Difference between revisions of "Part:BBa K4497014"
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<partinfo>BBa_K4497014 short</partinfo> | <partinfo>BBa_K4497014 short</partinfo> | ||
| − | This part encodes the | + | This part encodes the sequence for a mCherry-nanobody based receptor for secretion/ surface expression. The receptor contains a Flag- and HA-tag for immunofluorescence labeling. |
==Design & Cloning== | ==Design & Cloning== | ||
Latest revision as of 14:04, 13 October 2022
mCherry Nanobody Receptor
This part encodes the sequence for a mCherry-nanobody based receptor for secretion/ surface expression. The receptor contains a Flag- and HA-tag for immunofluorescence labeling.
Design & Cloning
Design
This part is made of the following components:
- mCherry nanobody receptor (Part:BBa_K4497014)
- IL-11 Signaling Sequence (Part:BBa_K4497003): Signaling Sequence for Cell surface expression
- Flag-tag (Part:BBa_K4497012): Flag epitope tag that enables surface expression detection using immunofluorescence labeling
- HA-tag (Part:BBa_K1150016): HA epitope tag that enables surface expression detection using immunofluorescence labeling
- Anti-mCherry nanobody (Part:BBa_K4497002): a nanobody capable of binding mCherry
Cloning
The part is encoded on the plasmid pcDNA3.1-SP-Flag HA-Tag-mCherrynb-gp130 which was kindly provided to us by Prof. Scheller [1]
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 373
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 373
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 94
Illegal BamHI site found at 127 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 373
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 373
- 1000COMPATIBLE WITH RFC[1000]
References
[1] Mossner, S., Phan, H. T., Triller, S., Moll, J. M., Conrad, U., & Scheller, J. (2020). Multimerization strategies for efficient production and purification of highly active synthetic cytokine receptor ligands. PLOS ONE, 15(4), e0230804.