Coding

Part:BBa_K3902000:Design

Designed by: Florent Poubanne, Enzo Garcia   Group: iGEM21_GO_Paris-Saclay   (2021-10-10)


10His-Tagged-Maltose-binding-protein (MBP)-associated Cas14a1 protein with TEV site


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2590
    Illegal PstI site found at 1380
    Illegal PstI site found at 1546
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2590
    Illegal PstI site found at 1380
    Illegal PstI site found at 1546
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2590
    Illegal BglII site found at 411
    Illegal BamHI site found at 2184
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2590
    Illegal PstI site found at 1380
    Illegal PstI site found at 1546
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2590
    Illegal PstI site found at 1380
    Illegal PstI site found at 1546
    Illegal AgeI site found at 1840
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 109


Design Notes

We needed a sequence for efficient purification of Cas14a1.


Source

This part is from the plasmid pLBH531_MBP-Cas14a1 expression referenced in Addgene as Plasmid #112500 designated by Jennifer Doudna's team and from the publication : Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Harrington LB, Burstein D, Chen JS, Paez-Espino D, Ma E, Witte IP, Cofsky JC, Kyrpides NC, Banfield JF, Doudna JA. Science. 2018 Oct 18. pii: science.aav4294. doi: 10.1126/science.aav4294. 10.1126/science.aav4294 PubMed 30337455

References

Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Harrington LB, Burstein D, Chen JS, Paez-Espino D, Ma E, Witte IP, Cofsky JC, Kyrpides NC, Banfield JF, Doudna JA. Science. 2018 Oct 18. pii: science.aav4294. doi: 10.1126/science.aav4294. 10.1126/science.aav4294 PubMed 30337455