Part:BBa_J107112:Design

pSC101ts origin with SpeI removed

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 1438
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The origin has been retrieved from pAH123 vector [GenBank: AY048726]. The sequence was extended to the upstream NcoI restriction site and to the downstream stop codon of the ampicillin resistance gene in pAH123. We also modified the origin region by removing the SpeI site, according to the modification present in the available sequence of pSC101 (BBa_I50042), to ensure that the nucleotide changes do not affect the replication origin functioning, e.g., in the widely used pSB4C5 vector. The sequence of pAH123 was obtained via DNA sequencing, since its submitted DNA sequence [GenBank: AY048726] contains errors, e.g., in the temperature-sensitive mutation. The read sequence was then modified to remove SpeI site.

Source

Synthetic DNA inspired by the pAH123 vector.

References

For pAH123 description, see doi: 10.1128/JB.183.21.6384-6393.2001