Coding

Part:BBa_K1054022

Designed by: Jiasheng Wang   Group: iGEM13_ZJU-China   (2013-09-15)

pLAC promotor,theophylline-inducible Riboswitch(helix-spin), GFP+Terminator

This part is used to standardize the theophylline riboswitch with a helix spin mechanism. It acts differently from another throphylline inducible riboswitch (BBa_K411003).

The protocols of characterization is as follows:

1. Cultivate the BL21 containing BBa_ in LB at 37℃180rpm until OD600 reaches 0.5.

2. Induce the bacteria with 1mM IPTG (final concentration), and cultivate them at 37℃180rpm for 4hrs.

3. Dilute the suspension culture with LB to 0.1OD600 . Theophylline is added at concentration ranging from 1.00E-06 mol/L to1.00E-01 mol/L.

4. Measure the RFU ( ex488/em525) and OD600 synchronously.

5. Data Analysis


500px-ZJU-Standardization 3.1.jpg

Graphic Model about how RFU/OD changes at different time and different concentration of Theophylline.


500px-ZJU-Standardization 3.2.jpg

Graphic Model about how GFP Change Rate changes at different time and different concentration of Theophylline.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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 969


[edit]
Categories
//rna/riboswitch
Parameters
n/apLAC promotor,theophylline-inducible Riboswitch(helix-spin), GFP+Terminator