Coding

Part:BBa_K2316001

Designed by: Shaw Kar Ming   Group: iGEM17_NTU_SINGAPORE   (2017-10-23)
Revision as of 11:32, 30 October 2017 by Dannyteo92 (Talk | contribs) (→‎Usage and Biology)


∆RuvCIII-2 ∆HNH ∆REC2 Sp-dCas9 Enhanced

dCas9 is a catalytically inactive Cas9, which still retains it's ability to bind to DNA. For epigenetic regulation, the dCas9 is highly dependent upon the function of it's fusion partner. This is an improvement from part BBa_K2136000.


Usage and Biology

For this part, we have deleted the HNH domain, Rec2 domain and the RuvCIII-2 region of the SP-dCas9. Quick changes are further done at Asn497Lys, Thr657Lys and Ser1109Arg. It's binding capability was tested by assessing the strength in transcriptional activation via by tagging it with a VP64-p65-Rta tripartite activator. The three quick changes show improvement in the binding capability SP-dCas9.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 251
    Illegal BglII site found at 804
    Illegal BamHI site found at 1622
    Illegal XhoI site found at 2578
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1966
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2112
    Illegal BsaI site found at 2774
    Illegal BsaI.rc site found at 1027


[edit]
Categories
Parameters
None