Part:BBa_K4370010

KasOP*_promoter_STREPTO The part allows the easy cloning of Streptomyces <i/> genes under the control of a strong and constitutive promoter using a red/white screening of the clones of interest in <i> E. coli <i/> used as a chassis for cloning. Therefore the module contains parts specific to <i> Streptomyces <i/> chassis as well as parts dedicated to the expression in <i> E. coli <i/> to perform cloning.

Notably, this part contains the <i> Streptomyces <i/> constitutive strong promoter KasOP* followed by a RBS from of capsid protein obtained from <i> 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 NdeI site (catATG) allows the easy cloning of genes between NdeI (ATG/start codon) and SpeI sites (this latter site been in the suffixe). Between these two sites is inserted an cassette for the expression of mRFP1 (BBa_E1010) under the control of an E. coli promoter <i/>.

Therefore this module allows the easy red/white cloning of sequences of interest in place of the <i> E. coli <i/> expression module. This part is designed for the cloning of Streptomyces genes of interest in place of this cassette using an easy visual screening (pink if the <i> E. coli <i/> cassette is present in the plasmid, white if it has been replaced by the gene of interest).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 <i> Streptomyces <i/> integrative vectors published by Aubry and collaborators (AEM, 2019).

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]