Composite

Part:BBa_K3081061

Designed by: Xiaoying Qiao   Group: iGEM19_Peking   (2019-10-16)


dCas9 J23119 PolyA with the thermo-sensitive transcription factor TCI42

This part contains the coding gene of the thermo-sensitive transcription factor TcI42 under the constitutive promoter J23119. TcI42 is an upshifted variant of a well-known temperature-sensitive variant of the bacteriophage λ repressor cI (mutant cI857, here referred to as TcI) acting on a tandem pR–pL operator–promoter. The promoter induces the expression of CRISPRri system(BBa_K3081007), including dCas9 and sgRNA. The sgRNA targets a 20bp DNA sequence in E.coli OriC, which will arrest and inhibit the genome replication initiation.But in this composite part, the sgRNA polyA is used as control. The right target sgRNA “M+” is described at BBa_K3081059.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 753
    Illegal NheI site found at 776
    Illegal NheI site found at 5286
    Illegal NheI site found at 5309
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1297
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None