Team:EPF Lausanne/Part Collection
E. coli.
dCas9-ω
dCa9s-ω is our main tool to build our project in E. coli. This catalytically dead Cas9 fused to the RNA polymerase ω subunit can act as an activator or as an inhibitor when provided the right guide RNA.
PAM rich URS J23117
This promoter is composed of the weak J23117 promoter, BBa_J23117, fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind as dCas9 cannot bind without a PAM sequence.
sgRNA Z0
the sgRNA (single guide-RNA) Z0 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA Z4
the sgRNA Z4 activates PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA Z35
the sgRNA Z35 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
PAM rich URS J23117Alt
This promoter was made on the same template as PAM rich URS J23117 promoter, but was mutated to obtain different sequences for sgRNA targeting. It is also fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind, as dCas9 cannot bind without a PAM sequence.
sgRNA X0
the sgRNA (single guide-RNA) X0 inhibit PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA X4
the sgRNA X4 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA X35
the sgRNA X35 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
