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.
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.
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.
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.
Broad host range BioBrick plasmid derived from pRL1383a with low to medium copy number
RSF1010 is a broad-host-range plasmid first described in 1974 (Guerry 1974), with its entire sequence and gene organization subsequently described in 1989 (Scholz 1989). It is a naturally occurring 8.6kb broad-host-range plasmid in the E. coli incompatibility group Q. The conjugative transfer and stable replication of this plasmid are possible due to the mob genes with the associated origin of transfer (oriT) and the rep genes with the associated origin of vegetative replication (oriV), respectively. RSF1010 derived plasmids which include the oriV and associated Rep proteins are stably maintained in Pseudomonas (Bagdasarian 1981), Caulobacter (Umelo-Njaka et al. 2001), Erwinia, and Serratia (Leemans 1987). In addition, RSF1010 derived plasmids including the oriV, oriT and its associated rep and mob genes are transferred by conjugation to at least four cyanobacteria strains (Mermet-Bouvier 1993). These cyanobacteria strains include Synechocystis PCC6803 and PCC6714 and Synechococcus PCC7942 and PCC6301. A number of RSF1010 derived plasmids have been constructed due to the utility of this broad-host-range plasmid. For example, pSB2A, containing a 5.6kb RSF1010 derived region including the necessary mobilization and replication regions, can be transferred through conjugation to and stably maintained in Synechocystis PCC6803, PCC6714, and Synechococcus PCC7942 and PCC6301 (Marraccini 1993). The RSF1010 plasmid used in this study, pRL1383a was constructed for use in genomic studies of the diazotrophic, multicellular cyanobacterium Anabaena PCC 7120. This plasmid contains the mob and rep regions necessary for conjugation and autonomous replication, respectively. Additionally this vector is made resistant to Streptomycin and Spectinomycin due to the presence of the aadA gene (Wolk 2007). This plasmid can be mobilized by the E. coli derived self-transmissible plasmid RP4. Mobilization genes are not necessary for transfer of a mobilizable plasmid if the self-transmissible plasmid and the mobilizable plasmid share a common origin of transfer (Snyder and Champness 2007). The origin of transfer for RP4 is 99 base pairs and contains binding sites for transfer proteins encoded by RP4. For more information and a list of references, please visit: http://2008.igem.org/Team:Hawaii/Project/Part_A#Literature_Cited
The original pRL1383a MCS has been replaced by the BioBrick BBa_B0034 and its flanking BioBrick restriction sites.
Low copy BioBrick temperature sensitive standard vector
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.
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.
pACYC177 converted into BioBrick vector (formerly BBa_I739202)
partial ampicillin resistance marker (nonfunctional)
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.
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.
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.
This is a Low Copy BioBrick 2.0 Plasmid with the Ampicillin Resistance Gene ampR.
This is modified and converted from the BioBrick 1.0 Plasmid pSB4A3.
beta-lactamase ampicillin resistance gene
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.
pSB4C50: Low(?) copy BioBrick cloning/expression backbone carrying Cm 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
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.
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.
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.
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.
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.
pSB4A50: Low copy BioBrick cloning/expression backbone carrying Amp resistance
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.
pRS315 yeast shuttle vector with Biobrick MCS
A Shuttle vector for yeast. The vector is compatible with RFC10 standard and contains an ADH terminator behind the RFC10 cloning site. Ampicilin resistance for bakteria and URA3 gene for yeast selection.
contains an RFC10 compatible multiple cloning site, no restrictions sites for RFC10 in the Backbone. ADH Terminator after MCS.
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.
pCK01 converted into BioBrick vector (formerly BBa_I739202)
This plasmid is derived originally from pBeloBAC11 with multiple modifications to make it compatible with biobricks and more useable. Without modification, J61031 is a high-copy plasmid in normal laboratory strains of E. coli due to the colE1 origin of replication located within the biobrick part region. It also confers ampicillin resistance, kanamycin resistance, and constitutively-expressed RFP. Upon replacement of the SpeI/XbaI region, the colE1, ampR, and RFP features are removed, leaving behind the F plasmid replicon, the KanR gene, and an R6K origin of replication. At this point, the plasmid will replicate as a medium or high copy plasmid in pir+ or pir116 strains such as Ec100D::pir116. In non-pir strains (most lab strains), the plasmid will replicate at near-single copy levels. The restriction sites EcoRI, SpeI, NotI, and XbaI are all unique in this plasmid. There are several PstI sites, however. pBACr-NESd
kanamycin/neomycin resistance marker (NTP II marker)
beta-lactamase ampicillin resistance marker
finO incD/sopC region; contains 12 43 bp direct repeats that bind SopB protein; required for partitioning of F plasmid to daughter cells after replication as well as repression of sopAB operator;
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
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.
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.
pRS316 yeast shuttle vector with Biobrick MCS
A Shuttle vector for yeast. The vector is compatible with RFC10 standard and contains an ADH terminator behind the RFC10 cloning site. Ampicilin resistance for bacteria and URA3 gene for yeast selection.
Has an RFC10 compatible MCS, with ADH Terminator. No further RFC10 restrictions sites in the backbone.
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.
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.
pRS313 yeast shuttle vector with Biobrick MCS
A shuttle vector for yeast. The vector is compatible with RFC10 standard and contains an ADH terminator behind the RFC10 cloning site. Ampicilin resistance for bacteria and URA3 gene for yeast selection.
contains an ADH Terminator and restrictions sites for the RFC10 standard only in the multiple cloning site, no restrictions sites for RCF10 enzymes in the backbone.
pSB4A3-1 is a low copy plasmid with ampicillin resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream. A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region.
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.
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.
Multi-host replicative plasmid, for B. Subtilis and E. Coli
This plasmid is an multihost plasmid, replicated both in E. Coli at high copy number (>200) and low copy in B. Subtilis (about 20). This plasmid is ideal for importing parts in Subtilis: all the cloning can be done in E. Coli, as well as the amplification. Once the plasmid is extracted, you can incorporate it directly in B. Subtilis using electroporation. This plasmid is Ampicilyn resistant in E. Coli, and is tetracycline resistant in both B. Subtilis and E. Coli. The biobrick BBa_J04450 is inserted between the E X and SP site, has a negative cloning control in E. Coli
The EcoRI site has to be eliminated and the Biobricks prefixes and suffixes were recovered
The plasmid was obtained from Harald Putzer laboratory at the IBPC [http://www.ibpc.fr/UPR9073/equipe_HaraldJackie/AccueilJAP_HPGB.htm] . This is the fusion of the commercial plasmids pBC16-1 and pTZ19R via the site EcoRI. This site has been eliminated by the 2011 iGEM Paris Team by quick change mutagenesis, and the multicloning site was transformed into the biobrick standards.