Project
pGSH

Part:BBa_M36091:Design

Designed by: Jonathan Deaton   Group: Stanford BIOE44 - S11   (2014-10-23)


HeLa Cell Glutathione Sensor


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 246
    Illegal SpeI site found at 2175
    Illegal PstI site found at 1503
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2236
    Illegal SpeI site found at 246
    Illegal SpeI site found at 2175
    Illegal PstI site found at 1503
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 246
    Illegal SpeI site found at 2175
    Illegal PstI site found at 1503
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 246
    Illegal SpeI site found at 2175
    Illegal PstI site found at 1503
    Illegal AgeI site found at 111
    Illegal AgeI site found at 708
    Illegal AgeI site found at 1593
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The main restriction in this construct was keeping DNA length under 2500 base pairs. This required removal of a Thyroglobulin (TG) Enhancer sequence 1902 to 1744 base pairs upstream of TG transcription Initiation. This size limit also required us to eliminate the Transcription Termination Factor-1 (TTF-1), another transcription factor important in the activation of the TG promoter.


Source

http://www.journal-of-hepatology.eu/article/S0168-8278(01)00037-X/pdf http://www.thyroidmanager.org/chapter/ontogeny-anatomy-metabolism-and-physiology-of-the-thyroid/

References