Composite
Part:BBa_K5043008:Design
Designed by: Richard Meuten Group: iGEM24_TUDarmstadt (2024-09-07)
Pyrene degradation operon
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3199
Illegal EcoRI site found at 7727
Illegal PstI site found at 719
Illegal PstI site found at 4703
Illegal PstI site found at 5754 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3199
Illegal EcoRI site found at 7727
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal PstI site found at 719
Illegal PstI site found at 4703
Illegal PstI site found at 5754 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3199
Illegal EcoRI site found at 7727
Illegal BglII site found at 390
Illegal BglII site found at 4695
Illegal BamHI site found at 1259
Illegal BamHI site found at 3644
Illegal XhoI site found at 54
Illegal XhoI site found at 931
Illegal XhoI site found at 1880
Illegal XhoI site found at 2362
Illegal XhoI site found at 3909
Illegal XhoI site found at 8073 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3199
Illegal EcoRI site found at 7727
Illegal PstI site found at 719
Illegal PstI site found at 4703
Illegal PstI site found at 5754 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3199
Illegal EcoRI site found at 7727
Illegal PstI site found at 719
Illegal PstI site found at 4703
Illegal PstI site found at 5754
Illegal NgoMIV site found at 749
Illegal NgoMIV site found at 854
Illegal NgoMIV site found at 1957
Illegal NgoMIV site found at 2637
Illegal NgoMIV site found at 4039
Illegal NgoMIV site found at 4342
Illegal NgoMIV site found at 8108
Illegal AgeI site found at 1350
Illegal AgeI site found at 1408 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Medium expression constitutive promoter J23110 was chosen for this operon. To detect produced proteins, all proteins carry a hexahistidine-tag. His-tags and RBS were added via overhang-PCR. Assembly of operon was carried out successively using GoldenBraid method. [5]
Source
Protein coding sequences are derived from genomes of Mycobacterium vanbaalenii Pyr-1, Nocardioides sp. strain KP7 and Arthrobacter keyseri 12B.