Difference between revisions of "Part:BBa K2549010:Design"

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===Design Notes===
 
===Design Notes===
 
This part is the C-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<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>. The 122-124 residues of Cfa is mutated from '''EKD''' to '''GEP''', which has been proved to imbue ultrafast DnaE split inteins with minimal extein dependence, thus improving split Intein-mediated protein cyclization<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>.
 
This part is the C-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<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>. The 122-124 residues of Cfa is mutated from '''EKD''' to '''GEP''', which has been proved to imbue ultrafast DnaE split inteins with minimal extein dependence, thus improving split Intein-mediated protein cyclization<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>.
 +
  
 
===Source===
 
===Source===
 
From IDT (gBlock), codon optimized for human
 
From IDT (gBlock), codon optimized for human
 +
  
 
===References===
 
===References===

Latest revision as of 11:03, 17 October 2018


split intein Cfa C


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 BsaI site found at 91
    Illegal SapI.rc site found at 21


Design Notes

This part is the C-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[3][4][5]. The 122-124 residues of Cfa is mutated from EKD to GEP, which has been proved to imbue ultrafast DnaE split inteins with minimal extein dependence, thus improving split Intein-mediated protein cyclization[6].


Source

From IDT (gBlock), codon optimized for human


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
  6. ↑ 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