Difference between revisions of "Part:BBa K2443037"
 
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<partinfo>BBa_K2443037 short</partinfo>
 
<partinfo>BBa_K2443037 short</partinfo>
  
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T7 RNA Polymerase is responsible for transcribing DNA into RNA and is under the regulation of the T7 promoter consensus sequence. It has been Codon optimized for use in <i>Escherichia coli</i> and contains a N-terminal histidine tag. Under the regulation of T7 Promoter, RBS and double terminator.  
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T7 RNA Polymerase is responsible for transcribing DNA into RNA and is under the regulation of the T7 promoter consensus sequence. It has been Codon optimized for use in <i>Escherichia coli</i> and contains a N-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator.  
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<h1> Improved Part </h1>
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<p><b>Original part:</b>BBa_I2032 submitted by MIT 2006</p>
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<p><b> Rational behind improvements:</b>  BBa_I2032 encodes exclusively for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our Next vivo system we have Codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized our T7 RNA polymerase to be overexpressed in BL21-Gold (DE3) cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015). </p>
  
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<h1> Improved part: </h1>
 
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<p class="text12"> <b> Original part:</b> <a href="https://parts.igem.org/Part:BBa_I2032" id="pageLink">BBa_I2032</a>
 
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                <br><b>Submitted by:</b> MIT
 
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                <br><b>Designed by:</b> Bartholomew Canton
 
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                <br><b>Rational behind improvements:</b> BBa_I2032 encodes exclusively for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our N<i>ex</i>t <i>vivo</i> system we have Codon optimized for use in <i>E. coli</i> and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized our T7 RNA polymerase to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (<a href="https://parts.igem.org/Part:BBa_I719005" id="pageLink">BBa_I719005</a>), RBS (<a href="https://parts.igem.org/Part:BBa_B0034" id="pageLink">BBa_B0034</a>) and double terminator (<a href="https://parts.igem.org/Part:BBa_B0015" id="pageLink">BBa_B0015</a>).
 
  
 
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===Usage and Biology===
 
===Usage and Biology===
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<p>
 
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>Sequence and Features</span></p>
 
<partinfo>BBa_K2443037 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2443037 SequenceAndFeatures</partinfo>
  

Latest revision as of 00:12, 1 November 2017


T7 RNA Polymerase optimized for expression in E. coli

T7 RNA Polymerase is responsible for transcribing DNA into RNA and is under the regulation of the T7 promoter consensus sequence. It has been Codon optimized for use in Escherichia coli and contains a N-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator.

Improved Part

Original part:BBa_I2032 submitted by MIT 2006

Rational behind improvements: BBa_I2032 encodes exclusively for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our Next vivo system we have Codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized our T7 RNA polymerase to be overexpressed in BL21-Gold (DE3) cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).


Sequence and Features</p>


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1719
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 275
    Illegal NgoMIV site found at 1496
    Illegal NgoMIV site found at 1943
  • 1000
    COMPATIBLE WITH RFC[1000]