Difference between revisions of "Part:BBa K1150026"

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! colspan="2" style="background:#FFBF00;"|uniCAS red Light Switch Part II - Activator
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! colspan="2" style="background:#FFBF00;"|CMV:PhyB-VP16:BGH
 
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|'''Function'''
 
|'''Function'''

Revision as of 12:38, 4 October 2013

uniCAS Red Light Switch Part II - Activator


CMV:PhyB-VP16:BGH
Function Activation domain of red light

induced gene expression control

Use in Mammalian cells
RFC standard RFC 25
Backbone pSB1C3
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

Usage and Biology

Fusion protein of PhyB and VP16. This fusion protein PhyB-VP16 is an interaction partner of dCas9-PIF. When crRNA and tracrRNA bind to Cas9, the protein is able to bind complementary DNA. When the system is exposed to red light (660 nm), the Phytochrome B receptor binds to the Phytochrome interaction actor (PIF). So it recruits the dCas9-PIF protein. The PhyB-PIF binding can be abolished by illumination with far-red light (740nm.) This system enables activation of gene expression induced by red light. Have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 576
    Illegal BglII site found at 1076
    Illegal BamHI site found at 1158
    Illegal XhoI site found at 1109
    Illegal XhoI site found at 1128
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1325


References

Müller, K. et al. (2013). A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells. Nucleid Acid Research
Hirai, H. et al. (2010). Structure and functions of powerful transactivators: VP16, MyoD and FoxA. Int. J. Dev. Biol.