Part:BBa_K2443037

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 histidine tag. Under the regulation of T7 Promoter, RBS and double terminator.

Original part: <a href="https://parts.igem.org/Part:BBa_I2032" id="pageLink">BBa_I2032</a>

               
Submitted by: MIT
               
Designed by: Bartholomew Canton
               
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 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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Elongation Factor Thermo Stable (EF-Ts)

Original part: <a href="https://parts.igem.org/Part:BBa_K1906000" id="pageLink">BBa_K1906000</a>
Submitted by: XJTLU-China 2016
Designed by: Wenbo Xu
Rational behind improvements: BBa_K1906000 encodes exclusively for the coding region of EF-Ts. 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 C-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Ts 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>). Sequence and Features

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]