Difference between revisions of "Part:BBa K3122001"
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<partinfo>BBa_K3122001 short</partinfo>
 
<partinfo>BBa_K3122001 short</partinfo>
  
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This part is the constitutive medium promoter, from Anderson family, was adapted to type IIS enzymes standard.
  
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This part is derived from BBa_K1223006. It contains 6 histidine residues, and it has been adapted to AEGIS' lamB display system in order to be used as the heterologous peptide tag expressed.  
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It has been used to express our Level 1 constructs related to membrane proteins. The reason behind this choice relays on its relative strength among other members of the family: it is a medium- strength regulator (0.36) which should not express too much membrane protein as to induce cell lysis.  
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To see the design of this part go to the Design tab.
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<h2> Characterization </h2>
 
<h2> Characterization </h2>
  
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Characterization of this part has been conducted by building a transcriptional unit <partinfo>BBa_K3122004</partinfo> and performing assays with nickel beads in order to check the interaction between them and its force. This transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:
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Characterization of this part has been conducted by building two transcriptional units (BBa_K3122004, BBa_ K3122005), and performing Lambda fague and Cell separation assays.
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BBa_K3122004 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:
 
<ul>
 
<ul>
 
<li><partinfo>BBa_K3122003</partinfo>: Promoter J23107 adapted to type IIS assembly</li>
 
<li><partinfo>BBa_K3122003</partinfo>: Promoter J23107 adapted to type IIS assembly</li>
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</ul>
 
</ul>
  
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<h2> How to use it </h2>  
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BBa_K3122005 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:
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<ul>
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<li><partinfo>BBa_K3122003</partinfo>: Promoter J23107 adapted to type IIS assembly</li>
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<li><partinfo>BBa_K2656009</partinfo>: RBS  B0030 adapted to type IIS assembly</li>
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<li><partinfo>BBa_K3122000</partinfo>: LamB coding sequence. This part is our AEGIS' lamB display system.</li>
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<li><partinfo>BBa_K2656026</partinfo>: Terminator B0015 adapted to type IIS assembly</li>
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</ul>
  
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AEGIS' lamB display system that we created can be used by future iGEM teams on their MoClo reactions with the desired tag to create level 1 constructs. The only requirement is to adapt the tag sequence to the new scars designed, as this part has been adapted.
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To see the design of this part go to the Design tab.
  
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<partinfo>BBa_K3122001 SequenceAndFeatures</partinfo>
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<h2> How to use it </h2>  
  
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<h2>References</h2>
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The only requirement is to adapt the promotor sequence to the MoClo standard, adding the appropiate restriction sites at the ends.
  
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J. Kim, C. Valencia, R. Liu and W. Lin, "Highly-Efficient Purification of Native Polyhistidine-Tagged Proteins by Multivalent NTA-Modified Magnetic Nanoparticles", Bioconjugate Chemistry, vol. 18, no. 2, pp. 333-341, 2007. Available:10.1021/bc060195l.
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<partinfo>BBa_K3122001 SequenceAndFeatures</partinfo>

Revision as of 02:45, 22 October 2019


6xHis Tag (Adapted to AEGIS' lamB display system)

This part is the constitutive medium promoter, from Anderson family, was adapted to type IIS enzymes standard.

It has been used to express our Level 1 constructs related to membrane proteins. The reason behind this choice relays on its relative strength among other members of the family: it is a medium- strength regulator (0.36) which should not express too much membrane protein as to induce cell lysis.

Characterization

Characterization of this part has been conducted by building two transcriptional units (BBa_K3122004, BBa_ K3122005), and performing Lambda fague and Cell separation assays.

BBa_K3122004 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:

  • BBa_K3122003: Promoter J23107 adapted to type IIS assembly
  • BBa_K2656009: RBS  B0030 adapted to type IIS assembly
  • BBa_K3122002: coding sequence. This composite part is built with BBa_K3122000 (LamB protein) and BBa_K3122001 (6xHis tag) creating our AEGIS' lamB display system exposing 6xHis Tag.
  • BBa_K2656026: Terminator B0015 adapted to type IIS assembly

BBa_K3122005 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:

  • BBa_K3122003: Promoter J23107 adapted to type IIS assembly
  • BBa_K2656009: RBS  B0030 adapted to type IIS assembly
  • BBa_K3122000: LamB coding sequence. This part is our AEGIS' lamB display system.
  • BBa_K2656026: Terminator B0015 adapted to type IIS assembly

To see the design of this part go to the Design tab.

How to use it

The only requirement is to adapt the promotor sequence to the MoClo standard, adding the appropiate restriction sites at the ends.


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
    COMPATIBLE WITH RFC[1000]