Part:BBa_K838001

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.

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]