Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Cyrille Pauthenier
Date created: 2012-09-17 11:00:00
Date modified: 2015-05-08 01:13:27
GFP-AID OsTirI polysistronic system for auxin detection in tadpole
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K812013_sequence (Version 1) |
Description
This part is the combination of the K81010 (GFP-AID) and K81011 (OsTirI) for co-expression in frogs. It is impossible to create polysistronic mRNA in tadpole. In order to overcome the problem, we fused the proteins with a self-cleavable peptide, Pep2A. The protein is transcribed as a single protein and the the peptide cleave itself or with the help of an endogeneous protease giving the two protein, colocalized in the same cell.This system is a modified version of a pattented device by Kanemaki Masato, Kakimoto Tatsuo, Nishimura Kohei, Takisawa Haruhiko and Fukagawa Tatsuo for yeast and mammalian cells use (http://www.freepatentsonline.com/y2012/0115232.html). However the pattent does not cover the use for oviparian such as frogs and chicken.
This part can be used using micro-injection using for instance a plasmid such as pSC2+ (K82000) or related plasmid.
Notes
This sequence has been assembled using the golden-gate method. Therefore, the biobrick extension are slightely modified, but keep the EcoRI XbaI SpeI PstI system.Source
This device is the reassembly of the parts obtained by PCR in the PNHK60 plasmid from the riken registry.| Sequence Annotation | Location | Component / Role(s) |
| kozac GFP-AID (K81010) Pep2A (self cleavable peptide) OsTirI (K81011) | 1,7 8,1501 1502,1564 1565,3421 | feature/start start_codon feature/protein CDS sequence_feature feature/misc feature/protein CDS |