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 | + | 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1117
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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).