Plasmid_Backbone

Part:BBa_K415049:Design

Designed by: Shawn Pan   Group: iGEM10_MIT   (2010-10-26)


Cm Backbone with pBR322 Derived Origin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2405
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2411
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2405
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 2405
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 2405
    Plasmid lacks a suffix.
    Illegal XbaI site found at 2420
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal SapI site found at 1414


Design Notes

The extra reverse sequencing site needs to be removed in the final version (pSB6C5). The resistance P1004 was placed before the origin operon K415039 to provide a promoter to the copy-number regulating protein ROP.


Source

This backbone was constructed with the standard biobrick plasmid construction method. The origin is K415039 and the chloramphenicol resistance is from P1004.

After several failures in getting the correct colonies with the ccdB containing base vector I51020, the appropriate base vector region was obtained by PCR of an existing version 5 backbone containing RFP (pSB3T5). The RFP version of the base vector allowed for easy screening for correct colonies.

References