Difference between revisions of "Part:pSB3K3:Design"
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K2491030 (https://parts.igem.org/Part:BBa_K2491030) | K2491030 (https://parts.igem.org/Part:BBa_K2491030) | ||
| − | In 2017, Philippe Hansen-Estruch from the CCA_San_Diego 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). | + | 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. | 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. | ||
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===Source=== | ===Source=== | ||
Latest revision as of 03:40, 24 October 2017
Low to medium copy BioBrick standard vector
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 2729
Illegal suffix found at 2 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.