Difference between revisions of "Part:BBa K1351036"

 
 
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<partinfo>BBa_K1351036 short</partinfo>
 
<partinfo>BBa_K1351036 short</partinfo>
  
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Quorum sensing two component system derived from [i]Staphylococcus aureus[/i] N315. This part consists of the AIP-II sensing histidine kinase agrC and the response regulator agrA. Upon detection of autoinducing peptide II (AIP-II), AgrC phosphorylates AgrA, which then bindes to distinct binding sites and activates the P2 and P3 promoters. This part should only work for AIP-II. Other AIPs should lead to inhibition.  
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Quorum sensing two component system derived from ''Staphylococcus aureus'' N315. This part consists of the AIP-II sensing histidine kinase AgrC and the response regulator AgrA. Upon detection of autoinducing peptide II (AIP-II), AgrC phosphorylates AgrA, which then bindes to distinct binding sites and activates the [https://parts.igem.org/Part:BBa_K1351037 P2] and [https://parts.igem.org/Part:BBa_K1351038 P3] promoters. This part should only work for AIP-II. Other AIPs should lead to inhibition.  
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This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix: <br>
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{|
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|prefix with EcoRI, NotI, XbaI, SD and NgoMIV:
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|<span style="color:blue">GAATTC</span><span style="color:green">GCGGCCGC</span>T<span style="color:red">TCTAGA</span>GT<u>AAGGAGG</u>A<span style="color:orange">GCCGGC</span>
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|-
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|suffix with AgeI, SpeI, NotI and PstI:
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|<span style="color:orange">ACCGGT</span><u>TAA</u>T<span style="color:red">AC<u>TAG</u><u>T</u></span><u>A</u><span style="color:green"><u>G</u>CGGCCG</span><span style="color:blue">CTGCAG</span>
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|}
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Sites of restriction enzymes generating compatible overhangs have the same color:<br>
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<span style="color:blue">EcoRI</span> and <span style="color:blue">PstI</span> in blue, <span style="color:green">NotI</span> in green, <span style="color:red">XbaI</span> and <span style="color:red">SpeI</span> in red, <span style="color:orange">NgoMIV</span> and <span style="color:orange">AgeI</span> in orange. Shine-Dalgarno sequence and stop codons are underlined.
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This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus].
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[[File:LMU14 Cap2p3.png|600px]]
  
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Luminescence of ''B. subtilis'' P<sub>''xylA''</sub> - ''agrCA'' / [https://parts.igem.org/Part:BBa_K1351037 P2]-''lux'' and [https://parts.igem.org/Part:BBa_K1351038 P3]-''lux'' was measured in MCSE induced with 0, 0.02, 0.2 % xylose. This plot shows the luminescence readout normalised over OD600 over the course of 6hours. The higher output in xylose induced samples shows the successfull expression of ''agrCA'' in ''B. subtilis''.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 09:21, 27 October 2014

Quorum sensing two component system (AIP-II sensing histidine kinase agrC, response regulator agrA)

Quorum sensing two component system derived from Staphylococcus aureus N315. This part consists of the AIP-II sensing histidine kinase AgrC and the response regulator AgrA. Upon detection of autoinducing peptide II (AIP-II), AgrC phosphorylates AgrA, which then bindes to distinct binding sites and activates the P2 and P3 promoters. This part should only work for AIP-II. Other AIPs should lead to inhibition.

This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:

prefix with EcoRI, NotI, XbaI, SD and NgoMIV: GAATTCGCGGCCGCTTCTAGAGTAAGGAGGAGCCGGC
suffix with AgeI, SpeI, NotI and PstI: ACCGGTTAATACTAGTAGCGGCCGCTGCAG

Sites of restriction enzymes generating compatible overhangs have the same color:
EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.

This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus].

LMU14 Cap2p3.png

Luminescence of B. subtilis PxylA - agrCA / P2-lux and P3-lux was measured in MCSE induced with 0, 0.02, 0.2 % xylose. This plot shows the luminescence readout normalised over OD600 over the course of 6hours. The higher output in xylose induced samples shows the successfull expression of agrCA in B. subtilis.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1139
    Illegal AgeI site found at 1114
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 475