Composite

Part:BBa_K737044

Designed by: Wenjie Wu and Minghao Gong   Group: iGEM12_OUC-China   (2012-09-19)

NanC_Comparator

The leading sequence involved in the nanC transcript which can act as the target of the sRNA, spot42. It enables strong competition with the galk::GFP to interact with spot42 which shows great potential for constructing sRNA-mediated circuit. This device mean to test feasibility of N/P comparator,replacing Nitrate,Phosphate inputs by IPTG,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.

Spot42 sRNA is under control of aTc inducible device,the buffer RNA NanC is under control of IPTG inducible device.

9-1.PNG

Gmh4.PNG

The pairing hybridization energy is -19.2 kcal/mol. 12 bases are involved in the pairing. Enzymatic assays of nanC-LacZ fusion could give as strong as 47-fold repression. Besides, since nanC’s binding site didn’t coincide with galK’s, Corresponding mutant in spot42 to construct an orthogonal ratio sensor

Experimental Data

Zpr4.jpg

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 1439
    Illegal NheI site found at 1462
    Illegal NheI site found at 2977
    Illegal NheI site found at 3000
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2640
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1261
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1228
    Illegal BsaI.rc site found at 3789


[edit]
Categories
Parameters
n/aNanC_Comparator