Part:BBa_K5301029:Design
spNW15 expression pathway (T7)
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 571
Illegal SpeI site found at 523
Illegal PstI site found at 739 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 571
Illegal NheI site found at 139
Illegal NheI site found at 1175
Illegal SpeI site found at 523
Illegal PstI site found at 739 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 571
Illegal BglII site found at 627
Illegal BglII site found at 999
Illegal BamHI site found at 172 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 571
Illegal SpeI site found at 523
Illegal PstI site found at 739 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 571
Illegal SpeI site found at 523
Illegal PstI site found at 739
Illegal AgeI site found at 1102 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We need to consider the strength of the promoter, the issue of achieving self circularization of MSP, and the subsequent protein purification.
Source
The SpyCatcher-SpyTag system is a protein binding method based on a modified domain of the SpyCatcher surface protein of Streptococcus pyogenes, which can recognize a homologous 13 amino acid peptide (SpyTag). After recognition, covalent heteropeptide bonds will form between the lysine side chain in SpyCatcher and the aspartic side chain in SpyTag.The NW15 protein is obtained through a specific truncation process from the NW30 protein. The NW30 protein itself is a protein synthesized through genetic engineering technology, which is modified from a portion of human apolipoprotein A-I. The NW30 protein can self assemble and form disc-shaped nanoparticles in the presence of synthesized phospholipids, known as' nanodiscs'.