Device

Part:BBa_K2170000:Design

Designed by: Max Mustermann   Group: iGEM16_LMU-TUM_Munich   (2016-10-11)


Biotinylated receptor with biotin acceptor peptide combined with a secretory BirA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 576
    Illegal BglII site found at 3529
    Illegal BamHI site found at 2597
    Illegal XhoI site found at 761
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2372
    Illegal BsaI site found at 4077
    Illegal BsaI.rc site found at 743
    Illegal SapI site found at 2529


Design Notes

Related BioBrick:

  • Related BioBricks:

BBa_K2170002: Secretory eukaryotic biotin binding receptor with single chain avidin
BBa_K2170001: Secretory eukaryotic biotin binding receptor with enhanced monomeric avidin


Cloning details:

  • Designed in RFC25

Quality control measures:

  • Test digestion using EcorRI & PstI
  • Sequencing using VF2
  • Part was partly sequenced

Backbone:

  • Backbone name: pSB1C3
  • Resistance: Cp
  • Copynumber: high

Protein coding:

  • Protein:

Biotinylated receptor with biotin acceptor peptid [Nucleotide 633 to 2204]
Secretory BirA [Nucleotide 2605 to 3672]

  • Tag: internal A3C5 tag/ C terminal Strep TagII


Enzymatic activity:

  • none

Cytotoxicity:

  • not known

Safety notes:

  • Known and anticipated sefety issues: none
  • Known and anticipated security issues: none

Intellectual property:

Corresponding part author/authors:

Source

Source:

  • Parts synthesized by IDT


Organism:
Genesequence designed for:

  • eucaryotic cells e.g. Mexi or Trex


References

Literature references:
1. [http://www.nature.com/nmeth/journal/v2/n2/full/nmeth735.html Chen, I., Howarth, M., Lin, W., & Ting, A. Y. (2005). Site-specific labeling of cell surface proteins with biophysical probes using biotin ligase. Nature methods, 2(2), 99-104.]
2. [http://jcb.rupress.org/content/108/2/229.short Kozak, M. (1989). The scanning model for translation: an update. The Journal of cell biology, 108(2), 229-241.]
3. [http://nar.oxfordjournals.org/content/15/20/8125.short Kozak, M. (1987). An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic acids research, 15(20), 8125-8148.]
4. [http://www.nature.com/nmeth/journal/v8/n10/abs/nmeth.1701.html Petersen, T. N., Brunak, S., von Heijne, G., & Nielsen, H. (2011). SignalP 4.0: discriminating signal peptides from transmembrane regions. Nature methods, 8(10), 785-786.]
5. [http://content.iospress.com/articles/human-antibodies/hab5-1-2-11 Alexander, H., Harpprecht, J., Podzuweit, H. G., Rautenberg, P., & Müller-Ruchholtz, W. (1994). Human monoclonal antibodies recognize early and late viral proteins of human cytomegalovirus. Human Antibodies, 5(1-2), 81-90. 6. ]
7. [http://journals.plos.org/plosone/article?id=10.1371/journal.pone.000439 Kredel, S., Oswald, F., Nienhaus, K., Deuschle, K., Röcker, C., Wolff, M., ... & Wiedenmann, J. (2009). mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures. PloS one, 4(2), e4391.]
8. Bajar, B. T., Wang, E. S., Lam, A. J., Kim, B. B., Jacobs, C. L., Howe, E. S., ... & Chu, J. (2016). Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting. Scientific reports, 6.
9. [http://www.nature.com/nprot/journal/v2/n6/abs/nprot.2007.209.html Schmidt, T. G., & Skerra, A. (2007). The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins. Nature protocols, 2(6), 1528-1535.]