Plasmid_Backbone
K592202

Part:BBa_K592202

Designed by: Antonio Ascue Avalos   Group: iGEM11_Uppsala-Sweden   (2011-09-21)
Revision as of 16:25, 21 September 2011 by Antonio (Talk | contribs)

Low to medium copy Lambda Red recombineering compatible plasmid

BBa_K592202 is a BioBrick friendly vector with low to medium copy p15A replication origin (BBa_I50032) and a kanamycin antibiotic resistance marker, usable for Lambda Red recombineering in E coli. The E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22, and the kanamycin resistance marker is flanked by FRT sites so that the resistance marker can be removed after chromosomal integration using the FLP recombinase.


K592202-J04450.png


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.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1806
    Illegal XbaI site found at 3199
    Illegal PstI site found at 2629
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3409
    Illegal NheI site found at 1403
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal PstI site found at 2629
    Illegal NotI site found at 9
    Illegal NotI site found at 3415
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3409
    Illegal BglII site found at 2842
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3409
    Illegal suffix found at 2
    Illegal XbaI site found at 1806
    Illegal XbaI site found at 3199
    Illegal PstI site found at 2629
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3409
    Plasmid lacks a suffix.
    Illegal XbaI site found at 1806
    Illegal XbaI site found at 3199
    Illegal XbaI site found at 3424
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal PstI site found at 2629
    Illegal NgoMIV site found at 1895
    Illegal NgoMIV site found at 2178
    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
    Illegal SapI site found at 2118
    Illegal SapI site found at 2328


[edit]
Categories
//dna/chromosomalintegration
//plasmidbackbone/copynumber/medium
Parameters
chassis
copies10-12
functionLambda Red recombineering
mcsBioBrick
originp15A
resistanceK