Part:BBa_K592202

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.

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