Plasmid_Backbone

Part:BBa_K4501022:Design

Designed by: Pau Marín Escudero   Group: iGEM22_Barcelona_UB   (2022-09-22)


pJUMP lentiviral transfer plasmid


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 5061
    Illegal suffix found at 5105
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5061
    Illegal SpeI site found at 5106
    Illegal PstI site found at 5120
    Illegal NotI site found at 5067
    Illegal NotI site found at 5113
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5061
    Illegal BglII site found at 33
    Illegal BglII site found at 74
    Illegal BglII site found at 3682
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 5061
    Illegal suffix found at 5106
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 5061
    Plasmid lacks a suffix.
    Illegal XbaI site found at 5076
    Illegal SpeI site found at 5106
    Illegal PstI site found at 5120
    Illegal NgoMIV site found at 1895
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal SapI.rc site found at 2693
    Illegal SapI.rc site found at 4194


Design Notes

We firstly designed both 5' and 3' modules to be assembled sequentially, since they contained BsmBI and BsaI restriction sites. However, at the end we decided to re-design the construction and we substituted the parts containing those recognition sites by other versions (from popular lentiviral transfer plasmids of Addgene) that do not contain the sequences. With that, we end up having a device that could be pre-assembled and afterwards main assembly could be performed with no limitation.


Source

We took the components from widespread used trasfer plasmid found on Addgene and Vector Builder: https://www.addgene.org/browse/sequence/241925/ https://www.addgene.org/browse/sequence/237918/ https://en.vectorbuilder.com/resources/vector-system/pLV_Exp.html


References

https://www.addgene.org/browse/sequence/241925/ https://www.addgene.org/browse/sequence/237918/ https://en.vectorbuilder.com/resources/vector-system/pLV_Exp.html