Difference between revisions of "Part:BBa K2100003"
| Line 17: | Line 17: | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2100003 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2100003 SequenceAndFeatures</partinfo> | ||
| + | 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. | ||
| + | |||
| + | https://static.igem.org/mediawiki/parts/c/c8/T--MIT--hEF1a.png | ||
| + | |||
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
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.
