Difference between revisions of "Part:BBa K1351023"
 
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UPP catalyzes the conversion of uracil and 5-phospho-alpha-D-ribose 1-diphosphate (PRPP) to UMP and diphosphate whach are toxic to the cells.
 
UPP catalyzes the conversion of uracil and 5-phospho-alpha-D-ribose 1-diphosphate (PRPP) to UMP and diphosphate whach are toxic to the cells.
 
The cassette will allow negative selection in an upp-deficent ''Bacillus subtilis'' strain.
 
The cassette will allow negative selection in an upp-deficent ''Bacillus subtilis'' strain.
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We plan to implement upp in modified Plasmid Backbones  of [https://parts.igem.org/Part:BBa_J179001 pSB2E], [ https://parts.igem.org/Part:BBa_J179002 pSB4S], [https://parts.igem.org/Part:BBa_J179000 pSB1C] to allow resistance free cloning.
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We plan to implement upp in modified Plasmid Backbones  of [https://parts.igem.org/Part:BBa_J179001 pSB2E], [https://parts.igem.org/Part:BBa_J179002 pSB4S], [https://parts.igem.org/Part:BBa_J179000 pSB1C] to allow resistance free cloning.
  
 
This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:  
 
This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:  
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<span style="color:blue">EcoRI</span> and <span style="color:blue">PstI</span> in blue, <span style="color:green">NotI</span> in green, <span style="color:red">XbaI</span> and <span style="color:red">SpeI</span> in red, <span style="color:orange">NgoMIV</span> and <span style="color:orange">AgeI</span> in orange. Shine-Dalgarno sequence and stop codons are underlined.
 
<span style="color:blue">EcoRI</span> and <span style="color:blue">PstI</span> in blue, <span style="color:green">NotI</span> in green, <span style="color:red">XbaI</span> and <span style="color:red">SpeI</span> in red, <span style="color:orange">NgoMIV</span> and <span style="color:orange">AgeI</span> in orange. Shine-Dalgarno sequence and stop codons are underlined.
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This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus].
  
  

Latest revision as of 20:38, 17 October 2014

upp cassette as counter-selection marker

UPP catalyzes the conversion of uracil and 5-phospho-alpha-D-ribose 1-diphosphate (PRPP) to UMP and diphosphate whach are toxic to the cells. The cassette will allow negative selection in an upp-deficent Bacillus subtilis strain. We plan to implement upp in modified Plasmid Backbones of pSB2E, pSB4S, pSB1C to allow resistance free cloning.

This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:

prefix with EcoRI, NotI, XbaI, SD and NgoMIV: GAATTCGCGGCCGCTTCTAGAG
suffix with AgeI, SpeI, NotI and PstI: ACCGGTTAATACTAGTAGCGGCCGCCTGCAG

Sites of restriction enzymes generating compatible overhangs have the same color:

EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.

This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus].


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 688
    Illegal SapI.rc site found at 466