Difference between revisions of "Part:BBa K2100003"
 
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__NOTOC__
 
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<partinfo>BBa_K2100003 short</partinfo>
 
<partinfo>BBa_K2100003 short</partinfo>
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The pENTR hEF1a part is an entry vector containing the human elongation factor 1 alpha (hEF1a) promoter, a constituitive mammalian promoter.
  
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Enter description about hef1a promoter here.
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We have characterized the hEF1a promoter in all four cell lines we experimented on in the duration of our project using the following fluorescent tags: eYFP, mKate, and tagBFP.  
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The pENTR hEF1a part is an entry vector containing the human elongation factor 1 alpha (hEF1a) promoter, acting as a constituitive mammalian promoter.
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We have characterized the hEF1a promoter in all four cell lines we experimented on in the duration of our project using the following fluorescent tags: eYFP, mKate, and (tag?)BFP.  
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
<center><img src= "https://static.igem.org/mediawiki/parts/a/ae/T--MIT--attractenehef1aeyfp.png" alt = '' style="width:800px;height:412px;"  margin: 0 1.5%; class="rotate90"></center>
 
<center><img src= "https://static.igem.org/mediawiki/parts/a/ae/T--MIT--attractenehef1aeyfp.png" alt = '' style="width:800px;height:412px;"  margin: 0 1.5%; class="rotate90"></center>
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Our team used the entry vector of hEF1a to create an expression vector with many constructs. We often used
  
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2100003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2100003 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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Latest revision as of 23:06, 28 October 2016


pENTR hEF1a

The pENTR hEF1a part is an entry vector containing the human elongation factor 1 alpha (hEF1a) promoter, a constituitive mammalian promoter.

We have characterized the hEF1a promoter in all four cell lines we experimented on in the duration of our project using the following fluorescent tags: eYFP, mKate, and tagBFP.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal BglII site found at 591
    Illegal BamHI site found at 1197
    Illegal XhoI site found at 990
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
    Illegal NgoMIV site found at 725
    Illegal AgeI site found at 103
  • 1000
    COMPATIBLE WITH RFC[1000]

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