Part:BBa_K864051

pSC101ts temperature sensitive low copy origin of replication

BBa_K864051 is a pSC101ts origin of replication and is found in the pSB8*15 series of BioBrick vectors. This ori is identical to BBa_K864051, except for a mutation as described by [1]. This mutation changes one amino acid in the regulatory protein repA from alanine to valine, changing its binding properties.

Strains carrying the pSC101 plasmids can be grown stably at 30°C, but the plasmid will be lost at 42°C. For removing the plasmid, the strain can be streaked on a antibiotics-free LB agar plate and grown at 42° C overnight. Plasmid loss can be confirmed by streaking the new colonies on a plate with the relevant antibiotic. Transformants of pSB8 plasmids can, however, still recover from transformation at 37° without noticeable plasmid loss.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2130
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[http://dx.doi.org/10.1016/0022-2836(84)90352-8] K.A. Armstrong, R. Acosta, E. Ledner, Y. Machida, M. Pancotto, M. McCormick, H. Ohtsubo, E. Ohtsubo: "A 37×10(3) molecular weight plasmid-encoded protein is required for replication and copy number control in the plasmid pSC101 and its temperature-sensitive derivative pHS1" J. Mol. Biol., 175 (1984), 331€“348

[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC235445/] T Hashimoto-Gotoh, M Sekiguchi: "Mutations of temperature sensitivity in R plasmid pSC101" J. Bacteriol. 131.2 (1977) 405-412