DNA

Part:BBa_K3046033:Design

Designed by: Philip Srensen   Group: iGEM19_DTU-Denmark   (2019-10-21)


IS2 downstream sequence for A. niger genome integration


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 118
    Illegal PstI site found at 1262
    Illegal PstI site found at 1863
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 118
    Illegal PstI site found at 1262
    Illegal PstI site found at 1863
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 118
    Illegal BglII site found at 1360
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 118
    Illegal PstI site found at 1262
    Illegal PstI site found at 1863
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 118
    Illegal PstI site found at 1262
    Illegal PstI site found at 1863
    Illegal NgoMIV site found at 1250
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1751
    Illegal SapI site found at 2060


Design Notes

This part is not RFC[1000] compatible, and is therefore best used with cloning methods that does not rely on either 3A or TypeIIS assembly. DTU-Denmark 2019 has used Gibson assembly for this part with great success. It should be possible to de novo synthesize this part. Generally the part does not contain too many repeats, and we have had no problems with designing primers for PCR/sequencing that binds in this sequence.


Source

Our IS2 parts were created by Dorthe Holm in 2013 as part of a ph.d. thesis [1]

References

[1] Holm, D. K. (2013). Development and Implementation of Novel Genetic Tools for Investigation of Fungal Secondary Metabolism. Kgs. Lyngby: Technical University of Denmark.