Difference between revisions of "Part:BBa K2333402:Design"

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===Design Notes===
 
===Design Notes===
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This part was designed to include both a double stop codon and double terminator after the pdt tag, which enables it to be appended to an arbitrary protein in a given circuit, without changing the underlying architecture.
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This part was designed to include both a double stop codon and double terminator after the pdt, which enables it to be appended to an arbitrary protein in a given circuit, without changing the underlying architecture.
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===Source===
 
===Source===
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Pdt B was originally generated by mutagenesis from the endogenous Lon degraded tags from the bacteria Mycoplasma florum by Collins et al. 2014 "Tunable Protein Degradation in Bacteria". Pdt B corresponds to Collins et al.'s tag pdt#3a. To create pdt B, the amino acid sequence was taken from Collins et al. and was codon optimized for E. coli, then synthesized by IDT.
  
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The tag pdt #3 is originally generated by mutagenesis from the endogenous Lon degraded tags from the bacteria Mycoplasma florum by Collins et al. 2014 "Tunable Protein Degradation in Bacteria". We synthesized the tag by IDT and cloned using gibson assembly.
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UNS sequences are from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly.
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B0015 comes from the registry, and the UNS sequence comes from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly.
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UNS 2 Sequence is a unique oligonucleotide sequence introduced by William and Mary iGEM 2016 . See BBa_K2066018 and BBa_K2066019.
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===References===
 
===References===
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[2] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.
 
[2] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.
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[3] Part:BBa_K2066018. Part:BBa K2066018. [accessed 2017 Jun 16]. https://parts.igem.org/Part:BBa_K2066018
 
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[4] Part:BBa_K2066018. Part:BBa K2066019. [accessed 2017 Jun 16]. https://parts.igem.org/Part:BBa_K2066019
 

Latest revision as of 02:07, 29 October 2017


Cloning ready protein degradation tag B (medium-strong) with double terminator


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 41
    Illegal BsaI.rc site found at 263


Design Notes

This part was designed to include both a double stop codon and double terminator after the pdt, which enables it to be appended to an arbitrary protein in a given circuit, without changing the underlying architecture.

Source

Pdt B was originally generated by mutagenesis from the endogenous Lon degraded tags from the bacteria Mycoplasma florum by Collins et al. 2014 "Tunable Protein Degradation in Bacteria". Pdt B corresponds to Collins et al.'s tag pdt#3a. To create pdt B, the amino acid sequence was taken from Collins et al. and was codon optimized for E. coli, then synthesized by IDT.

UNS sequences are from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly.

References

[1] Torella JP, Boehm CR, Lienert F, Chen J-H, Way JC, Silver PA. Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic Acids Research. 2013;42(1):681–689.

[2] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.