Difference between revisions of "Part:BBa K1723006"
 
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<partinfo>BBa_K1723006 short</partinfo>
 
<partinfo>BBa_K1723006 short</partinfo>
  
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This part is expressing the sgRNA (single guide RNA) X0 that can bind to dCas9-ω (BBa_K1723000). The complex was designed to bind the PAM rich URS J23117Alt promoter (BBa_K1723005) to repress the production of the target gene linked to the promoter. This part is a part of a gene regulation system built with dCas9-ω, with the parts sgRNA X4 expressing cassette (BBa_K1723006) and gRNA X35 expressing cassette (BBa_K1723008). 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 the first base after the promoter it couldn't have been used with the standard biobrick system so the promoter is provided already. It is possible to PCR out the part to put it under another promoter, but he terminator is specific and it is recommended to keep it.
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This part is expressing the sgRNA (single guide RNA) [1] X0 that can form a complex with dCas9-ω (BBa_K1723000). The complex was designed to bind the PAM rich URS J23117Alt promoter (BBa_K1723005) to repress 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 X4 expressing cassette (BBa_K1723007) and gRNA X35 expressing cassette (BBa_K1723008). 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.
  
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Discover all the parts that can work with this one:
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<b>This part was experimentally validated, see </b>https://parts.igem.org/Part:BBa_K1723006:Experience
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Discover more parts that can work with this one:
  
 
http://2015.igem.org/Team:EPF_Lausanne/Part_Collection
 
http://2015.igem.org/Team:EPF_Lausanne/Part_Collection
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<partinfo>BBa_K1723002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1723002 SequenceAndFeatures</partinfo>
  
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===References===
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[1] Alec AK Nielsen & Christopher A Voigt (2014). Multi-input CRISPR/Cas circuits that interface host regulatory network. Molecular systems biology, 10(11), 763.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 00:50, 19 September 2015

sgRNA X0 expressing cassette

This part is expressing the sgRNA (single guide RNA) [1] X0 that can form a complex with dCas9-ω (BBa_K1723000). The complex was designed to bind the PAM rich URS J23117Alt promoter (BBa_K1723005) to repress 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 X4 expressing cassette (BBa_K1723007) and gRNA X35 expressing cassette (BBa_K1723008). 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.

This part was experimentally validated, see https://parts.igem.org/Part:BBa_K1723006:Experience

Discover more parts that can work with this one:

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

EPF_Lausanne_Z0.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 179
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 126
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 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.