Part:BBa_K4161508
488aa recombinant protein, Aga2 + ipaD
This part is used to express a fused protein containing the ipaD antigen. contains a GAL1 promoter that allows the sequence being highly expressed in S. cerevisiae and a T7 promoter that plays a role in plasmid cloning in E. coli. Aga2 gene and ipaD gene express Aga2 proteins and ipaD antigenas, respectively. A (G4S)3 polypeptide sequence between the two genes serves as a linker sequence. The 3' end of the ipaD gene (or the C-terminus of the ipaD protein), is connected to a HA tag that enable the expressed fused protein to be purified via anti-HA antibody.
Usage and Biology
Usage
We expect this sequence to have the ability to express a fused protein containing ipaD antigen on the surface of S. cerevisiae. Therefore, the ipaD gene can be either fused with a signaling sequence targeting the plasmic membrane or a protein that natrually exists on the membrane. We also expect the expressed protein be able to be purified so that we can do further testing. In order to prevent the protein-protein interaction from disrupting the normal function of Aga2 and ipaD or disturbing the binding between HA-anti HA antibodies, we add linker sequences between these proteins.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 935
Illegal PstI site found at 800
Illegal PstI site found at 940 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 935
Illegal NheI site found at 857
Illegal PstI site found at 800
Illegal PstI site found at 940
Illegal NotI site found at 961 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 935
Illegal BamHI site found at 920
Illegal XhoI site found at 968 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 935
Illegal PstI site found at 800
Illegal PstI site found at 940 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 935
Illegal PstI site found at 800
Illegal PstI site found at 940
Illegal AgeI site found at 70 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1681
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