Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Matthew Polasko
Date created: 2010-09-27 11:00:00
Date modified: 2015-05-08 01:12:26
Strong Plant RBS (Kozak sequence) + mCherry from J06504
| Types | DnaRegion |
| Roles | engineered_region Reporter |
| Sequences | BBa_K414003_sequence (Version 1) |
Description
In order for mCherry Fluorescent Protein to function properly in plants, a plant-specific ribosome binding site (Kozak sequence)must be added to the 5'region just upstream from the start of translation.The Kozak sequence is AAA AAA AAA ACA and is directly adjacent to the ATG start codon in mCherry.One point mutation that should be noted is nucleotide 483 of J06504 (mCherry). C -> T is the mutation, but examining the translation of the sequence in the proper reading frame yields no translational discrepancies. Also, no illegal iGEM restriction sites were generated from the point mutation.
Notes
One consideration we made was the strength of the RBS we wanted upstream of the reporter. Evidence can be found in the literature that a spectrum of translational activity can be generated by tampering with the Kozak sequence in plants. Future teams may wish to investigate how they can tamper with their Kozak of designing a system that requires discrete but relative expression of genes initiated by the same promoters.Source
The source of mCherry was derived using modified primers containing the Kozak sequence designed to bind to the iGEM part BBa_J06702 (Wang,2005)| Sequence Annotation | Location | Component / Role(s) |
| Kozak Kpn I Translational Start mCherry (J06504) Translational Stop 2x C -> T | 7,18 1,6 19,19 19,726 727,727 501,501 | ribosome_entry_site feature/rbs sequence_feature feature/misc feature/start start_codon feature/cds CDS stop_codon feature/stop feature/mutation sequence_alteration |