Composite

Part:BBa_K3385067

Designed by: Lucas Levassor   Group: iGEM20_DTU-Denmark   (2020-10-16)


T1G3R_Spower_glaA

About the part: Insert of glaA expression cassette for A. niger. It is made to be integrated in albA, which will lead to white/yellow conidia upon correct integration in the genome. The insert is inserted in pSB1C3 via USER cloning to create the integration plasmid for A. niger. The signal peptide was changed to SigPower using USER cloning and primer extension. The integration platform is made from part BBa_K3046030 pGIA2P1 designed by the DTU iGEM 2019 team.

Functionality: This signal peptide was investigated in Deeploc [1] to predict the localization of the protein which it is inserted upstream of.

Localization predicted by Deeploc.


Result: A tissue PCR was performed to show that the synthetic signal peptide can be successfully integrated in the A. niger genome.
Tissue PCR showing the integration of the synthetic signal peptides SigPower, SigPeanut and SigPilot into the A. niger genome.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1858
    Illegal EcoRI site found at 5765
    Illegal EcoRI site found at 7193
    Illegal XbaI site found at 1623
    Illegal XbaI site found at 5796
    Illegal SpeI site found at 1097
    Illegal PstI site found at 893
    Illegal PstI site found at 4027
    Illegal PstI site found at 6603
    Illegal PstI site found at 8337
    Illegal PstI site found at 8938
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1858
    Illegal EcoRI site found at 5765
    Illegal EcoRI site found at 7193
    Illegal NheI site found at 4904
    Illegal SpeI site found at 1097
    Illegal PstI site found at 893
    Illegal PstI site found at 4027
    Illegal PstI site found at 6603
    Illegal PstI site found at 8337
    Illegal PstI site found at 8938
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1858
    Illegal EcoRI site found at 5765
    Illegal EcoRI site found at 7193
    Illegal BglII site found at 3882
    Illegal BglII site found at 5404
    Illegal BglII site found at 8435
    Illegal BamHI site found at 1729
    Illegal BamHI site found at 2157
    Illegal BamHI site found at 4862
    Illegal BamHI site found at 5664
    Illegal BamHI site found at 5790
    Illegal BamHI site found at 6341
    Illegal XhoI site found at 548
    Illegal XhoI site found at 735
    Illegal XhoI site found at 963
    Illegal XhoI site found at 6201
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1858
    Illegal EcoRI site found at 5765
    Illegal EcoRI site found at 7193
    Illegal XbaI site found at 1623
    Illegal XbaI site found at 5796
    Illegal SpeI site found at 1097
    Illegal PstI site found at 893
    Illegal PstI site found at 4027
    Illegal PstI site found at 6603
    Illegal PstI site found at 8337
    Illegal PstI site found at 8938
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1858
    Illegal EcoRI site found at 5765
    Illegal EcoRI site found at 7193
    Illegal XbaI site found at 1623
    Illegal XbaI site found at 5796
    Illegal SpeI site found at 1097
    Illegal PstI site found at 893
    Illegal PstI site found at 4027
    Illegal PstI site found at 6603
    Illegal PstI site found at 8337
    Illegal PstI site found at 8938
    Illegal NgoMIV site found at 2566
    Illegal NgoMIV site found at 5122
    Illegal NgoMIV site found at 8325
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3410
    Illegal BsaI site found at 3548
    Illegal BsaI site found at 4237
    Illegal BsaI.rc site found at 4730
    Illegal SapI site found at 8826
    Illegal SapI site found at 9135


References:
[1] Jose Juan Almagro Armenteros, Casper Kaae Sønderby, Søren Kaae Sønderby, Henrik Nielsen, Ole Winther; DeepLoc: prediction of protein subcellular localization using deep learning, Bioinformatics, btx431

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