DNA

Part:BBa_K1732000:Design

Designed by: Donna Lee   Group: iGEM15_Carnegie_Mellon   (2015-09-03)
Revision as of 08:11, 3 September 2015 by Donnalee (Talk | contribs) (→‎Design Notes)

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J23100-PelB-GlucHCO-B0015


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 546
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 115
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This sequence is codon optimized for E.coli.

The plasmid was used to produce Gaussia princeps luciferase proteins that would express luminescence when reacted with an optimal concentration of 3-4 uM of coelenterazine.

Gaussia Seq.jpg

Sequence contains a PelB leader sequence which suggested that the Gaussia princeps luciferase proteins would be secreted into the expected location of the periplasmic space. Howeverm the observed location of the proteins was extracellular (in the media).

Gaussia luminescence decay.jpg

Source

Engineered from XXXXXXXX. Provided by the Bruchez lab at Carnegie Mellon.

References