Plasmid

Part:BBa_K1391019

Designed by: Jing Wei "Raymond" Liu   Group: iGEM14_MIT   (2014-10-17)

MAV1212_hEF1a_L7Ae

L7ae is constitutively expressed under a hEF1a promoter, within a MAV backbone. Simple complete module for L7ae expression.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal EcoRI site found at 3025
    Illegal XbaI site found at 2998
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal PstI site found at 2986
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal EcoRI site found at 3025
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal PstI site found at 2986
    Illegal NotI site found at 402
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal EcoRI site found at 3025
    Illegal BglII site found at 1694
    Illegal BglII site found at 3486
    Illegal BamHI site found at 3004
    Illegal XhoI site found at 394
    Illegal XhoI site found at 2093
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal EcoRI site found at 3025
    Illegal XbaI site found at 2998
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal PstI site found at 2986
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal EcoRI site found at 3025
    Illegal XbaI site found at 2998
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal PstI site found at 2986
    Illegal NgoMIV site found at 645
    Illegal NgoMIV site found at 926
    Illegal NgoMIV site found at 1828
    Illegal AgeI site found at 774
    Illegal AgeI site found at 1206
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2769
    Illegal BsaI.rc site found at 3387


Before coming to any conclusions about the success of our constructs, we needed to make sure that the L7ae/k-turn system worked correctly. To do this we expressed k-turn:eGFP with and without the presence of constitutive L7ae. We used eBFP as our normalizing transfection marker.


L7ae Repression of 2x-Kturn-GFP. In the absence of L7ae, eBFP2 and GFP fluorescence show a positive relationship, as expected when co-expressing two constitutive fluorophores. However, upon the addition of L7ae into the system, GFP production is almost completely silenced. This indicates that our L7ae / 2x-Kturn system is viable to implement in our high sensors.

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Categories
Parameters
None