Plasmid_Backbone
K592200

Part:BBa_K592200:Design

Designed by: Erik Gullberg, Antonio Ascue Avalos, Erik Lundin   Group: iGEM11_Uppsala-Sweden   (2011-09-21)
Revision as of 17:10, 21 September 2011 by Antonio (Talk | contribs)

Low to medium copy BioBrick standard vector


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3116
    Illegal NheI site found at 1403
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3122
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3116
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3116
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3116
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3131
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 1969
    Illegal AgeI site found at 994
    Illegal AgeI site found at 1317
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 289


Design Notes

To make the plasmid usable for Lambda Red recombineering (Datsenko and Wanner, 2000), all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.

Source

The P22 late terminator come from the genome of the Salmonella phage P22 (GenBank: AF217253.1, 19662..19702).

References

Datsenko, K. A. and B. L. Wanner (2000). "One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products." Proc Natl Acad Sci U S A 97(12): 6640-5.