Part:BBa_K4244046
flipUM (MMP-9 inducible)
"This flipUM is designed in such a way that it will only regain its colour when it is cleaved by MMP-9. Two inserts have been made, the E5 and K5 domain. These two domains dimerize and disrupt GFP structure. MMP-9 recognizes the cleavage site and cuts off the K5 domain, restoring Ultramarine function.
The flip design is based off the following paper: Zhang, Q., Schepis, A., Huang, H., Yang, J., Ma, W., Torra, J., Zhang, S. Q., Yang, L., Wu, H., Nonell, S., Dong, Z., Kornberg, T. B., Coughlin, S. R., & Shu, X. (2019). Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals. Journal of the American Chemical Society, 141(11), 4526–4530. https://doi.org/10.1021/JACS.8B13042/SUPPL_FILE/JA8B13042_SI_002.PDF"
The structure of flipUM was predicted using alphafold as shown below.
The left shows uncleaved flipUM, the middle the cleaved, active flipUM, and the right regular UM
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 423
Illegal EcoRI site found at 514 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 423
Illegal EcoRI site found at 514 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 423
Illegal EcoRI site found at 514
Illegal BglII site found at 625
Illegal XhoI site found at 682 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 423
Illegal EcoRI site found at 514 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 423
Illegal EcoRI site found at 514
Illegal AgeI site found at 543
Illegal AgeI site found at 789 - 1000COMPATIBLE WITH RFC[1000]
None |