Coding

Part:BBa_K1351023

Designed by: Patrick Reith   Group: iGEM14_LMU-Munich   (2014-10-04)
Revision as of 17:02, 16 October 2014 by Jara (Talk | contribs)

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.


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


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Parameters
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