pSB4A15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ??? antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
The resistance cassette is terminated by a Lux bidirectional terminator. Adjacent to this terminator there is a designed primer binding site to where the forward primer for lambda red can be designed to bind to no matter what the insert in the cloning site is or what the resistance is. The resistance cassette is flanked by SalI and SacI restriction sites for easy switching of resistance in the backbone. The pSC101 origin of replication is flaked by NheI and MluI restriction sites for easy switching of origin of replication.
pSB4K15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and kanamycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
pSB4K15(FRT) is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and kanamycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
BioBrick base vector, as originally designed
Low copy BioBrick expression backbone carrying Kan resistance
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha
Low copy BioBrick temperature sensitive standard vector
pSB8C15 is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and Chloramphenicol antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
The resistance cassette is terminated by a Lux bidirectional terminator. Adjacent to this terminator there is a designed primer binding site to where the forward primer for lambda red can be designed to bind to no matter what the insert in the cloning site is or what the resistance is. The resistance cassette is flanked by SalI and SacI restriction sites for easy switching of resistance in the backbone. The pSC101ts origin of replication is flaked by NheI and MluI restriction sites for easy switching of origin of replication.
pSB8A15 is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
Low to medium copy BioBrick standard vector
pSB3C5 is a low to medium copy plasmid based with chloramphenicol resistance. It has a p15A-derived replication origin. pSB3C5 is derived from the BioBricks vector scaffold BBa_I51001.
BBa_I51001 BioBricks vector scaffold, p15A based origin and a version of the chloramphenicol resistance cassette that has been cleaned of restriction sites.
pSB4T3 is a low copy plasmid with tetracycline resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) It is constructed from the BioBricks vector scaffold BBa_I51001.
See http://openwetware.org/wiki/Synthetic_Biology:Vectors/pSB%2A%2A5_design for more information.
BBa_I51001 vector scaffold, BBa_I50040 replication origin and BBa_P1005 resistance marker.
pSB4C3 is a low copy plasmid based with chloramphenicol resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) pSB4C5 is derived from the BioBricks vector scaffold BBa_I51001. However, the direction of the resistance marker and replication origin is not optimal.
BBa_I51001 BioBricks vector scaffold, pSC101 based origin and a version of the chloramphenicol rsistance cassette that has been cleaned of restriction sites.
pSB3K5 is a low to medium copy plasmid based with kanamycin resistance. It has a p15A-derived replication origin. pSB3K5 is derived from the BioBricks vector scaffold BBa_I51001.
BBa_I51001 BioBricks vector scaffold, p15A based origin and a version of the kanamycin resistance cassette that has been cleaned of restriction sites.
pSB4A15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
pSB8A15 is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22
Low copy BioBrick standard vector with LacI
pSB4A15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
pSB4C15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and chloramphenicol antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
The resistance cassette is terminated by a Lux bidirectional terminator. Adjacent to this terminator there is a designed primer binding site to where the forward primer for lambda red can be designed to bind to no matter what the insert in the cloning site is or what the resistance is. The resistance cassette is flanked by SalI and SacI restriction sites for easy switching of resistance in the backbone.
pSB3T5 is a low to medium copy plasmid based with tetracycline resistance. It has a p15A-derived replication origin. pSB3T5 is derived from the BioBricks vector scaffold BBa_I51001.
BBa_I51001 BioBricks vector scaffold, p15A based origin and a version of the tetracycline resistance cassette that has been cleaned of restriction sites.
pSB4A5 is a low copy plasmid with ampicillin resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) It is derived from the BioBricks vector scaffold BBa_I51001.
