Composite

Part:BBa_K737045

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

YtfJ_Comparator

The leading sequence involved in the YtfJ 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 means 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 YtfJ is under control of IPTG inducible device.

2-1.jpg

Gmh2.PNG

The pairing hybridization energy is -12.2 kcal/mol. Although 7 bases are involved in the pairing, microarray analysis indicates that it can give more than 5-fold repression rate and only±0.2 experimental error.

Experimental Data

YTFJ.3.png

Figure 1 ODE simulation of comparator


Ytfj_control.png

Figure 2 Experimental result Figure 3 Expectation effect by ODE simulation

In comparison,we can see YtfJ in accordance with our expectation.


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 1231
    Illegal NheI site found at 1254
    Illegal NheI site found at 2769
    Illegal NheI site found at 2792
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2432
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1181
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3581


[edit]
Categories
Parameters
n/aYtfJ_Comparator