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Part:BBa_K3776019
INTEGRATE system
Designed by Vo et al. (2020), this system includes a crRNA which guides the TniQ-Cascade and tnsA-tnsB-tnsC operon to insert a donor DNA into a site in an organism's genome. A single promoter drives the expression of the entire system, and can be switched by users. The 32bp crRNA is seamlessly flanked by two repeats and users should use BsaI restriction enzymes to change the crRNA. The TniQ-Cascade complex consists of the proteins TniQ, Cas8, Cas7 and Cas6, which together is guided by the crRNA to search for matching DNA sites and to bind the DNA. After the TniQ-Cascade binds to the DNA and forms an R-loop, TniQ recruits the protein TnsC. Meanwhile, TnsA and TnsB binds and brings together the left and rights ends of the donor DNA and is recruited by TnsC. The donor DNA is flanked by the Right end and Left end, users should clone their genes of interest between these two flanks. The donor DNA is then inserted 49bp
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1004
Illegal BglII site found at 1795
Illegal BglII site found at 2325
Illegal BglII site found at 7570
Illegal BamHI site found at 147
Illegal BamHI site found at 8698
Illegal XhoI site found at 8681 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 7702
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 113
Illegal BsaI.rc site found at 87
Illegal SapI.rc site found at 3081
None |