Difference between revisions of "Part:BBa K5043008:Design"

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===Design Notes===
 
===Design Notes===
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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. [2]
+
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===
 
===Source===

Latest revision as of 13:12, 12 September 2024


Pyrene degradation operon


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE 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
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE 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.

References