Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Monica Victoria Gutierrez Salazar
Date created: 2016-10-02 11:00:00
Date modified: 2016-10-22 07:17:21
RSIAT-TEV linker + Luciferase
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K2017006_sequence (Version 1) |
Description
Firefly luciferase protein. It oxidates its substrate luciferin, emitting light during the process. It can act as a reporter protein when used with a promoter. This part is made to be fused downstream to other coding sequence, so it includes the RSIAT-TEV linker in order to let the luciferase acquire the correct tertiary structure. The TEV region is the recognition site for the NiaTEV protease. The protease cuts on its site, letting the luciferase without the upstream coding sequence.The luciferase does not include ATG (Met) to initiate translation, as it needs to be fused to other coding sequence on the 5'. The translation must begin in the coding sequence fused upstream. The linker includes a random nucleotide in 5', to change the reading frame of the luciferase. That way, it will not be translated unless an indel is produced in the coding region to which the part is fused.
Notes
This part must be fused downstream to other coding sequence. In order to be used in our modular gRNA testing system, it was necessary to remove the ATG (Met codon) to avoid unexpected translation. The translation must begin in the coding sequence fused upstream. The linker includes a random nucleotide in 5', to change the reading frame of the luciferase. That way, it will not be translated unless an indel is produced in the coding region to which the part is fused.Source
Luciferase was obtained from laboratory stock. The linker was fused with a PCR.| Sequence Annotation | Location | Component / Role(s) |
| Additional nucleotide RSIAT Linker Luciferase | 1,2 3,19 20,1700 | feature/misc sequence_feature feature/misc sequence_feature CDS feature/protein |