BBa_K2017010

BBa_K2017010 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2017010
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.

igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K2017010/1