Plasmid

Part:BBa_K5319672:Design

Designed by: YuFei Xie   Group: iGEM24_ZJU-China   (2024-09-30)


lacUV5-J2/3-RS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 425
    Illegal XbaI site found at 843
    Illegal PstI site found at 1209
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1209
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 425
    Illegal XbaI site found at 843
    Illegal PstI site found at 1209
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 425
    Illegal XbaI site found at 843
    Illegal PstI site found at 1209
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2546
    Illegal SapI site found at 1463


Design Notes

The target site of mutation is chosen referring to the idea from "crystal structure of a groun II intron in the pre-catalytic state" to stabilize the RNA in the pre-catalytic state by mutation, which creates a spatial shift in three dimensional structure. The 14-nt recruiting sequence is modified from the sequence in "Hydrolytic endonucleolytic ribozyme (HYER)is programmable for sequence-specific DNA cleavage" corresponding to the site we want it to recognize.


Source

HYER(hydrolytic endonucleolytic ribozymes), identified from group II-C introns in public bacterial genome database in the article "Hydrolytic endonucleolytic ribozyme (HYER)is programmable for sequence-specific DNA cleavage".

References