DNA

Part:BBa_K2351005

Designed by: Ouafa Dahri   Group: iGEM17_Utrecht   (2017-10-23)


Secreted Cas9

note: This part is contains illegal restriction sites.
See BBa_K2351013 for PhytoBrick version and BBa_K2351009 for codon corrected (but unvalidated) BioBrick version.

This sequence codes for the CRISPR-associated protein Cas9 but is being secreted out of the cell due to a signal sequence fused to the N-terminus of the protein. Signal sequence had to be designed to get the protein out of the cell. Also, the sequence has been codon optimized for mammalian cells such that glycosylation has been minimized to preserve protein function. As a result, this biobrick contains illegal restriction-sites. A new version that is compatible has been constructed and has this number: BBa_K2351009. In addition, this brick was uploaded as a phytobrick and the sequence component was validated as BBa_K2351013


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 844
    Illegal PstI site found at 2266
    Illegal PstI site found at 2470
    Illegal PstI site found at 2500
    Illegal PstI site found at 3712
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 844
    Illegal PstI site found at 2266
    Illegal PstI site found at 2470
    Illegal PstI site found at 2500
    Illegal PstI site found at 3712
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 305
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 844
    Illegal PstI site found at 2266
    Illegal PstI site found at 2470
    Illegal PstI site found at 2500
    Illegal PstI site found at 3712
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 844
    Illegal PstI site found at 2266
    Illegal PstI site found at 2470
    Illegal PstI site found at 2500
    Illegal PstI site found at 3712
    Illegal NgoMIV site found at 1132
    Illegal NgoMIV site found at 2236
    Illegal NgoMIV site found at 2309
    Illegal NgoMIV site found at 2794
    Illegal NgoMIV site found at 3703
  • 1000
    COMPATIBLE WITH RFC[1000]


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