Part:BBa_K1723007

sgRNA X4 expressing cassette

This part is expressing the sgRNA (single guide RNA) [1] X4 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 X0 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 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_K1723007:Experience

Discover more parts that can work with this one:

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

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 126
  Illegal XhoI site found at 185
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

this part was sent unsequenced

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.