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Part:BBa_J70500:Design

Designed by: Tom Knight   Group: Knight Lab   (2009-09-13)
Revision as of 17:36, 18 September 2009 by Tk (Talk | contribs) (→‎Design Notes)

pWV01 broad host range plasmid origin and rep protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Avoided a single Sau3AI site near the claI site, to eliminate Sau3AI sites in the final plasmid assembly.

  • PCR with primers:
    • WV-F: gtttcttcgaattcgcggccgcttctagagcgattttttattaaaacgtctcaaaatcgtttctg
    • WV-R: gtttcttcctgcagcggccgctactagtaatcattttgtttattgcaattgtattgctattaatcg

These primers were designed with Austin's GBW tool.

None of the annotated proteins have CGG codons. OrfB (rep) has a UGA stop codon, which will be read through to the next stop in M. florum.

There are no GATC sites.

Source

PCR of the plasmid pMG, kind gift from Prof. Jan Kok. This plasmid contains the ClaI fragment of the pWV01 plasmid.

References