Part:BBa_J10069
EPIC Firefly Luciferase and LRE P. pyralis + BD18
EPIC Firefly Luciferase and LRE from P. Pyralis (E. coli optimised)
This is a modification of part K325100 originally built by iGEM10_Cambridge. We have removed the initial RBS to facilitate translation using C-dog bicistronic linker BD18 (J119024).
This part is a translational unit for a mutant of the luciferase from the North American firefly (P. pyralis) as well as this species' luciferin regenerating enzyme (LRE). D-Luciferin has to be added to obtain light output. EPIC stands for Enhanced Photon Initiating Complex. It is described in Fujii et al. 2007 as having a 10 times higher substrate affinity and luminescence output compared to wildtype. The part is codon optimised for expression in E.coli.
References [1]: S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,Life 61, 6-17. [2]: T. Nakatsu et al. (2006) Structural Basis for the spectral difference in luciferase bioluminescence, Nature 440(16), 372-376. [3]: K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, The Journal of Biological Chemistry, 276(39), 36508-36513.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal suffix found in sequence at 2624
Illegal EcoRI site found at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 2625
Illegal PstI site found at 2639
Illegal NotI site found at 2632 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
Illegal BglII site found at 2577
Illegal BamHI site found at 945 - 23INCOMPATIBLE WITH RFC[23]Illegal suffix found in sequence at 2625
Illegal EcoRI site found at 1 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1
Illegal SpeI site found at 2625
Illegal PstI site found at 2639 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 7
Illegal SapI.rc site found at 2281
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