Plasmid

Part:BBa_K802003:Design

Designed by: Sandu Ioana   Group: iGEM12_Lyon-INSA   (2012-09-20)
Revision as of 12:27, 21 September 2012 by Mwolfovski (Talk | contribs) (→‎Design Notes)

Shuttle vector for E. coli and B. subtilis


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 6506
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6506
    Illegal NheI site found at 3377
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 6512
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6506
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 6506
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 6506
    Illegal XbaI site found at 6521
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3203
    Illegal BsaI.rc site found at 5190


Design Notes

The SpeI site from the B. subtilis coding region of the pHT304 plasmid was eliminated by filling-in. Afterwards, an iGEM linker containing all 4 of the restriction enzyme sites (EcoRI,XbaI,SpeI,PstI) was cloned into the modified vector.

L35.png

Source

The shuttle vector is derived from pHT304 (Arantes & Lereclus, 1991). It is a low copy plasmid number in B. subtilis.

References