Coding

Part:BBa_K1184000:Design

Designed by: Eric Pederson   Group: iGEM13_Carnegie_Mellon   (2013-09-04)

KillerRed

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 151
    Illegal BsaI.rc site found at 442


Design Notes

This sequence is not codon optimized for E. coli. Sequence contains 13 rare proline codons (CCC) and a rare arginine codon (AGA).

KillerRed has catalytic residues E70 and S121. The chromophore forms from the backbone of Q67, Y68 and G69.1,2

KillerRed is derived from the hydrozoan chromoprotein anm2CP. This mutagenesis includes the substitutions T147N and C163A, which are spatially close to the chromophore and drastically affect protein fluorescent properties, as well as the folding- and brightness-improving mutations V23E, D26G, V32I, K75R, N78D, T93S, G116D, I120V, K138R, S156P, Y164F, S177G, L183M, K192E, V201I, I203T, K207R and T222A1,2

For information of the properties of KillerRed, visit our wiki (Carnegie Mellon 2013)

Source

Engineered from anm2CP.1 Provided by the Bruchez lab at Carnegie Mellon.
Genbank: AAY40168

References

1 Bulina, Maria E, Chudakov, Dmitriy M, Britanova, Olga V, Yanushevich, Yurii G, Staroverov, Dmitry B, Chepurnykh, Tatyana V, Merzlyak, Ekaterina M, Shkrob, Maria A, Lukyanov, Sergey, Lukyanov and Konstantin A.
A Genetically Encoded Photosensitizer
Nature Biotechnology 2006 24: 95-99. http://dx.doi.org/10.1038/nbt1175

2 Sergei Pletnev, Nadya G. Gurskaya, Nadya V. Pletneva, Konstantin A. Lukyanov, Dmitri M. Chudakov, Vladimir I. Martynov, Vladimir O. Popov, Mikhail V. Kovalchuk, Alexander Wlodawer, Zbigniew Dauter, and Vladimir Pletnev.
Structural Basis for Phototoxicity of the Genetically Encoded Photosensitizer KillerRed
Journal of Biological Chemistry 2009 284: 32028-32039. First Published on September 8, 2009, doi:10.1074/jbc.M109.054973

3Molecular graphics and analyses were performed with the UCSF Chimera package. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, funded by grants from the National Institutes of Health National Center for Research Resources (2P41RR001081) and National Institute of General Medical Sciences (9P41GM103311).

4Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE.
UCSF Chimera--a visualization system for exploratory research and analysis.
Journal of Computational Chemistry. 2004 Oct;25(13):1605-12.