Coding

Part:BBa_K3001004

Designed by: Sydnee Calhoun   Group: iGEM19_Lethbridge_HS   (2019-10-14)


Cas13a Lwa Coding Sequence + purification tags

This protein is an RNA cleaving enzyme called Cas13a that was isolated from Leptotrichia wadei (Lwa). It interacts with a direct repeat stem loop on a CRISPR RNA (crRNA) that contains a specific sequence for a target RNA transcript. This construct is designed to be able to be expressed under an inducible promoter (T7). The protein itself has several purification tags (His, maltose binding protein and a TEV cleavage site to remove the purification tags). This construct was ordered from Addgene. Lwa was previously submitted by Munich 2017 (BBa_K2323001). We have improved this part by having this construct in a fully inducibly expressible form (T7 promoter and ribosomal binding site present) and varying the purification tags that future iGEM teams can use when working with this protein.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 163
    Illegal EcoRI site found at 1636
    Illegal EcoRI site found at 1761
    Illegal EcoRI site found at 1864
    Illegal EcoRI site found at 2155
    Illegal EcoRI site found at 2371
    Illegal EcoRI site found at 2413
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 163
    Illegal EcoRI site found at 1636
    Illegal EcoRI site found at 1761
    Illegal EcoRI site found at 1864
    Illegal EcoRI site found at 2155
    Illegal EcoRI site found at 2371
    Illegal EcoRI site found at 2413
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 163
    Illegal EcoRI site found at 1636
    Illegal EcoRI site found at 1761
    Illegal EcoRI site found at 1864
    Illegal EcoRI site found at 2155
    Illegal EcoRI site found at 2371
    Illegal EcoRI site found at 2413
    Illegal BglII site found at 299
    Illegal BglII site found at 581
    Illegal BglII site found at 842
    Illegal BglII site found at 1370
    Illegal BglII site found at 2090
    Illegal BglII site found at 3401
    Illegal BglII site found at 3497
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 163
    Illegal EcoRI site found at 1636
    Illegal EcoRI site found at 1761
    Illegal EcoRI site found at 1864
    Illegal EcoRI site found at 2155
    Illegal EcoRI site found at 2371
    Illegal EcoRI site found at 2413
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 163
    Illegal EcoRI site found at 1636
    Illegal EcoRI site found at 1761
    Illegal EcoRI site found at 1864
    Illegal EcoRI site found at 2155
    Illegal EcoRI site found at 2371
    Illegal EcoRI site found at 2413
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None