Difference between revisions of "Part:BBa K368001"

 
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CMV promoter induceable by Doxycycline. Only works in special Tet-On cell lines (stable transfected with pTet-On-Advanced Vector by clontech)!  
 
CMV promoter induceable by Doxycycline. Only works in special Tet-On cell lines (stable transfected with pTet-On-Advanced Vector by clontech)!  
 
Also works in special Tet-Off cell lines (stable transfected with pTet-Off-Advanced Vector by clontech), but then the promoter will be inactivated by Doxycycline.
 
Also works in special Tet-Off cell lines (stable transfected with pTet-Off-Advanced Vector by clontech), but then the promoter will be inactivated by Doxycycline.
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===Improvement by SYSU-CHINA 2018===
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In iGEM 2018, SYSU-CHINA improved the part by submitting a almost identical promoter sequence [[BBa_K2748002]] used in their project for teams in the future. They also characterized the promoter function using both fluorescence imaging and western blot, obtaining results on optimal induction concentration and the time course of expression, which may benefit teams in the future. In addition, a coding sequence of reverse tet-responsive transactivator [[BBa_K2748001]] was submitted to the registry, so that teams in the future can establish their own stable cell lines for for tet-inducible expression, or integrate it into vectors containing tet-ON promoter (like we did!) for tet-inducible expression of gene of interest.
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Feel free to check out our new basic part [[BBa_K2748002]]!
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 19:35, 17 October 2018


minimal CMV Promoter with TRE (tet responsible element)

CMV promoter induceable by Doxycycline. Only works in special Tet-On cell lines (stable transfected with pTet-On-Advanced Vector by clontech)! Also works in special Tet-Off cell lines (stable transfected with pTet-Off-Advanced Vector by clontech), but then the promoter will be inactivated by Doxycycline.


Improvement by SYSU-CHINA 2018

In iGEM 2018, SYSU-CHINA improved the part by submitting a almost identical promoter sequence BBa_K2748002 used in their project for teams in the future. They also characterized the promoter function using both fluorescence imaging and western blot, obtaining results on optimal induction concentration and the time course of expression, which may benefit teams in the future. In addition, a coding sequence of reverse tet-responsive transactivator BBa_K2748001 was submitted to the registry, so that teams in the future can establish their own stable cell lines for for tet-inducible expression, or integrate it into vectors containing tet-ON promoter (like we did!) for tet-inducible expression of gene of interest.

Feel free to check out our new basic part BBa_K2748002!

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]