Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bob Van Hove, Maarten Van Brempt
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-14 05:25:03
mGFPuv2
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1896001_sequence (Version 1) |
Description
Variant of Green Fluorescent Protein (GFP) that contains the following mutations:- F99S/M153T/V163A: GFPuv or cycle 3 GFP was optimised for excitation by UV light (360-400nm). [1]
- S208L: Almost doubles the fluorescent intensity of GFPuv, resulting in GFPuv2. [2]
- A206K: Monomerizes most GFP variants by disrupting a hydrophobic patch. [3]
Notes
After codon optimisation, 2 BsaI sites were removed.Source
We started from pBAD-GFPuv (GenBank: U62637.1, Clontech), applied the described mutations and codon optimised the sequence for E. coli.References
- von Stetten, D., Noirclerc-Savoye, M., Goedhart, J., Gadella, T. W., & Royant, A. (2012). Structure of a fluorescent protein from Aequorea victoria bearing the obligate-monomer mutation A206K. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 68(8), 878-882.
- Ito, Y., Suzuki, M., & Husimi, Y. (1999). A novel mutant of green fluorescent protein with enhanced sensitivity for microanalysis at 488 nm excitation. Biochemical and biophysical research communications, 264(2), 556-560.
- Crameri, A., Whitehorn, E. A., Tate, E., Stemmer, W. P., Crameri, A., Kitts, P. A., & Kitts, P. A. (1996). Improved green fluorescent protein by molecular evolution using. Nat. Biotechnol, 14(3), 315-319.
| Sequence Annotation | Location | Component / Role(s) |
| start stop mGFPuv2 F99S M153T V163A A206K S208L | 1,3 715,717 1,717 295,297 457,459 487,489 616,618 622,624 | start_codon feature/start feature/stop stop_codon CDS feature/cds feature/mutation sequence_alteration sequence_alteration feature/mutation sequence_alteration feature/mutation feature/mutation sequence_alteration sequence_alteration feature/mutation |