Difference between revisions of "Part:BBa K1723003"
 
(3 intermediate revisions by the same user not shown)
Line 2: Line 2:
 
<partinfo>BBa_K1723003 short</partinfo>
 
<partinfo>BBa_K1723003 short</partinfo>
  
−
This part is expressing the sgRNA (single guide RNA) Z4 that can bind to dCas9-&#969; (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-&#969;, with the 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 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.
+
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 all the parts that can work with this one:
+
<b>This part was experimentally validated, see </b>https://parts.igem.org/Part:BBa_K1723003:Experience
 +
 
 +
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
Line 17: Line 19:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1723003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1723003 SequenceAndFeatures</partinfo>
 +
 +
===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.
  
  

Latest revision as of 00:51, 19 September 2015

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.

This part was experimentally validated, see https://parts.igem.org/Part:BBa_K1723003:Experience

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.