Part:BBa_K5301028:Design
spMSP1D1 expression pathway (T7)
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 571
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 571
Illegal NheI site found at 139
Illegal NheI site found at 1145 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 571
Illegal BglII site found at 999
Illegal BamHI site found at 172
Illegal XhoI site found at 895 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 571
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 571
- 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.MSP1D1 (Membrane Scaffold Protein 1D1) is a synthetic protein that is a derivative of apolipoprotein A-I, the main protein component of high-density lipoprotein (HDL). MSP1D1 can self assemble and form disc-shaped nanoparticles in the presence of synthetic phospholipids, known as' nanodiscs'.