Plasmid

Part:BBa_M39203

Designed by: Hannah Kempton   Group: MIT BE 20.20-S12   (2012-05-15)


Vector for creation of micro-dystrophin SFV

The general purpose of our part is to help create a modified Semliki Forest Virus that contains micro-dystrophin mRNA in its capsid, which hopefully will be used to deliver said mRNA to muscle cells of muscular dystrophy patients. This DNA vector contains an RNA polymerase promoter, the packaging sequence from SFV (Semliki Forest Virus), the DNA sequence for micro-dystrophin, and an RNA polymerase terminator. Once this vector is transcribed into RNA in vitro, the resultant RNA is introduced into a cell in the lab. A separate helper vector (not included in this part) will be introduced into the same cell as well. This vector will simply contain the genes for the structural proteins of SFV. Once the helper vector is translated to produce the structural capsid proteins, the virus will assemble within the cell, packaging the micro-dystrophin mRNA within the capsid of the virus. The purpose of removing the non-structural genes (other than the necessary packaging sequence) is to remove the replicating and virulent ability of the virus, so that it will not replicate within the body. This virus will then break free of the cell in which it was produced, at which point it can be harvested and injected into patients to treat dystrophin deficiency within muscle cells.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2190
    Illegal PstI site found at 2949
    Illegal PstI site found at 3213
    Illegal PstI site found at 3312
    Illegal PstI site found at 3778
    Illegal PstI site found at 4149
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2190
    Illegal NheI site found at 1337
    Illegal PstI site found at 2949
    Illegal PstI site found at 3213
    Illegal PstI site found at 3312
    Illegal PstI site found at 3778
    Illegal PstI site found at 4149
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2190
    Illegal BglII site found at 1483
    Illegal BglII site found at 1697
    Illegal BglII site found at 2607
    Illegal BamHI site found at 228
    Illegal BamHI site found at 2743
    Illegal BamHI site found at 3448
    Illegal XhoI site found at 2794
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2190
    Illegal PstI site found at 2949
    Illegal PstI site found at 3213
    Illegal PstI site found at 3312
    Illegal PstI site found at 3778
    Illegal PstI site found at 4149
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2190
    Illegal PstI site found at 2949
    Illegal PstI site found at 3213
    Illegal PstI site found at 3312
    Illegal PstI site found at 3778
    Illegal PstI site found at 4149
    Illegal NgoMIV site found at 4329
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2252
    Illegal SapI.rc site found at 2396


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