Plasmid

Part:BBa_K1391138

Designed by: James Anderson   Group: iGEM14_MIT   (2014-10-30)


pEXPR TRE:miRNAg4

To test the effectiveness of this miRNA, we co-transfected pEXPR TRE:miRNA with constitutive pEXPR Hef1a:BACE1. We then induced expression of pEXPR TRE:miRNA by Doxycycline treatment and compared BACE1 expression in the miRNA-transfected HEK293 versus HEK293 that received only Hef1a:BACE1 and no TRE:miRNA. BACE1 activity was quantified using the SensoLyte 520 β-Secretase Assay Kit (Anaspec). BACE1 activity decreased by about 50% when TRE:miRNA was co-transfected.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3087
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3087
    Illegal NheI site found at 2124
    Illegal NheI site found at 2390
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3087
    Illegal BamHI site found at 2554
    Illegal XhoI site found at 2984
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3087
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3087
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3609
    Illegal SapI.rc site found at 2001
    Illegal SapI.rc site found at 2602


[edit]
Categories
Parameters
n/apEXPR TRE:miRNAg4