Part:BBa_K206000

Name: pBAD strong Input: [http://openwetware.org/wiki/Arabinose L-arabinose] Output: PoPS

Usage and Biology

pBAD is an E.coli promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC (BBa_I13458) binds to the AraI1 operator site of pBAD and the upstream operator site AraO2, blocking transcription [1]. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription [1].

K206000 is a variant pBAD promoter with a modified AraI1 site that has been shown both to be responsive to lower concentrations of arabinose and to exhibit a higher maximum expression than the wild type (BBa_I13453), as measured by coupling to a fluorescent reporter.

What you can do with it:
BBa_K206000, when used in conjunction with its other family members, can provide varying levels of PoPs output in response to a single level of arabinose input, allowing analog device responses. See [http://2009.igem.org/Team:British_Columbia our wiki] for a project that makes use of this property.

Compatibility:
Chassis: Best used in the E. coli strain [http://cgsc.biology.yale.edu/Strain.php?ID=111773 BW27783], which has been modified to permit homogeneous pBAD promoter expression by substituting the chromosomal arabinose-dependent promoter of AraE (arabinose transporter protein) with a constitutive promoter [2].
Backbone: Has been shown to work on plasmid pSB1C3.
Reporter: Has been shown to work with reporters BBa_I13507 and BBa_I763020.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 125
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 65
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[http://www.ncbi.nlm.nih.gov/pubmed/11102706 [1]] Schlief, R. (2000). Regulation of the L-arabinose operon of Escherichia coli. Trends in Genetics. 16(12):559-565.
[http://www.ncbi.nlm.nih.gov/pubmed/11739756 [2]] Khlebnikov A, Datsenko KA, Skaug T, Wanner BL, and Keasling JD. (2001). Homogeneous expression of the PBAD promoter in Escherichia coli by constitutive expression of the low-affinity high-capacity AraE transporter. Microbiology. 147(12):3241-7.