Difference between revisions of "Part:BBa K1351023"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1351023 short</partinfo>
 
<partinfo>BBa_K1351023 short</partinfo>
  
Catalyzes the conversion of uracil and 5-phospho-alpha-D-ribose 1-diphosphate (PRPP) to UMP and diphosphate
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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.
Will allow negative selection, Will be Implementet in Modified Plasmid Backbones pSB2E, pSB4S, pSB1C to allow resistance free cloning
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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.
  
 
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