Device

Part:BBa_K2017013

Designed by: Monica Victoria Gutierrez Salazar   Group: iGEM16_Valencia_UPV   (2016-10-04)
Revision as of 15:08, 4 October 2016 by Registry (Talk | contribs)

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35s + TFL consense + AEK-Luciferase + Tnos

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 TFL consensus gene of Clemenules (clementine orange), the reporter luciferase with AEK 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 TFL 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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 66
    Illegal BsaI site found at 2580
    Illegal BsaI.rc site found at 2867
    Illegal SapI.rc site found at 1726


[edit]
Categories
//cds/reporter
//chassis/eukaryote/nbenthamiana
//function/crispr
Parameters
None