Difference between revisions of "Part:BBa K4370010"
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<partinfo>BBa_K4370010 short</partinfo> | <partinfo>BBa_K4370010 short</partinfo> | ||
− | The part allows the easy cloning of <i> Streptomyces </i> genes under the control of a strong and constitutive promoter using a red/white screening | + | The part allows the easy cloning of <i> Streptomyces </i> genes under the control of a strong and constitutive promoter using a red/white screening. 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 </i> 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 ( | + | Notably, this part contains the <i> Streptomyces </i> constitutive strong promoter KasOP* followed by a RBS from of capsid protein obtained from <i> Streptomyces </i> 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_K4370008) under the control of an <i> E. coli </i> 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 <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 cassette is present in the plasmid, white if it has been replaced by the gene of interest).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). | + | 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 - https://journals.asm.org/doi/10.1128/AEM.00485-19). |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 13:33, 18 August 2022
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 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_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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Unknown
- 1000COMPATIBLE WITH RFC[1000]