Composite

Part:BBa_K4370010:Design

Designed by: Stéphanie Bury-Moné   Group: iGEM22_GO_Paris-Saclay   (2022-08-17)


STREPTO_E.coli_CLONING_MODULE


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 792
    Illegal AgeI site found at 904
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The NdeI site (catATG) allows the easy cloning of genes between NdeI (ATG/start codon) and SpeI sites (this latter site been in the suffixe). This module is also designed to be easily cloned between NotI sites in the collection of Streptomyces integrative vectors published by Aubry and collaborators (Appl Environ Microbiol, 2019).


This part contains the Streptomyces constitutive strong promoter KasOP* (BBa_K4370006) followed by a RBS from of capsid protein obtained from Streptomyces temperate phage ϕC31 (BBa_K4370007) that has been previously described as highly efficient to promote translation when combined with the kasOP* promoter (Bai et al., PNAS, 2015). The part also contains a cassette for the expression of mRFP1 (BBa_K4370008) under the control of an E. coli promoter (BBa_K4370009) and an RBS (BBa_B0034). This RFP expression module is followed by two stop codons and two terminators (BBa_B0010 and BBa_B0012).

Source

The sequence has been synthetized.

References

Chaoxian Bai , Yang Zhang , Xuejin Zhao, Yiling Hu, Sihai Xiang, Jin Miao, Chunbo Lou, Lixin Zhang “Exploiting a precise design of universal synthetic modular regulatory elements to unlock the microbial natural products in Streptomyces”, Proc Natl Acad Sci U S A , 2015 Sep 29;112(39):12181-6. https://www.pnas.org/doi/10.1073/pnas.1511027112

Weishan Wang, Xiao Li, Juan Wang, Sihai Xiang, Xiaozhou Feng, Keqian Yang, “An engineered strong promoter for Streptomycetes”, Appl Environ Microbiol., 2013 Jul;79(14):4484-92. https://doi.org/10.1128/AEM.00985-13

Céline Aubry, Jean-Luc Pernodet , Sylvie Lautru, « Modular and Integrative Vectors for Synthetic Biology Applications in Streptomyces spp”, Appl Environ Microbiol, 2019 Aug 1;85(16):e00485-19 https://doi.org/10.1128/AEM.00485-19