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Part:BBa_K931009

Designed by: Chris Clifford   Group: iGEM12_Nevada   (2012-09-29)

An L-Arabinose Controlled Thiamine Binding - Starch Binding Protein Fusion

A starch-binding protein gene (CBM21) and a thiamine-binding protein gene (ThiT) were placed in sequence with each other in that ThiT comes before CBM21. CBM21 comes from R. oryzae. It is part of an operon that codes for a two-domain enzyme, one of which binds starch molecules. ThiT comes from E.coli and plays a crucial role in the thiamine transport system by binding the thiamine molecule. It has a Kd of 170 nM when the optimum pH is at 5.0 These two genes were placed after an L. Arabinose promoter sequence (BBa_K206000). This allows the protein to be expressed by adding the L. Arabinose chemical. Two terminator sequences (B0010 B0012) were placed at the end of the construct. The resulting gene construct is 2038 base pairs long. The protein complex from these two genes has a starch-binding domain and a thiamine binding domain which can bind their resulting substrates.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 371
    Illegal XbaI site found at 16
    Illegal XbaI site found at 386
    Illegal SpeI site found at 343
    Illegal SpeI site found at 1061
    Illegal PstI site found at 357
    Illegal PstI site found at 1075
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 371
    Illegal SpeI site found at 343
    Illegal SpeI site found at 1061
    Illegal PstI site found at 357
    Illegal PstI site found at 1075
    Illegal NotI site found at 7
    Illegal NotI site found at 350
    Illegal NotI site found at 377
    Illegal NotI site found at 1068
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 371
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 343
    Illegal EcoRI site found at 371
    Illegal XbaI site found at 386
    Illegal SpeI site found at 1061
    Illegal PstI site found at 1075
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1
    Illegal EcoRI site found at 371
    Illegal XbaI site found at 16
    Illegal XbaI site found at 386
    Illegal SpeI site found at 343
    Illegal SpeI site found at 1061
    Illegal PstI site found at 357
    Illegal PstI site found at 1075
  • 1000
    COMPATIBLE WITH RFC[1000]


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