Difference between revisions of "Part:BBa K2066030"
 
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This part was designed to be assembled via Gibson Assembly from WM16_014 (using primers WM16_P027 and WM16_P009) and the same WM16_014 again (using primers WM16_P019 and WM16_P025).
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__NOTOC__
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<partinfo>BBa_K2066030 short</partinfo>
  
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This part follows Lou et al. 2012 (“Ribozyme-based insulator parts buffer synthetic circuits from genetic context”). This part will be used in concert with WM16_031 (pTAC Ribozyme Chracterization Part – cI GFP; no RiboJ) to determine the influence of the RiboJ insulator sequence on gene expression.
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This part is used for the 2016 William and Mary iGEM Ribozyme Characterization project.  
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Choice of promoter influences the composition of the 5’ UTR on an RNA transcript. These 5’ UTR sequences can influence translation efficiency in a coding sequence-dependent manner. In 2012 Lou and Colleagues from the Voigt lab at MIT proposed a solution. By introducing the self-cleaving RNA ribozyme RiboJ immediately upstream of the RBS the  authors were able to standardize the 5’ UTR of their transcripts.
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We wanted to adapt this system to our Circuit Control Toolbox in order to insulate the input-output relationship (transfer function) of a circuit from its coding sequence. This allows our circuit modifications to be made independent of the original circuit architecture.  
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This part is a member of a set of test characterization devices to ensure that RiboJ insulation in the BioBrick standard insulates genetic circuitry. The part can be used in combination with BBa_K2066016 (constitutive Lac repressor) to investigate the transfer function.
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For more information please visit our wiki: http://2016.igem.org/Team:William_and_Mary/RiboJ
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References:
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Lou, Chunbo, et al. "Ribozyme-based insulator parts buffer synthetic circuits from genetic context." Nature biotechnology 30.11 (2012): 1137-1142.
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<!-- -->
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2066030 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K2066030 parameters</partinfo>
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<!-- -->

Latest revision as of 05:17, 29 October 2016

pTAC sfGFP

This part is used for the 2016 William and Mary iGEM Ribozyme Characterization project.

Choice of promoter influences the composition of the 5’ UTR on an RNA transcript. These 5’ UTR sequences can influence translation efficiency in a coding sequence-dependent manner. In 2012 Lou and Colleagues from the Voigt lab at MIT proposed a solution. By introducing the self-cleaving RNA ribozyme RiboJ immediately upstream of the RBS the authors were able to standardize the 5’ UTR of their transcripts.

We wanted to adapt this system to our Circuit Control Toolbox in order to insulate the input-output relationship (transfer function) of a circuit from its coding sequence. This allows our circuit modifications to be made independent of the original circuit architecture.

This part is a member of a set of test characterization devices to ensure that RiboJ insulation in the BioBrick standard insulates genetic circuitry. The part can be used in combination with BBa_K2066016 (constitutive Lac repressor) to investigate the transfer function.

For more information please visit our wiki: http://2016.igem.org/Team:William_and_Mary/RiboJ

References: Lou, Chunbo, et al. "Ribozyme-based insulator parts buffer synthetic circuits from genetic context." Nature biotechnology 30.11 (2012): 1137-1142.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 132