Translational_Unit

Part:BBa_K317041:Design

Designed by: Yoshiyuki MURAKAMI   Group: iGEM10_Tokyo-NoKoGen   (2010-10-21)

RBS(B0034)-pduABJKNU-Term(B0015)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1692
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2476
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

①We used following primers to clone each gene

pduAB cloning forward : 5'-TATGAATTCTCTAGATGCAACAAGAAGCG-3'

pduAB cloning reverse : 5'-TATACTAGTTCAGATGTAGGACGGACG-3'

pduJK cloning forward : 5'-TCTGAATTCTCTAGATGAAGAACGCACTG-3'

pduJK cloning reverse : 5'-ATTACTAGTTCACGCTTCACCTCG-3'

pduN cloning forward : 5'-TATGAATTCTCTAGATGCATCTGGCACGG-3'

pduN cloning reverse : 5'-TAGACTAGTCTAACGAGAAAGCGTGTCGA-3'

pduU cloning forward : 5'-TCTGAATTCTCTAGATGGAAAGACAACCC-3'

pduU cloning reverse : 5'-TCTACTAGTTTATGTCCGGGTGATGGG-3'

②We used following primers to modify Pst1 sites in pduAB and pduJK

pduA pst1 mofify forward 5'-GGCTTGACTGCGGCCATAGAGG-3'

pduA pst1 mofify reverse 5'-CCTCTATGGCCGCAGTCAAGCC-3'

pduB pst1 modify forward 5'-CAGAGAAAAGCTGTAGTTTAACGG-3'

pduB pst1 modify reverse 5'-CCGTTAAACTACAGCTTTTCTCTG-3'

pduJK pst1 modify forward 5'-GCAGGAAGCGCTGCGGCAAGCGCCG-3'

pduJK pst1 modify reverse 5-CGGCGCTTGCCGCAGCGCTTCCTGC-3'

③We used following primers to attach RBS (B0034) to each gene

RBS pduAB forward 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGCAACAAGAAGCG-3'

RBS pduAB reverse 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTATCAGATGTAGGACGGACG-3'

RBS pduJK forward 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGAAGAACGCACTG-3'

RBS pduJK reverse 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTATCACGCTTCACCTCG-3'

RBS pduN forward 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGCATCTGGCACGG-3'

RBS pduN reverse 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTAACGAGAAAGCGTGTCGA-3'

RBS pduU forward 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGAAAGACAACCC-3'

RBS pduU reverse 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTATTATGTCCGGGTGATGGG-3'

Source

Citrobacter freundii

References