Part:BBa_K864019

Low copy BioBrick temperature sensitive standard vector

pSB8S15 is a BioBrick standard vector with low copy pSC101ts replication origin BBa_K864050 (~5 copies per cell) and spectinomycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30’C but stop replicating at 42’C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.

The pSB8X15 series are especially suitable for carrying the FLP recombinase, since they can be removed by incubation at 42°C.

Sequence and Features

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 3772
  Illegal NheI site found at 2425
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 3778
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3772
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 3772
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 3772
  Plasmid lacks a suffix.
  Illegal XbaI site found at 3787
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 2319
  Illegal NgoMIV site found at 2637
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.