Part:BBa_K845000
iGEM 2012 Grenoble Team proposes a new design and application to the pBAD promoter(paraBAD).
In order to create a biological AND gate the 2012 Grenoble team worked on the cAMP-CRP complex and arabinose inductible promoter.
The activity of the promoter was tested under several conditions ( http://2012.igem.org/Team:Grenoble/Biology/Protocols/AND_test ) using a GFP as a reporter (on a pUA66). We tested this activity in BW25113 ΔcyaA in different culture media (Glucose + Acetate; Glycerol; Acetate).
Usage and Biology
paraBAD is a promoter which can be activated by 2 molecules : CRP-cAMP complex and the AraC protein. It has two states; in absence of L-arabinose the paraBAD promoter is repressed by AraC, whereas the paraC promoter is activated (unless an excess of AraC is present). In presence of L-arabinose and the CRP-cAMP complex, the promoter is activated thus enabling the transcription of the downstream elements. This AND gate provides a filter to biological noise.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 500
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 344
Illegal XhoI site found at 494 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 500
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 500
Illegal AgeI site found at 179
Illegal AgeI site found at 484 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1169
Illegal SapI site found at 161
//promoter
| emission | 504 nm |
| excitation | 395 and 475 nm |
| negative_regulators | AraC protein |
| positive_regulators | cAMP-CRP complex and Arabinose |