Coding

Part:BBa_K863010:Design

Designed by: Isabel Huber   Group: iGEM12_Bielefeld-Germany   (2012-09-18)

tthl laccase from Thermus thermophilus with T7 promoter, RBS and His-tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1408
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 475
    Illegal NgoMIV site found at 962
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 790


Design Notes

Primers for isolation of the gene with BioBrick Prefix, pT7 promoter and RBS in the fwd primer and Suffix and His-tag in the rev primer.
fwd: 5'-ACGTGAATTCGCGGCCGCTTCTAGAGtaatacgactcactatagggagagaggagaaaaATGCTGGCGCGCAGGAGCTT-3'
rev: 5'-ACGTCTGCAGCGGCCGCTACTAGTATTATTAGTGATGGTGATGGTGATGACCCACCTCGAGGACTC-3'

Source

The gene sequence was isolated from Thermus thermophilus HB27.

References

Miyazaki, K. (2005). "A hyperthermophilic laccase from Thermus thermophilus HB27." Extremophiles 9(6): 415-425.