Plasmid_Backbone
pSac-Cm_BB

Part:BBa_K818000

Designed by: Emeraldo Jo   Group: iGEM12_Groningen   (2012-08-22)

Integration vector for Bacillus subtilis derived from pSac-Cm

This plasmid was derived from pSac-Cm by insertion of biobrick compatible restriction sites (prefixes and suffixes), a terminator (BBa_B0015) after the suffixes sequences, and red fluorescent protein sequence (RFP) in between the prefix and suffix in its multiple cloning sites (MCS). New biobricks can be inserted into this vector by replacement of the RFP biobrick, and selection of the white colonies. This backbone has a multi host replication origin and replicates in E. coli. The plasmid is designed to integrate a cloned insert into the B. subtilis chromosome via double recombination between plasmid and chromosomal sacA sequences. For more info about this biobrick and its usage in B. subtilis, please visit: Groningen 2012 BioBrick page

Usage and Biology

This plasmid works well in E. coli and B. subtilis. This plasmid can be used as cloning tool in E. coli by selecting the white colonies from the red colonies after transformation (the plasmid in the red colonies has no insert; the RFP sequence is not replaced by the insert).
















red-white colonies

Sequence and Features

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 EcoRI site found at 5207
    Illegal XbaI site found at 5218
    Illegal SpeI site found at 1
    Illegal PstI site found at 12


[edit]
Categories
//chassis/multihost
//chassis/prokaryote/bsubtilis
//plasmidbackbone/operation
Parameters
chassisMulti host
mcsBiobrick - BBa_J04450
originMulti host
resistanceAC