Difference between revisions of "Part:BBa K2332041:Design"

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===Source===
 
===Source===
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SpyCatcher sequence was obtained from [https://parts.igem.org/Part:BBa_K1159200 BBa_K1159200] and GFP from:
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Intimin protein sequence obtained from: http://www.uniprot.org/uniprot/P43261#sequences And reverse translated using: http://www.bioinformatics.org/sms2/rev_trans.html Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001 SpyCatcher: BBa_K1159200 GFPmut 3b: Part:BBa_E0040
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[https://parts.igem.org/Part:BBa_E0040 BBa_E0040]. Pblind promoter was designed by Jayaraman P. et al. (2016). The DNA sequence was synthesised by Integrated DNA Technologies (IDT)
  
 
===References===
 
===References===
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1. Jayaraman P, Devarajan K, Chua T, Zhang H, Gunawan E, Poh C. Blue light-mediated transcriptional activation and repression of gene expression in bacteria. Nucleic Acids Research. 2016;44(14):6994-7005.
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2. Zakeri B, Fierer J, Celik E, Chittock E, Schwarz-Linek U, Moy V et al. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proceedings of the National Academy of Sciences. 2012;109(12):E690-E697.

Latest revision as of 23:34, 23 October 2017


Blue light inducible expression of GFP-SpyCatcher (Pblind GFP-SpyCatcher)


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.rc site found at 741


Design Notes

We removed the stop codon in GFP and placed the SpyCatcher sequence after a linker sequence and included a HisTag for protein purification. Pblind promoter was designed by Jayaraman P. et al. (2016)

Source

SpyCatcher sequence was obtained from BBa_K1159200 and GFP from: BBa_E0040. Pblind promoter was designed by Jayaraman P. et al. (2016). The DNA sequence was synthesised by Integrated DNA Technologies (IDT)

References

1. Jayaraman P, Devarajan K, Chua T, Zhang H, Gunawan E, Poh C. Blue light-mediated transcriptional activation and repression of gene expression in bacteria. Nucleic Acids Research. 2016;44(14):6994-7005.

2. Zakeri B, Fierer J, Celik E, Chittock E, Schwarz-Linek U, Moy V et al. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proceedings of the National Academy of Sciences. 2012;109(12):E690-E697.