Composite

Part:BBa_K4119008

Designed by: Chengtai Yin   Group: iGEM22_NJTech_China   (2022-09-30)
Revision as of 15:44, 11 October 2022 by L1ndlar (Talk | contribs)


Plac-ΔperR 

   The upper and lower homology arms (HR-UP, HR-DOWN) were constructed at about 500 bp upstream and downstream of the PerR sequence of the Clostridium tyrobutyricum genome, plus a lactose-inducing promoter and a spacer sequence derived from the PerR , generating the CRISPR array .Then inserted it into the pMTL82151 plasmid, the successful plasmid is transformed into E. coli CA434, and introduced into Clostridium tyrobutyricum, by lactose induction to make the spacer hit the perR sequence on the target genome, thereby utilizing the endogenous CRISPR-Cas system knocks the perR out , eliminates the impediment effect of perR on the growth of Clostridium butyrate, thereby achieving aerobic growth of Clostridium tyrobutyricum.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 368
    Illegal PstI site found at 1597
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 368
    Illegal PstI site found at 1597
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 368
    Illegal BglII site found at 428
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 368
    Illegal PstI site found at 1597
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 368
    Illegal PstI site found at 1597
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None