Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Malcolm Campbell
Date created: 2012-01-10 12:00:00
Date modified: 2015-08-31 04:08:24
EPIC Firefly Luciferase and LRE P. pyralis + BD18
| Types | DnaRegion |
| Roles | engineered_region Reporter |
| Sequences | BBa_J10069_sequence (Version 1) |
Description
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.
Notes
removed RBS from J119024Source
from registry parts, synthesized de novo| Sequence Annotation | Location | Component / Role(s) |
| GGA prefix AATG sticky word BD18 ATG luciferin regenerating enzyme double stop ATG EPIC luciferase double stop BBa suffix | 1,17 15,17 15,944 939,944 968,970 968,2623 2618,2623 2624,2644 | sequence_feature feature/dna feature/start start_codon CDS feature/protein feature/stop stop_codon start_codon feature/start feature/protein CDS feature/stop stop_codon sequence_feature feature/misc |