red.jpeg
  
  

Latest revision as of 23:07, 28 October 2016


pENTR mKate

mKate is a red fluorescent protein.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 647
    Illegal SapI.rc site found at 29

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 tHESC cells constitutively expressing mKate florescence under the hEF1a promoter.

T--MIT--khb_thesc_red.jpeg