Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Monica Victoria Gutierrez Salazar
Date created: 2016-10-03 11:00:00
Date modified: 2016-10-22 05:13:20
35s + Ga20ox consense + RSIAT-TEV-Luciferase + Tnos
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K2017010_sequence (Version 1) |
Description
Modul of the gRNA testing system created by Valencia UPV 2016 team. This system aims to test the functionality and efficiency of a chosen gRNA to target a particular gene. This system is modular, as the gene inserted can be chosen by the user, only taking into account the required overhangs (5'-AATG-3' and 5'-TTCG-3').It includes a promoter 35s (BBa_K1537015), 20 nucleotides of the Ga20ox consensus gene of Oryza sativa (rice), the reporter luciferase with RSIAT+TEV linker and the terminator Tnos.
Luciferase is out of its reading frame due to a nucleotide added upstream to the linker, and the initial ATG has been removed. This means that when the luciferase is translated, it will not be functional. To make it functional, it is necessary to make indels in the Ga20ox fragment that put the luciferase in the correct reading frame. These indels can be made using the CRISPR/Cas9 system, which is the aim of this device.
Notes
It is important to notice that this device is modular. To insert the region of the gene of interest, this region must have the overhang 5'-AATG-3' and 5'-TTCG-3'.Source
This part was obtained using GoldenBraid assembly. The Ga20ox consense sequence was obtained with oligonucleotide synthesis.| Access | Instance | Definition |
| public public public | BBa_K1537015 BBa_K2017006 BBa_K1484215 | BBa_K1537015 BBa_K2017006 BBa_K1484215 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K1537015 BBa_K2017006 BBa_K1484215 | 1,860 869,2568 2577,2869 | BBa_K1537015 BBa_K2017006 BBa_K1484215 |