Generator
KanMX(lox)

Part:BBa_K300989

Designed by: Lorenzo Pasotti, Giacomo Zambianchi, Federica Sampietro, Paolo Magni   Group: iGEM10_UNIPV-Pavia   (2010-10-03)

[LoxP-KanMX-LoxP] excisable marker for yeast

This cassette is to be used in yeast.

It is composed by the kanMX gene from the E. coli transposon Tn903 that confers the resistance to the aminoglycoside antibiotic G418 to yeast (commonly used at 200 ug/ml working concentration). This gene is expressed by the TEF2 promoter/RBS and terminated by the TEF2 transcriptional terminator from Ashbya gossypii filamentous fungus.

The cassette is flanked by two loxP sites and for this reason it can be excised with Cre/loxP mediated marker removal recombination procedure.


ADVANTAGES IN GENOME ENGINEERING APPLICATIONS:

This part can be used to select positive integrants in the yeast genome and then it can be eliminated through Cre/loxP procedure. This enables the engineering of yeast genome in multiple steps, always using the same dominant selection marker.

Sequence and Features

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 39
    Illegal PstI site found at 254
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 254
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 44
    Illegal XhoI site found at 1492
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 39
    Illegal PstI site found at 254
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 39
    Illegal PstI site found at 254
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal XbaI site found at 39
    Illegal PstI site found at 254


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Parameters
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