Difference between revisions of "Part:BBa J179002"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J179002 short</partinfo>
 
<partinfo>BBa_J179002 short</partinfo>
  
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the thrC locus and can be selected with Spectinomycin cassette. It has a ampicillin resistance for cloning in E.coli. This backbone is a BioBricked version of the B. subtilis vector pDG1731.  
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This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the thrC locus and can be selected with Spectinomycin cassette. It has a ampicillin resistance for cloning in E.coli. This backbone is a ''BioBricked'' version of the ''B. subtilis'' vector pDG1731.
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This version is derived from [https://parts.igem.org/Part:BBa_K823022 BBa_K823022] and contains neither NgoMIV nor AgeI restriction sites, therefore is RFC25-compatible.
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[[Image:LMU-Munich-PSBBs4S.png|600px]]
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This BioBrick is part of the LMU 2012 igem project [http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks '''''Bacillus''B'''io'''B'''rick'''B'''ox]
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For handling of ''B. subtilis'' vectors, please see [https://static.igem.org/mediawiki/2012/c/c4/LMU-Munich_2012_Bacillus_subtilis_vectors.pdf here].
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The transformation into ''B. subtilis'' is explained [https://static.igem.org/mediawiki/2012/4/41/LMU-Munich_2012_Transformation_of_Bacillus_subtilis.pdf here].
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[[Image:LMU-Munich-Thrplate.png| 300px|left]]
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We tested the integration of this plasmid with the threonine test: The colonies obtained from the transformation are streaked on three plates: LB with 100mg/L spectinomycine, minimal medium with threonine and minimal medium without threonine. As minimal medium, the MNGE-medium described in the transformation protocol was used. The plates are incubated at 37°C over night. Positive colonies grow on LB and minimal medium with threonine, but not on minimal medium without threonine.
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Reference: [http://www.ncbi.nlm.nih.gov/pubmed/8973347 Guérout-Fleury ''et al.'']
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:10, 20 December 2013

pBs4S: Bacillus subtilis vector, thrC integration, spec resistance

This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the thrC locus and can be selected with Spectinomycin cassette. It has a ampicillin resistance for cloning in E.coli. This backbone is a BioBricked version of the B. subtilis vector pDG1731.

This version is derived from BBa_K823022 and contains neither NgoMIV nor AgeI restriction sites, therefore is RFC25-compatible.

LMU-Munich-PSBBs4S.png


This BioBrick is part of the LMU 2012 igem project [http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks BacillusBioBrickBox]

For handling of B. subtilis vectors, please see here.

The transformation into B. subtilis is explained here.

LMU-Munich-Thrplate.png

We tested the integration of this plasmid with the threonine test: The colonies obtained from the transformation are streaked on three plates: LB with 100mg/L spectinomycine, minimal medium with threonine and minimal medium without threonine. As minimal medium, the MNGE-medium described in the transformation protocol was used. The plates are incubated at 37°C over night. Positive colonies grow on LB and minimal medium with threonine, but not on minimal medium without threonine.


Reference: [http://www.ncbi.nlm.nih.gov/pubmed/8973347 Guérout-Fleury et al.]

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4552
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 4558
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4552
    Illegal BamHI site found at 4534
    Illegal XhoI site found at 1011
    Illegal XhoI site found at 1929
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 4552
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 4552
    Plasmid lacks a suffix.
    Illegal XbaI site found at 4567
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2856
    Illegal SapI.rc site found at 605