Difference between revisions of "Part:BBa K2485012:Design"

(Design Notes)
(Source)
Line 15: Line 15:
 
The cPT Region of IcsA was sourced from BBa_K2485011 with removed start and stop codons, and the last four nucleotides were modified to match the fusion sequence between cPT and GFP in a plasmid provided by Marcia B. Goldberg, MD which was experimentally shown to unipolarly localize GFP.
 
The cPT Region of IcsA was sourced from BBa_K2485011 with removed start and stop codons, and the last four nucleotides were modified to match the fusion sequence between cPT and GFP in a plasmid provided by Marcia B. Goldberg, MD which was experimentally shown to unipolarly localize GFP.
  
FresnoRFP from Atum was used instead of GFP.
+
FresnoRFP from Atum was used to maximize the distance between the absorption/emission of the fluorescent protein (553/592) and the absorption of the proteorhodopsin (~520).
  
 
===References===
 
===References===

Revision as of 19:27, 21 September 2017


Proteorhodopsin-cPT-RFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1117
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

A 5x Gly-Ser linker, BBa_J18922, was used to join proteorhodopsin and cPT.

Source

The green light absorbing protein rhodopsin was sourced from BBa_K773002 and codons were slightly modified to enhance GC content. It is a proton pump used by marine bacteria for generating proton motive force for ATP generation.

The cPT Region of IcsA was sourced from BBa_K2485011 with removed start and stop codons, and the last four nucleotides were modified to match the fusion sequence between cPT and GFP in a plasmid provided by Marcia B. Goldberg, MD which was experimentally shown to unipolarly localize GFP.

FresnoRFP from Atum was used to maximize the distance between the absorption/emission of the fluorescent protein (553/592) and the absorption of the proteorhodopsin (~520).

References