Composite

Part:BBa_K3780017

Designed by: Sumohit Sharma   Group: iGEM21_Stuttgart   (2021-09-27)


Dsup N-terminal His tag

The DNA fragment BBa_3780017 contains the information for the Dsup protein. Dsup is a protein that is unique to tardigrades. It protects DNA from UV and ROS damage. Snapgene was used to build in silico the BioBricks and plasmid DSUP construct. The fragments were cloned into the pBAD18 by Gibson Assembly. Two different E. Coli strains were transformed with the fragment (DH5alpha, CW25113) to test the UV protection potential of Dsup in E. Coli. Additionally, the fragment contains an N-terminal His tag, therefore the protein can be purified by chromatography. Therefore the ability of UV protection of Dsup can be tested ex vivo. The results are compared with BBa_K3780013 to validate a possible impact of the his tag.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 1589
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal NheI site found at 203
    Illegal SpeI site found at 1590
    Illegal PstI site found at 1604
    Illegal NotI site found at 7
    Illegal NotI site found at 1597
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal BamHI site found at 142
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 1590
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1
    Illegal XbaI site found at 16
    Illegal SpeI site found at 1590
    Illegal PstI site found at 1604
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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