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Part:BBa_K737041

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

galK::GFP generator ligated to LacI generator and constitutive sRNA device

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 tet promotor and can be induced by IPTG. aTc concentration gradient results in different sRNA Spot42 levels in cells when presented R0011, which can be monitored by our galK::GFP generator,where galK is the RBS of GFP and the target for Spot42. This device was mistaken for aTc inducible sRNA device originally due to some ligation mistake,but we actually found this one is perfect to express full level sRNA to verify sRNA function without induction.

2-1.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 1545
    Illegal NheI site found at 1568
  • 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 2357


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n/agalK::GFP generator ligated to LacI generator and constitutive sRNA device