Plasmid_Backbone

Part:BBa_K535007:Design

Designed by: iGEM11_UNAM-Genomics_ Mexico   Group: iGEM11_UNAM-Genomics_Mexico   (2011-09-25)
Revision as of 02:57, 26 September 2011 by Alecanda (Talk | contribs) (New page: __NOTOC__ <partinfo>BBa_K535007 short</partinfo> <partinfo>BBa_K535007 SequenceAndFeatures</partinfo> ===Design Notes=== We amplified the RFP Coding Device [https://parts.igem.org/Pa...)

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pBBR1MCS-5 -> broad-host-range cloning vector


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Illegal EcoRI site found at 5006
    Illegal XbaI site found at 5021
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5006
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3127
    Illegal NotI site found at 5012
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5006
    Illegal BglII site found at 3927
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Illegal suffix found at 2
    Illegal EcoRI site found at 5006
    Illegal XbaI site found at 5021
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5006
    Illegal XbaI site found at 5021
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 855
    Illegal AgeI site found at 695
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 3449


Design Notes

We amplified the RFP Coding Device BBa_J04450 including the prefix and the suffix from the pSB1T3 plasmid and we added apaI and sacI restriction sites at the ends of the sequence.

Then we made a ligation by sticky ends with pBBR1MCS-5 digested by apaI and sacI, with that we eliminate as many as possible restriction sites from the MCS region.

We use the sequence as reported by Kovach et al. (1995).

Source

This is one of the antibiotic-resistant derivatives of the broad-host-range cloning vector pBBR1MCS.

References

  • Michael E. Kovach, Philip H. Elzer, D. Steven Hill, Gregory T. Robertson, Michael A. Farris,R. Martin Roop, Kenneth M. Peterson (1995) Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 166, 175-176.