Regulatory
pLacUV5e

Part:BBa_K4451000

Designed by: Brooks J Rady   Group: iGEM22_Sheffield   (2022-09-30)


pLacUV5e (lacO1-minus35cons-minus10cons-lacOsym)

BBa_K4451000 (COMBO-lacO1-minus35cons-minus10cons-lacOsym) is a synthetic IPTG-inducible promoter, designed by Yu et al. (2021) as part of a library of 1600 lacUV5 variants (‘Pcombo’) which aimed to elucidate the combinatorial effects of RNA polymerase and operator site strengths on overall gene expression. BBa_K4451000 possesses consensus -35 and -10 RNAP binding sites, which are flanked by LacI repressor binding sites. The synthetic ‘lacOsym’ operator (Spronk et al., 1999) is found proximal to the transcription start site, whereas the second, comparatively weaker O1 operator is found further upstream. This combination of regulatory elements was found to result in a high fold-change between uninduced and induced states, and relatively low leakiness when uninduced.


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]


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Categories
Parameters
None