See http://openwetware.org/wiki/Synthetic_Biology:Vectors/pSB%2A%2A5_design
BBa_I51001, pSC101 origin and a new version of ampR stripped of restriction sites.
pSB4A15Iq is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
pSB4S15(FRT) is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and spectinomycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
pSB8S15 is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and spectinomycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
BioBrick base vector, as revised by Codon Devices
This is a vector scaffold for constructing new BioBricks vectors. Information on the design is available at http://openwetware.org/wiki/Synthetic_Biology:Vectors/pSB%2A%2A5_design
Documented at http://openwetware.org/wiki/Synthetic_Biology:Vectors/pSB%2A%2A5_design Due to technical difficulties during synthesis, rather than including BBa_P1011 as designed, BBa_P1016 was synthesized as a component of this part. Hence, BBa_I51000 documents the original design and BBa_I51001 is the vector scaffold as synthesized.
This is a composite part generated from several basic parts obtained from a variety of sources.
pSB4A15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
pSB4K15Iq is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and kanamycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
BioBrick base vector, as synthesized by Codon Devices
BBa_I51019 is the sequence of the BioBrick base vector as synthesized by Codon Devices, Inc. in Cambridge, MA. The original design of the BioBrick base vector is available as BBa_I51000. BBa_I51001 is the sequence of the BioBrick base vector as reported by Codon Devices, Inc. although the true sequence is something like to BBa_I51019. The exact sequence of the provided DNA is difficult to determine. The corrected BioBrick base vector that was successfully constructed from BBa_I51019 is BBa_I51020.
Synthesis of the BioBrick base vector design (BBa_I51000) was problematic. We encountered two issues while trying to construct the base vector. First, troubleshooting efforts during synthesis compromised the design of our parts: failed attempts to clone the base vector into an ''E. coli'' strain intolerant of expression of the toxic protein CcdB led to an unnecessary redesign of the ''ccdB'' positive selection marker in the BioBrick base vector. As a result, a revised design for the BioBrick base vector (BBa_I51001) was specified instead. Second, faulty part design adversely impacted the synthesis process: our pUC19-based replication origin design (BBa_I50020) was similarly nonfunctional, so the revised base vector (BBa_I51001) could not be propagated as specified. We eventually determined that the provided DNA for the BioBrick base vector was actually a fusion of two slightly different copies of the base vector: one with a nonfunctional version of the pUC19 origin (BBa_I50020) and one with a functional version of the pUC19 origin (BBa_I50022). We corrected the synthesized BioBrick base vector (BBa_I51019) with molecular biology techniques to make the functional BioBrick base vector BBa_I51020 (see Methods). Commercial DNA synthesis processes currently rely on cloning, assembly and propagation of synthesized DNA in ''E. coli''. Difficulties during synthesis stemmed from the inclusion of both a ''ccdB'' positive selection marker that is toxic to most ''E. coli'' strains and a synthetic origin incapable of supporting plasmid replication of the BioBrick base vector. In general, for parts whose function are incompatible with growth and replication of ''E. coli'', the processes of DNA design and DNA synthesis cannot be easily decoupled.
pSB4A50: Low copy BioBrick cloning/expression backbone carrying Amp resistance
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena B??scher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Sch??fer, Carolin Schmelas, Silvan Schmitz, Max Waldha
pSB4S15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and spectinomycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
pSB4C50: Low(?) copy BioBrick cloning/expression backbone carrying Cm resistance
pSB8S15(FRT) is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and spectinomycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
pSB4K5 is a low copy plasmid with kanamycin resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) It is derived from the BioBricks vector scaffold BBa_I51001.
BBa_I51001 vector scaffold, BBa_I50040 replication origin and BBa_P1003 antibiotic resistance marker.
pSB4A15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. The resistance cassette is flanked by FRT sites (BBa_J61020) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase. The pSB8x15 series are especially suitable for carrying the FLP recombinase, since they can be removed by incubation at 42??C.
pSB4C15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and chloramphenicol antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
pSB8K15 is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
BioBrick base vector (for constructing pSB**5 vectors)
Documented at http://openwetware.org/wiki/Synthetic_Biology:Vectors/pSB%2A%2A5_design Due to technical difficulties during synthesis, rather than including BBa_P1011 as designed, BBa_P1016 was synthesized as a component of this part. Hence, BBa_I51000 documents the original design and BBa_I51001 is the vector scaffold as synthesized. BBa_I52002 is the base vector as corrected from the synthesized construct.