Coding
Part:BBa_K863012:Design
Designed by: Isabel Huber Group: iGEM12_Bielefeld-Germany (2012-09-18)
tthl laccase from T. thermophilus with constitutive promoter J23100, RBS and His-tag
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 37
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1436
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 37
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 37
Illegal NgoMIV site found at 503
Illegal NgoMIV site found at 990 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 818
Design Notes
- PCR on BBa_K863011 with primers with BioBrick Prefix in the fwd primer and Suffix and His-tag in the rev primer.
- fwd: 5'-ACGTGAATTCGCGGCCGCTTCTAGATGCTGGCGCGCAGGAGCTT-3'
- rev: 5'-ACGTCTGCAGCGGCCGCTACTAGTATTATTAGTGATGGTGATGGTGATGACCCACCTCGAGGACTC-3'
- Cloning into BBa_J23100 + BBa_J61101 + pSB1A2.
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.