Difference between revisions of "Part:pSB3K3:Design"
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<partinfo>pSB3K3 short</partinfo>
 
<partinfo>pSB3K3 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The sequence immediately downstream of the p15a origin in this plasmid is an almost exact match to the AmpR cassette (see BBa_P1002).  It seems likely that the orignal definition of the p15a origin was generous and could likely be truncated.
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===Subversions===
  
 +
K2491030 (https://parts.igem.org/Part:BBa_K2491030)
  
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In 2017, Philippe Hansen-Estruch from the CCA_San_Diego team created a new plasmid based off the pSB3K3 plasmid. The p15A origin of replication was replaced with a RK2 origin of replication. The goal of this improvement was to replace p15a origin of replication (ori) with the RK2 ori to generate a shuttle vector as a broad host range vector (i.e. can replicate in E.coli and other microorganisms).
  
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The RK2 origin of replication is a broad-host-range plasmid belonging to the incP incompatibility group that can be maintained in a large number of bacteria.  The minimal region for replication and maintenance consists of an origin of replication, oriV, and the plasmid replication initiator protein TrfA protein, that activates oriV. The copy number of RK2 is about 4-7 per cell in E. coli, 3 in Pseudomonas aeruginosa, and 4-7 in Agrobacterium.
  
 
===Source===
 
===Source===

Latest revision as of 03:40, 24 October 2017

Low to medium copy BioBrick standard vector


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 2729
    Illegal NheI site found at 1384
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2735
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2729
    Illegal XhoI site found at 177
    Illegal XhoI site found at 1020
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 2729
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 2729
    Plasmid lacks a suffix.
    Illegal XbaI site found at 2744
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal AgeI site found at 1470
    Illegal AgeI site found at 1793
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2498


Design Notes

The sequence immediately downstream of the p15a origin in this plasmid is an almost exact match to the AmpR cassette (see BBa_P1002). It seems likely that the orignal definition of the p15a origin was generous and could likely be truncated.


Subversions

K2491030 (https://parts.igem.org/Part:BBa_K2491030)

In 2017, Philippe Hansen-Estruch from the CCA_San_Diego team created a new plasmid based off the pSB3K3 plasmid. The p15A origin of replication was replaced with a RK2 origin of replication. The goal of this improvement was to replace p15a origin of replication (ori) with the RK2 ori to generate a shuttle vector as a broad host range vector (i.e. can replicate in E.coli and other microorganisms).

The RK2 origin of replication is a broad-host-range plasmid belonging to the incP incompatibility group that can be maintained in a large number of bacteria. The minimal region for replication and maintenance consists of an origin of replication, oriV, and the plasmid replication initiator protein TrfA protein, that activates oriV. The copy number of RK2 is about 4-7 per cell in E. coli, 3 in Pseudomonas aeruginosa, and 4-7 in Agrobacterium.

Source

References