Plasmid_Backbone
pSac-Cm_BB

Part:BBa_K818000:Design

Designed by: Emeraldo Jo   Group: iGEM12_Groningen   (2012-08-22)
Revision as of 11:45, 21 September 2012 by Emeraldo88 (Talk | contribs) (→‎References)

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Integration vector for Bacillus subtilis derived from pSac-Cm


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5207
    Illegal XbaI site found at 5218
    Illegal SpeI site found at 1
    Illegal PstI site found at 12
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5207
    Illegal SpeI site found at 1
    Illegal PstI site found at 12
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5207
    Illegal BamHI site found at 180
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5207
    Illegal XbaI site found at 5218
    Illegal SpeI site found at 1
    Illegal PstI site found at 12
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5207
    Illegal XbaI site found at 5218
    Illegal SpeI site found at 1
    Illegal PstI site found at 12
    Illegal NgoMIV site found at 4124
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 2051
    Illegal SapI.rc site found at 3133


Design Notes

This part backbone plasmid design was derived from pSac-Cm Bacillus subtilis backbone plasmid. It has a terminator in its multiple cloning site and red fluorescent protein between the prefix and suffix sequences. It will integrate to B. subtilis chromosome via double recombination from the plasmid and chromosomal sacA sequences.

Source

Molecular Genetics Group at the University of Groningen, the Netherlands.

References

Middleton, R., Hofmeister, A. New shuttle vectors for ectopic insertion of genes into Bacillus subtilis. Plasmid Volume 51, Issue 3, May 2004, Pages 238–245

Becker E, Herrera NC, Gunderson FQ, Derman AI, Dance AL, Sims J, Larsen RA, Pogliano J. DNA segregation by the bacterial actin AlfA during Bacillus subtilis growth and development. EMBO J. 2006 Dec 13;25(24):5919-31. Epub 2006 Nov 30.