Difference between revisions of "Part:BBa K3776019"
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<partinfo>BBa_K3776019 short</partinfo>
 
<partinfo>BBa_K3776019 short</partinfo>
  
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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. This RNA guided protein complex inserts a donor DNA located between two flanks behind the into the genome of the host organism. Users should clone their genes of interest between these two flanks.  
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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
  
 
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Revision as of 02:55, 18 October 2021


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1578
    Illegal NheI site found at 3665
    Illegal NheI site found at 7783
  • 21
    INCOMPATIBLE 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
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 7702
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 113
    Illegal BsaI.rc site found at 87
    Illegal SapI.rc site found at 3081