Plasmid

Part:BBa_K1391018

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

pENTR_L7Ae

L7ae is a translational repressor of mRNA transcripts containing the k-turn motif. L7ae/k-turn is a powerful gene repression system.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 239
    Illegal NheI site found at 505
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 116


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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