Coding
oprF

Part:BBa_K1172501:Design

Designed by: Lukas Rositzka   Group: iGEM13_Bielefeld-Germany   (2013-08-28)

Outer membrane porin OprF from Pseudomonas fluorescens


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 764
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1156
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 677


Design Notes

Amplification with Gibson-primer:

  • Amplification of the outer membrane protein (oprF):
Forward primer oprF (49 bp)

5'-GAATTCGCGGCCGCTTCTAGATGAAACTGAAAAACACCTTGGGCTTTGC-3'

Reverse primer oprF (51 bp)

5'-CTGCAGCGGCCGCTACTAGTATTACTTAGCTTGGGCTTCAACCTGCGCTTC-3'
  • Amplification of the pSB1C3-Backbone:
Forward primer pSB1C3 oprF (43 bp)

5'-TTGAAGCCCAAGCTAAGTAATACTAGTAGCGGCCGCTGCAGTC-3'

Reverse primer pSB1C3 oprF (43 bp)

5'-AAGGTGTTTTTCAGTTTCATCTAGAAGCGGCCGCGAATTCCAG-3'


Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].


Source

Pseudomonas fluorescens Migula 1895 genomic DNA ([http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation Whole Genome Isolation]) DSMZ No.: 7355

References