Part:BBa_K5033006:Design
OncoBiotica: mFadA[B]_GSLinker_eGFP
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 115
Illegal XhoI site found at 835
Illegal XhoI site found at 842 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
When designing this part we designed primers for amplification of the eGFP insert. Find them in our materials list on our Experiments page.
Source
As discussed on the main page, the first protein domain is derived from the part BBa_K4990002 but it has been codon optimized for expression in E. coli. It is the mFadA B-domain, found in various Fusobacterium strains. This domain should be able to bind to FadA pili on Fusobacterium nucleatum and its former subspecies Fusobacterium nucleatum, F. polymorphum, F. vincentii, F. animalis via self assembly.
The second functional protein domain is linked to the mFadA B-domain by a synthetic flexible linker consisting of Glycin and Serine in alternating order. This linker is 12 amino acids long.
This second functional domain of the fusionprotein is eGFP which originally is native to Aequorea victoria (water jellyfish). The eGFP sequence was aqcuired using PCR methods on a template provided by the lab our team was working in. The fusionprotein encoded by this part also contains a downstream hexa-histidine tag for protein purification.