Plasmid_Backbone

Part:BBa_K245007:Design

Designed by: Nika Debeljak   Group: iGEM09_Slovenia   (2009-09-25)


BB-NIC-II-HisC


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2082
    Illegal XbaI site found at 2150
    Illegal SpeI site found at 13
    Illegal PstI site found at 40
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2082
    Illegal SpeI site found at 13
    Illegal PstI site found at 40
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2082
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2082
    Illegal XbaI site found at 2150
    Illegal SpeI site found at 13
    Illegal PstI site found at 40
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2082
    Illegal XbaI site found at 2150
    Illegal SpeI site found at 13
    Illegal PstI site found at 40
    Illegal NgoMIV site found at 2156
    Illegal AgeI site found at 2162
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 1121


Design Notes

Using overlapping primers and PCR ligation a fragment containing modified prefix, ccdB domain and modified suffix with His tag inserted between multiple-cloning site and stop codon was first obtained. The fragment was then ligated in pSB1A2 vector cut with EcoRI and PstI


Source

The basis for BB-NIC-II-HisC is vector pSB1A2. Modified prefix and suffix are of synthetic origin. ccdB originates from part BBa_P1010.

References