Plasmid

Part:BBa_I745003:Design

Designed by: Princeton iGEM2007   Group: iGEM07_Princeton   (2007-10-26)


p148D64V


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1719
    Illegal EcoRI site found at 6093
    Illegal XbaI site found at 1623
    Illegal XbaI site found at 7706
    Illegal SpeI site found at 18
    Illegal SpeI site found at 2497
    Illegal PstI site found at 2405
    Illegal PstI site found at 3829
    Illegal PstI site found at 8179
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1719
    Illegal EcoRI site found at 6093
    Illegal NheI site found at 7718
    Illegal SpeI site found at 18
    Illegal SpeI site found at 2497
    Illegal PstI site found at 2405
    Illegal PstI site found at 3829
    Illegal PstI site found at 8179
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1719
    Illegal EcoRI site found at 6093
    Illegal BglII site found at 3086
    Illegal BglII site found at 6977
    Illegal BglII site found at 7724
    Illegal BamHI site found at 7366
    Illegal BamHI site found at 8373
    Illegal BamHI site found at 8710
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1719
    Illegal EcoRI site found at 6093
    Illegal XbaI site found at 1623
    Illegal XbaI site found at 7706
    Illegal SpeI site found at 18
    Illegal SpeI site found at 2497
    Illegal PstI site found at 2405
    Illegal PstI site found at 3829
    Illegal PstI site found at 8179
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1719
    Illegal EcoRI site found at 6093
    Illegal XbaI site found at 1623
    Illegal XbaI site found at 7706
    Illegal SpeI site found at 18
    Illegal SpeI site found at 2497
    Illegal PstI site found at 2405
    Illegal PstI site found at 3829
    Illegal PstI site found at 8179
    Illegal NgoMIV site found at 1311
    Illegal NgoMIV site found at 1468
    Illegal NgoMIV site found at 1727
    Illegal AgeI site found at 4475
    Illegal AgeI site found at 7697
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2386
    Illegal BsaI.rc site found at 9852
    Illegal SapI site found at 8769
    Illegal SapI.rc site found at 2095
    Illegal SapI.rc site found at 6346


Design Notes

We attempted to mutate the integrase in two ways: two-step PCR and site-directed mutagenesis. The site-directed approach was the best approach.


Source

This is a known helper plasmid for lentiviral infection, modified to be non-integrating.

References