Part:BBa_K2549009

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]

Biology

Protein trans-splicing

xxx at al stated: xxxxxx

CfaN and CfaC

xxx at al stated: xxxxxx

References

↑ 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
↑ 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
↑ 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
↑ 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
↑ 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

[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

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