Part:BBa_K125010
Broad-Host-Range RSF1010 derived plasmid
This Broad-Host-Range plasmid is derived from RSF1010. The replication proteins, the origin of vegetative replication, and the aadA gene were inserted into the Base Vector. The MPF system has been removed and replaced with the RP4 origin of transfer.
This plasmid has been converted to BioBrick format and is also more compact that the original version. The replication proteins with the origin of vegetative replication allows for autonomous replication in a broad range of gram negative bacteria. The oriT allows for mobilization by the self-transmissible plasmid RP4. The aadA region allows resistance to Spectinomycin and Streptomycin. The Base Vector provides an insulated BioBrick cloning site containing a high copy origin and the ccdB death gene which are both removed when a part in cloned in. This allows one to make a lot of the plasmid for cloning, and a means for positive selection.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 5885
Illegal suffix found in sequence at 4559 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 5885
Illegal NheI site found at 4735
Illegal NheI site found at 5684
Illegal SpeI site found at 4560
Illegal PstI site found at 4574
Illegal NotI site found at 4567
Illegal NotI site found at 5891 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 5885
Illegal BglII site found at 6341 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 5885
Illegal suffix found in sequence at 4560 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 5885
Illegal XbaI site found at 5900
Illegal SpeI site found at 4560
Illegal PstI site found at 4574
Illegal NgoMIV site found at 4299 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 6229
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