Difference between revisions of "Part:BBa K1119008"
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iGEM Technion 2017 improved this part to <partinfo>BBa_K2520006</partinfo>. This improved device contains a TRE promoter instead of the original CMV, to allow for inducible expression of GFP within mammalian cells. | iGEM Technion 2017 improved this part to <partinfo>BBa_K2520006</partinfo>. This improved device contains a TRE promoter instead of the original CMV, to allow for inducible expression of GFP within mammalian cells. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Latest revision as of 17:07, 30 October 2017
CMV promoter - GFP - hGH polyA tail
This is a mammalian GFP generator used for characterization of our CMV promoter (BBa_K1119006). This GFP generator contains the CMV promoter sequence with GFP reporter in RFC25 format(BBa_K648013) and hGH polyA terminator (BBa_K404108).
This GFP generator was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope.
pEGFP-N1 (Clontech) served as the positive control. It contains the CMV promoter and EGFP reporter. A GFP generator that does not contain the CMV promoter was used as the negative control.
The [http://2013.igem.org/Team:Hong_Kong_HKUST/characterization detailed protocol] of our characterization can be found in HKUST iGEM 2013 Wiki.
Improved Part
iGEM Technion 2017 improved this part to BBa_K2520006. This improved device contains a TRE promoter instead of the original CMV, to allow for inducible expression of GFP within mammalian cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 614
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 628
Illegal AgeI site found at 1369 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1772
Illegal BsaI.rc site found at 1274