Part:BBa_K4370010

STREPTO_E.coli_CLONING_MODULE

The part allows the easy cloning of Streptomyces genes under the control of a strong and constitutive promoter using a red/white screening. Therefore the module contains parts specific to Streptomyces chassis as well as parts dedicated to the expression in E. coli to perform cloning.

Notably, this part contains the Streptomyces constitutive strong promoter KasOP* followed by a RBS from of capsid protein obtained from Streptomyces temperate phage ϕC31 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). This RFP expression module is followed by two stop codons and two terminators (BB0010 and B0012).

This cassette is meant to be replaced by the construction of interest and offers an easy means of visual screening the clones containing the new construction (pink if the E. coli cassette is present in the plasmid, white if it has been replaced by the gene of interest).

This module is also designed to be easily cloned between NotI sites in the collection of Streptomyces integrative vectors published by Aubry and collaborators (AEM, 2019 - https://journals.asm.org/doi/10.1128/AEM.00485-19).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Unknown
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Unknown
• 1000
  COMPATIBLE WITH RFC[1000]