Composite

Part:BBa_K3081016

Designed by: Yiwen Wang   Group: iGEM19_Peking   (2019-10-16)


pBAD-dCas9-J23119-M-R2+

This composite part is the principal design of the inducible CRISPR-based DNA replication interference system, with the 20 bp sgRNA targeting to the linker sequence of two DnaA binding boxes (M and R2) E.coli genome replication initiation region, OriC. Not considered to be DnaA binding box,blocking this site has no inhibition on cell growth.

Experiment

For more detailed information, see BBa_K3081058


Reference:

[1]Wolański M, Donczew R, Zawilakpawlik A, et al. oriC-encoded instructions for the initiation of bacterial chromosome replication.[J]. Frontiers in Microbiology, 2015, 5(735):735.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal NheI site found at 5491
    Illegal NheI site found at 5514
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1486
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


[edit]
Categories
Parameters
None