Part:BBa_J99000:Design

pSWK oriTRP4; [CmR]; oriVR6K; lambda_attP

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 2209
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 2215
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2209
  Illegal BglII site found at 917
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 2209
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 2209
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2224
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 2178
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  

Design Notes

attP recombination site of the lambda phage was PCR amplified with oligos 5'-TGGAGCTCAAATCAAATAATGATTTTATTT-3' and 5'-GCTGGAGCTCCATGGTACGCGTGCTAGAGGCA-3', and cloned at the SacI site of pSW23T (Demarre et al., 2005). The multiple cloning site was then replaced with the BioBrick strandard restriction sites through inverse PCR with oligos 'GCTTCTAGAGTACTAGTAGCGGCCGCTGCAGGCGGTGGAGCTCAAATCAAATAATG' and 'AGTACTCTAGAAGCGGCCGCGAATTCTATCAAGCTTATCGATACCGTCGACG'

Source

This plasmid was constructed from pSW23T (Demarre et al., 2005)

References

Demarre, G. et al. A new family of mobilizable suicide plasmids based on broad host range R388 plasmid (IncW) and RP4 plasmid (IncPalpha) conjugative machineries and their cognate Escherichia coli host strains. Research in microbiology 156, 245-55(2005).

Valens, M. et al. Macrodomain organization of the Escherichia coli chromosome. The EMBO journal 23, 4330-41(2004).