Difference between revisions of "Part:BBa K864051"
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<partinfo>BBa_K864051 short</partinfo>
 
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BBa_K864051 is a pSC101ts origin of replication and is found in the pSB8*15 series of BioBrick vectors. This ori is an improvement of the ori found in all pSB4*5 plasmids, [[Part:BBa_I50042|BBa_I50042]]. [[Part:BBa_I50042|BBa_I50042]] has been shown to have a higher copy number than expected.
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BBa_K864051 is a pSC101ts origin of replication and is found in the pSB8*15 series of BioBrick vectors. This ori is identical to <partinfo>K864051</partinfo>, 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.
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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.  
  
 
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<partinfo>BBa_K864051 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K864051 SequenceAndFeatures</partinfo>
  
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===References===
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[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
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[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
  
 
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Revision as of 23:49, 28 September 2012

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