DNA

Part:BBa_K863202:Design

Designed by: Julia Schirmacher   Group: iGEM12_Bielefeld-Germany   (2012-09-23)

his4 gene from Pichia pastoris


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 2555
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1850
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
    Illegal BamHI site found at 358
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 2555
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 2555
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 510


Design Notes

The DNA sequence from plasmid pPIC9K of Invitrogen (http://products.invitrogen.com/ivgn/product/V17520?ICID==%3Dsearch-product) was used for primer design. But the genomic DNA of Pichia pastoris wildtype X33 was used for amplification via PCR.

Primers for isolation of the sequence with BioBrick Prefix in the fwd primer and Suffix in the rev primer.

fwd: 5'-ACGTgaattcgcggccgcttctagagAGATCTCCTGATGACTGACT-3'

rev: 5'-ctgcagcggccgctactagtaGATCTATCGAATCTAAATGT-3'

Source

Pichia pastoris wild type X-33

References