Difference between revisions of "Part:BBa K838001"
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Any promoters already present must be removed since we want only want expression drive by LovTAP-VP16 and not constitutive expression. The part must have a polyA tail at the end as well.  
 
Any promoters already present must be removed since we want only want expression drive by LovTAP-VP16 and not constitutive expression. The part must have a polyA tail at the end as well.  
  
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[[File:Team_EPF_Lausanne_simpleswitch.png|500px]]
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[[File:Team_EPF_Lausanne_simpleswitch.png]]
  
  

Revision as of 17:43, 26 September 2012

LovTAP readout This part needs to be used with part BBa_K838000, which is LovTAP-VP16.

The part consists of a TRP repressor and DsRed. The LovTAP-Vp16 protein attaches to the this readout and promotes transcription of the DsRed mRNA.


1) Cloning:

First clone the part into a mammalian expression vector such as [http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product pcDNA3.1(+)] or [http://products.invitrogen.com/ivgn/product/V04450 pCEP4]. These are the two main expression vectors we used.

Any promoters already present must be removed since we want only want expression drive by LovTAP-VP16 and not constitutive expression. The part must have a polyA tail at the end as well.

File:Team EPF Lausanne simpleswitch.png


2) Transfection Co-transfect the combination of LovTAP-Vp16 and readout in mammalian cells!


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 424
    Illegal SpeI site found at 295
    Illegal SpeI site found at 303
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 424
    Illegal SpeI site found at 295
    Illegal SpeI site found at 303
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 424
    Illegal BglII site found at 381
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 424
    Illegal SpeI site found at 295
    Illegal SpeI site found at 303
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 424
    Illegal SpeI site found at 295
    Illegal SpeI site found at 303
    Illegal NgoMIV site found at 7
  • 1000
    COMPATIBLE WITH RFC[1000]