Difference between revisions of "Part:BBa K1897008"
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<partinfo>BBa_K1897008 short</partinfo>
 
<partinfo>BBa_K1897008 short</partinfo>
  
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This part was made up of promoter <partinfo>BBa_J23119</partinfo>, RBS <partinfo>BBa_B0030</partinfo>, LuxR coding sequence <partinfo>BBa_C0062</partinfo> and double terminator <partinfo>BBa_B0015</partinfo>.
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This part was made up of promoter <partinfo>BBa_J23119</partinfo>, ribosome binding site (RBS) <partinfo>BBa_B0030</partinfo>, LuxR coding sequence <partinfo>BBa_C0062</partinfo> and double terminator <partinfo>BBa_B0015</partinfo>.
  
 
===Usage and Biology===
 
===Usage and Biology===
 
This part allows for the constitutive expression of LuxR protein from the promoter <partinfo>BBa_J23119</partinfo>. The LuxR protein originated from <i>Vibrio fischeri</i>, and is a regulatory protein of the Lux operon. The Lux operon can be transcribed from from te left (operonL) or right (operonR). Transcription of operonR, which consists of LuxICDABE, occurs only in the presence of N-(3-oxo-hexanoyl)-homoserine lactone (AHL), which is an autoinducer produced by LuxI (Shaldel and Baldwin, 1991). This also produces light due to the production of LuxCDABE (Shaldel and Baldwin, 1991). Transcription of operonL leads to the production of LuxR (Shaldel and Baldwin, 1991). In the presence of rising amounts of AHL, LuxR-AHL complexes begin to form, and LuxR undergoes a conformational change that leads to the activation of the Lux operon (Trott, 2000).  
 
This part allows for the constitutive expression of LuxR protein from the promoter <partinfo>BBa_J23119</partinfo>. The LuxR protein originated from <i>Vibrio fischeri</i>, and is a regulatory protein of the Lux operon. The Lux operon can be transcribed from from te left (operonL) or right (operonR). Transcription of operonR, which consists of LuxICDABE, occurs only in the presence of N-(3-oxo-hexanoyl)-homoserine lactone (AHL), which is an autoinducer produced by LuxI (Shaldel and Baldwin, 1991). This also produces light due to the production of LuxCDABE (Shaldel and Baldwin, 1991). Transcription of operonL leads to the production of LuxR (Shaldel and Baldwin, 1991). In the presence of rising amounts of AHL, LuxR-AHL complexes begin to form, and LuxR undergoes a conformational change that leads to the activation of the Lux operon (Trott, 2000).  
  
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This part can be used if you need constitutive production of LuxR to activate production of genes under the control of pLuxR (<partinfo>BBa_R0062</partinfo>).
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This part can be used if you require constitutive production of LuxR to activate production of genes under the control of pLuxR (<partinfo>BBa_R0062</partinfo>).
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1897008 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1897008 SequenceAndFeatures</partinfo>
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===Construction of composite LuxR===
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The full composite LuxR was made from overlap PCR of 3 fragments - namely the "front" fragment, comprising of the biobrick prefix, promoter, RBS, as well as some bases from the LuxR coding sequence; the "middle" fragment, comprising of a little of the RBS, the coding sequence of LuxR, the HA tag as well as a few bases of the terminator sequence; and the "terminator" fragment, comprising of a small part of the LuxR coding sequence, the HA tag, the double terminator and the biobricks suffix.
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Revision as of 16:08, 10 October 2016


LuxR

This part was made up of promoter BBa_J23119, ribosome binding site (RBS) BBa_B0030, LuxR coding sequence BBa_C0062 and double terminator BBa_B0015.

Usage and Biology

This part allows for the constitutive expression of LuxR protein from the promoter BBa_J23119. The LuxR protein originated from Vibrio fischeri, and is a regulatory protein of the Lux operon. The Lux operon can be transcribed from from te left (operonL) or right (operonR). Transcription of operonR, which consists of LuxICDABE, occurs only in the presence of N-(3-oxo-hexanoyl)-homoserine lactone (AHL), which is an autoinducer produced by LuxI (Shaldel and Baldwin, 1991). This also produces light due to the production of LuxCDABE (Shaldel and Baldwin, 1991). Transcription of operonL leads to the production of LuxR (Shaldel and Baldwin, 1991). In the presence of rising amounts of AHL, LuxR-AHL complexes begin to form, and LuxR undergoes a conformational change that leads to the activation of the Lux operon (Trott, 2000).

This part can be used if you require constitutive production of LuxR to activate production of genes under the control of pLuxR (BBa_R0062).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 50
  • 1000
    COMPATIBLE WITH RFC[1000]

Construction of composite LuxR

The full composite LuxR was made from overlap PCR of 3 fragments - namely the "front" fragment, comprising of the biobrick prefix, promoter, RBS, as well as some bases from the LuxR coding sequence; the "middle" fragment, comprising of a little of the RBS, the coding sequence of LuxR, the HA tag as well as a few bases of the terminator sequence; and the "terminator" fragment, comprising of a small part of the LuxR coding sequence, the HA tag, the double terminator and the biobricks suffix.