Coding

Part:BBa_K863000:Design

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

bpul (laccase from Bacillus pumilus) 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 BglII site found at 156
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 523
    Illegal NgoMIV site found at 991
    Illegal AgeI site found at 279
    Illegal AgeI site found at 1156
  • 1000
    COMPATIBLE WITH RFC[1000]


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'-ACGTGAATTCGCGGCCGCTTCTAGAGtaatacgactcactatagggagagaggagaaaaATGAACCTAGAAAAATTTGT-3'
rev: 5'-ACGTCTGCAGCGGCCGCTACTAGTATTATTAGTGATGGTGATGGTGATGCTGGATGATATCCATCG-3'

Source

The gene sequence was originally isolated from Bacillus pumilus DSM 27 (ATCC7061).

References

Reiss, R., J. Ihssen, et al. (2011). "Bacillus pumilus laccase: a heat stable enzyme with a wide substrate spectrum." BMC Biotechnol 11: 9.