RNA

Part:BBa_K1723003

Designed by: Emilie Cuillery   Group: iGEM15_EPF_Lausanne   (2015-09-15)
Revision as of 17:43, 18 September 2015 by E.Cuillery (Talk | contribs)

sgRNA Z4 expressing cassette

This part is expressing the sgRNA (single guide RNA) [1] Z4 that can form a complex with dCas9-ω (BBa_K1723000). The complex was designed to bind the PAM rich URS J23117 promoter (BBa_K1723001) to activate the production of the target gene linked to the promoter. This part is a part of a gene regulation system built with dCas9-ω, in association with those other parts: sgRNA Z0 expressing cassette (BBa_K1723002) and gRNA Z35 expressing cassette (BBa_K1723004). This sgRNA coding sequence was biobricked with the promoter pBAD and its terminator, both used for our experiments. As the gRNA sequence has to start just after the promoter, the gRNA cannot be put under a promoter using the biobrick assembly system, so we provided a complete expressing cassette but it is still possible to PCR out the part to put it under another promoter. The terminator is specific to sgRNAs and it is recommended to keep it.

Discover more parts that can work with this one:

http://2015.igem.org/Team:EPF_Lausanne/Part_Collection

EPF_Lausanne_Z4.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 185
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 185
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 126
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 185
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 185
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Alec AK Nielsen & Christopher A Voigt (2014). Multi-input CRISPR/Cas circuits that interface host regulatory network. Molecular systems biology, 10(11), 763.


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