DNA

Part:BBa_K1732004

Designed by: Donna Lee   Group: iGEM15_Carnegie_Mellon   (2015-09-07)
Revision as of 03:38, 8 September 2015 by Donnalee (Talk | contribs)

pSB1C3-J23100-B0034-CDcel-Gaussia-His-B0015

J23100 constitutive promoter (BBa_J23100), a RBS (BBa_B0034), CDcel domain (BBa_K1732002), Gaussia luciferase (BBa_K1732003), and B0015 terminator (BBa_B0015). ______________________________________________________________________________________________________________________________

CDcel Gaussia.jpg


This DNA sequence, expressed from a constitutive, strong promoter allows for the Gaussia luciferase protein to be synthesized and was targeted extrecellularly into the media. The Gaussia luciferase proteins were reacted with the optimal concentration of coelenterazine to express luminescence, which was used to test the Carnegie Mellon DIY luminometer. The sequence was codon optimized for E.coli and contains a 6X His-tag for easier purification of the protein.


CDcel Gaussia coleneterazine.jpg


Kinetics of luciferase.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1517
    Illegal BamHI site found at 975
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 715


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Categories
Parameters
None