Composite

Part:BBa_K737039

Designed by: Wenjie Wu and Peiran Zhang   Group: iGEM12_OUC-China   (2012-09-19)

galK::GFP generator ligated to sRNA device controlled by aTc

Small RNA Spot42 controls the GFP mRNA translation by blocking the RBS in galK.The galK and E0040 is fused together using ClaI site to avoid stop condon in standard scar.J23106 is a moderate promotor,and galK’s RBS is a moderate one.

[J23106,P0440]is a TetR generator, which represses tet promotor and can be induced by aTc. aTc concentration gradient results in different sRNA Spot42 levels in cells, which can be monitored by our galK::GFP generator,where galK is the RBS of GFP and the target for Spot42.

9-1.PNG


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1077
    Illegal NheI site found at 1100
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1889


Experimental Data

JGE_L2.png

In the picture,L2 represents BBa_ K737039,JgE represents BBa_K737034,DsrA is control.

L2 consists of J23106, P0440,R0040,Spot42,J23106 and galk::GFP .When there is no aTc induction,LacI inhibits expression of Spot42 downstream of R0040.Therefore, galk::GFP won't be repressed by Spot42 leading to relatively high expression of GFP.


K737039.jpg

In the picture we can see the satisfying inductive effect of aTc.

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n/agalK::GFP generator ligated to sRNA device controlled by aTc