Device

Part:BBa_K3629017

Designed by: Sravya Kakumanu   Group: iGEM20_Calgary   (2020-10-18)
Revision as of 06:30, 27 October 2020 by Sravyakakumanu (Talk | contribs)


T. reesei EGII expression construct with gibson homology sequences and FLAG tag


Usage and Biology

Design

The native signal peptide from T. reesei was removed so it would not interfere with the fused XRP2 secretion tag native to Y. lipolytica.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 2835
    Illegal EcoRI site found at 2698
    Illegal SpeI site found at 749
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 2698
    Illegal NheI site found at 68
    Illegal NheI site found at 810
    Illegal SpeI site found at 749
    Illegal SpeI site found at 2836
    Illegal PstI site found at 2850
    Illegal NotI site found at 7
    Illegal NotI site found at 2843
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 2698
    Illegal BamHI site found at 2786
    Illegal XhoI site found at 125
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 2836
    Illegal EcoRI site found at 2698
    Illegal SpeI site found at 749
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1
    Illegal EcoRI site found at 2698
    Illegal XbaI site found at 16
    Illegal SpeI site found at 749
    Illegal SpeI site found at 2836
    Illegal PstI site found at 2850
    Illegal NgoMIV site found at 1496
    Illegal NgoMIV site found at 2030
    Illegal NgoMIV site found at 2348
  • 1000
    COMPATIBLE WITH RFC[1000]

There is an EcoRI site within the XRP2 terminator, and an SpeI site within the EXP promoter making this part RFC10 incompatible. However, we added the BioBrick prefix and suffix so that the other enzymes (NotI, XbaI, and PstI) could be used to clone this part into an iGEM plasmid or another plasmid. This part can also be cloned through RFC1000 assembly.


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Categories
Parameters
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