Part:BBa_K1761001
Outer Membrane Protein X (OmpX) with BamHI-linker and mNeonGreen
This part consists of three subparts, namely the uter membrane protein OmpX, a BamHI linker and the fluorophore mNeonGreen. OmpX is an outer membrane protein with the C- and N-termini in the intracellulair domain. To be able to use OmpX as a scaffold, a non-natural amino acid needs to be introduced. This can be done by implementing the amber stop codon TAG in one of the loops of OmpX via a mutation. With a specific tRNA an azide-functionalized amino acid can be built in, which can be used for the SPAAC click chemistry reaction with DBCO functionalized groups. The BamHI linker is a 213 bp long flexible GGSGGS linker. Using the restriction enzyme BamHI, the linker can become 45 bp shorter. mNeonGreen is a yellow-green fluorescent protein. For a fluorescene measurement, the mNeonGreen fluorophore can be excitated with a laser with a wavelenght of 480 nm and readed out at 517 nm.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 574
Illegal BamHI site found at 619 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
//function/fret
| emission | 517 nm |
| excitation | 480 nm |