Plasmid
Part:BBa_K5319672:Design
Designed by: YuFei Xie Group: iGEM24_ZJU-China (2024-09-30)
lacUV5-J2/3-RS
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 425
Illegal XbaI site found at 843
Illegal PstI site found at 1209 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1209
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 425
Illegal XbaI site found at 843
Illegal PstI site found at 1209 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 425
Illegal XbaI site found at 843
Illegal PstI site found at 1209 - 1000INCOMPATIBLE 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".