Difference between revisions of "Part:BBa K2549009"
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<partinfo>BBa_K2549009 short</partinfo>
 
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This part is the N-terminal fragment of Cfa. Cfa is a consensus sequence from an alignment of 73 naturally occurring DnaE inteins that are predicted to have fast splicing rates. Cfa demonstrates both rapid protein splicing and unprecedented thermal and chaotropic durability. Besides, when the Cfa’N is fused to other proteins, it exhibits higher expression levels than other N-terminal fusions. Cfa’N is used in our amplifier to accomplish some complex logic functions. It can also be used by other iGEM teams to assembly their intein-based protein libraries.
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This part is the N-terminal fragment of Cfa. Split intein is a useful protein engineering approach to combine signals<ref>Protein trans-splicing and its use in structural biology: opportunities and limitations. Volkmann G, Iwaï H. Mol Biosyst, 2010 Nov;6(11):2110-21  PMID: 20820635; DOI: 10.1039/c0mb00034e</ref><ref>Cell-Based Biosensors Based on Intein-Mediated Protein Engineering for Detection of Biologically Active Signaling Molecules. Jeon H, Lee E, Kim D, ..., Kim S, Kwon Y. Anal Chem, 2018 Aug;90(16):9779-9786  PMID: 30028129; DOI: 10.1021/acs.analchem.8b01481</ref>. Cfa is a consensus sequence from an alignment of 73 naturally occurring DnaE inteins that are predicted to have fast splicing rates. Cfa demonstrates both rapid protein splicing and unprecedented thermal and chaotropic durability. Besides, when the CfaN is fused to other proteins, it exhibits higher expression levels than other N-terminal fusions<ref>Improved protein splicing using embedded split inteins. Gramespacher JA, Stevens AJ, Thompson RE, Muir TW. Protein Sci, 2018 Mar;27(3):614-619  PMID: 29226478; DOI: 10.1002/pro.3357</ref><ref>Design of a Split Intein with Exceptional Protein Splicing Activity. Stevens AJ, Brown ZZ, Shah NH, ..., Cowburn D, Muir TW. J Am Chem Soc, 2016 Feb;138(7):2162-5  PMID: 26854538; DOI: 10.1021/jacs.5b13528</ref><ref>A promiscuous split intein with expanded protein engineering applications. Stevens AJ, Sekar G, Shah NH, ..., Cowburn D, Muir TW. Proc Natl Acad Sci U S A, 2017 Aug;114(32):8538-8543  PMID: 28739907; DOI: 10.1073/pnas.1701083114</ref>. CfaN is used in our amplifier to accomplish some complex logic functions. It can also be used by other iGEM teams to assembly their intein-based protein libraries.
  
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
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===Usage and Biology===
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TBA
  
  
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===Functional Parameters===
 
===Functional Parameters===
 
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===References===

Revision as of 03:40, 7 October 2018


split intein Cfa N

This part is the N-terminal fragment of Cfa. Split intein is a useful protein engineering approach to combine signals[1][2]. Cfa is a consensus sequence from an alignment of 73 naturally occurring DnaE inteins that are predicted to have fast splicing rates. Cfa demonstrates both rapid protein splicing and unprecedented thermal and chaotropic durability. Besides, when the CfaN is fused to other proteins, it exhibits higher expression levels than other N-terminal fusions[3][4][5]. CfaN is used in our amplifier to accomplish some complex logic functions. It can also be used by other iGEM teams to assembly their intein-based protein libraries.

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
    COMPATIBLE WITH RFC[1000]


Usage and Biology

TBA



References

  1. ↑ Protein trans-splicing and its use in structural biology: opportunities and limitations. Volkmann G, Iwaï H. Mol Biosyst, 2010 Nov;6(11):2110-21 PMID: 20820635; DOI: 10.1039/c0mb00034e
  2. ↑ Cell-Based Biosensors Based on Intein-Mediated Protein Engineering for Detection of Biologically Active Signaling Molecules. Jeon H, Lee E, Kim D, ..., Kim S, Kwon Y. Anal Chem, 2018 Aug;90(16):9779-9786 PMID: 30028129; DOI: 10.1021/acs.analchem.8b01481
  3. ↑ Improved protein splicing using embedded split inteins. Gramespacher JA, Stevens AJ, Thompson RE, Muir TW. Protein Sci, 2018 Mar;27(3):614-619 PMID: 29226478; DOI: 10.1002/pro.3357
  4. ↑ Design of a Split Intein with Exceptional Protein Splicing Activity. Stevens AJ, Brown ZZ, Shah NH, ..., Cowburn D, Muir TW. J Am Chem Soc, 2016 Feb;138(7):2162-5 PMID: 26854538; DOI: 10.1021/jacs.5b13528
  5. ↑ A promiscuous split intein with expanded protein engineering applications. Stevens AJ, Sekar G, Shah NH, ..., Cowburn D, Muir TW. Proc Natl Acad Sci U S A, 2017 Aug;114(32):8538-8543 PMID: 28739907; DOI: 10.1073/pnas.1701083114