Difference between revisions of "Part:BBa K2333434"
Line 18: Line 18:
 
<partinfo>BBa_K2333434 parameters</partinfo>
 
<partinfo>BBa_K2333434 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
===References===
 +
 +
[1] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.
 +
 +
[2] 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.

Revision as of 22:27, 29 October 2017


pLac0-1 mf-Lon

This is an IPTG-inducible mf-Lon construct containing the pLlac 0-1 promoter. William and Mary 2017 has modified the mf-Lon gene via codon-optimization for iGEM use and added a double terminator.


Usage and Biology

This composite part is a combination circuit with the LacI repressor under the constitutive promoter J23105 and mf-Lon under the control of the PLlac 0-1 promoter. The mf-Lon protease specifically targets different protein degradation tags with varying affinities corresponding to varying degradation rates. This IPTG-inducible mf-Lon construct was used in tandem with aTc-inducible pdt reporter constructs by William and Mary 2017 to obtain gene expression speed measurements.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 47
    Illegal NheI site found at 70
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 3242
    Illegal AgeI site found at 3326
    Illegal AgeI site found at 3532
    Illegal AgeI site found at 3557
  • 1000
    COMPATIBLE WITH RFC[1000]


References

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

[2] 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.