Difference between revisions of "Part:BBa K1351036"
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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.  
 
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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Revision as of 19:14, 16 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.


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