Composite

Part:BBa_K737038

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

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


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,P0412]is a LacI generator, which represses lac promotor and can be induced by IPTG. IPTG 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. 1-3.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 1546
    Illegal NheI site found at 1569
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1208
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2358


Experimental Data

JGE_L1.png

In the picture,L1 represents BBa_ K737038,Jge represents BBa_K737034,DsrA is control.

L1 is composed of J23106, P0412,R0011,Spot42,J23106 and galk::GFP .When there is no IPTG induction,tetR inhibits expression of Spot42 downstream of R0011.Therefore, galk::GFP won't be repressed by Spot42 leading to relatively high expression of GFP.

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