Plasmid_Backbone

Part:BBa_K245008:Design

Designed by: Nika Debeljak   Group: iGEM09_Slovenia   (2009-09-26)


BB-NIC-III-HisN


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2119
    Illegal XbaI site found at 2244
    Illegal SpeI site found at 13
    Illegal PstI site found at 77
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2119
    Illegal SpeI site found at 13
    Illegal PstI site found at 77
    Illegal NotI site found at 70
    Illegal NotI site found at 2125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2119
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2119
    Illegal XbaI site found at 2244
    Illegal SpeI site found at 13
    Illegal PstI site found at 77
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2119
    Illegal XbaI site found at 2244
    Illegal SpeI site found at 13
    Illegal PstI site found at 77
    Illegal NgoMIV site found at 2250
    Illegal AgeI site found at 1
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 1158


Design Notes

ccdB was multiplied from BB-NIC-II-HisN (BBa_K245004) by PCR using primers which contained restriction sites in different combinations than those found in BB-NIC-II-HisN next to ccdB. Fragment was then ligated in BB-NIC-II-HisN cut with NgoMIV and SpeI.


Source

The basis for BB-NIC-III-HisN is vector pSB1A2. Modified prefix and suffix are of synthetic origin. ccdB originates from part BBa_K245004.

References