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.
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.
pSB1AC3 is a high copy number plasmid carrying ampicillin and chloramphenicol resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). For the version of this plasmid with a CcdB insert, see BBa_P1010.
Silent g->a mutation found during sequencing
pSB2K3-derived BioBrick plasmid free of many restriction sites
Derived from pSB2K3 with many restriction enzyme sites removed by site-directed mutagenesis (~2 dozen mutations). Plasmid is useful when there is a need to cut with a variety of different restriction enzymes. Most symmetric cutters have been removed, and some non-symmetric cutters have also including AarI, AcuI, BbsI, BbvCI, BciVI, BsaI, BsmBI, EarI, N.BstNBI, and SgrAI. Works in exactly same way as pSB2K3. See <bb_part>pSB2K3</bb_part> for more information. Note that the VF2 site has a single base mutation in the middle of it. However, VF2 has been used successfully as a primer for PCR with this plasmid.
pACYC177 converted into BioBrick vector (formerly BBa_I739202)
partial ampicillin resistance marker (nonfunctional)
pSB1A3-1 is a high copy number plasmid carrying ampicillin resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). pSB1A3 has terminators bracketing its MCS which are designed to prevent transcription from *inside* the MCS from reading out into the vector. The efficiency of these terminators is known to be < 100%. Ideally we would construct a future set of terminators for bracketing a MCS that were 100% efficient in terminating both into and out of the MCS region. A fully annotated Genbank file is attached below.
This is a high copy plasmid for use in assembly of BioBrick® standard biological parts.
The ampicillin resistance marker needed to be eliminated.
Austin Che constructed this plamid from pSB1AT3-P1010.
Verifiction forward (VF2) primer binding site
Verification reverse (VR) primer binding site
pSBBs1C: Empty backbone for integration into Bacillus subtilis amyE locus
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the amyE locus and can be selected with Chloramphenicol (cat gene). It has a ampicillin resistance for cloning in E.coli. It originates from the Bacillus vector pDG1662. This BioBrick is part of the LMU 2012 Bacillus BioBrick Box project: http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks
We removed any forbidden restriction sites by site-directed mutagenesis PCR.
This backbone is an igem compatible version of the Bacillus vector pDG1662
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
Low copy BioBrick standard vector with LacI
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.
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.
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.
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.
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.
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.
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
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.
Low to medium copy BioBrick standard vector
Erik Gullberg, Antonio Ascue Avalos, Erik Lundin
BBa_K592200 is a BioBrick standard vector with low to medium copy p15A replication origin (BBa_I50032) and spectinomycin antibiotic resistance marker, usable for Lambda Red recombineering in E coli. The E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
To make the plasmid usable for Lambda Red recombineering, all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
p15A low to medium copy replication origin BBa_I50032
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.
pBR322 converted to BioBrick vector (formally BBa_I739201)
partial tetracycline resistance CDS (nonfunctional)
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.
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.
This part was derived from pSB1AC3-P1010.
Verification forward (VF2) primer binding site
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.
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_P1003 antibiotic resistance marker.
pSB1C3F-Vector pSB1C3 with RFP-cassette in Freiburg Standard
To allow the possibility to save some nucleotides of primer-overhangs, you can use this vector. The two AgeI-restriction sites from the original RFP-cassette have been removed. The cassette is flanked by the modified Freiburg standard (http://2012.igem.org/wiki/images/b/b9/LMU-Munich_2012_Our_Freiburg_standard_FusionPrefix.pdf) that includes an optimal ribosome binding site between XbaI and NgoMIV restriction sites.
Two AgeI-restriction sites have been removed by two subsequent quikchange site-directed mutagnesis reactions. The RFP-cassette was PCR-amplified with the overhangs: prefix: GAATTCCGCGGCCGCTTCTAGATAAGGAGGAACTACTATGGCCGGC suffix: ACCGGTTAATACTAGTAGCGGCCGCTGCAGT
pSB1C3; the RFP-part is originally from Discosoma sp.
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 plasmid is based on pProTetE333 (clontech). A SpeI site was removed via an A-T mutation at position 1166 on the original plasmid, resulting in pProTetE333b. The G00100-G00101 fragment of pSB4A3-1 which includes P1010 between a biobricks multiple cloning site which itself is flanked by + strand transcriptional terminators was then cloned between the XhoI and AvrII sites of pProTetE333b. The resulting plasmid carries a ColE1 origin of replication, a Chloramphenicol resistance marker, the biobricks MCS and the ccdAB death cassette.
pProTetE333 (clontech) and biobricks parts
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.
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.
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.
This plasmid was derived from pSB1AK3-P1010.
pSB1A30: High copy BioBrick cloning/expression backbone carrying Amp resitance
pSB2K3 is a variable copy number plasmid carrying kanamycin resistance. The plasmid contains the F' replication origin and also the P1 lytic replication origin. Genes encoding proteins that are required to activate the P1 lytic replication origin are carried on the plasmid under the control of a lacI-regulated promoter. When grown in cells producing LacI the plasmid is maintained at very low copy (<10 per cell). When induced with IPTG the P1 lytic replication proteins are produced and the P1 lytic replication origin takes over, raising the copy number to very high levels (>100++ per cell). The copy control features of pSB2K3 are useful for cloning "difficult" parts while still being able to prepare large amounts of DNA. Additionally, pSB2K3 has terminators bracketing its MCS which are designed to prevent transcription from *inside* the MCS from reading out into the vector. The efficiency of these terminators is known to be < 100%. Ideally we would construct a future set of terminators for bracketing a MCS that were 100% efficient in terminating both into and out of the MCS region.
*** pSCANS via BNL *** The pSB2 series plasmids are based on the pSCANS-5 plasmid from John Dunn's group at Brookhaven National Laboratory. The pSCANS plasmids are introduced here.
MIT SBWG modification of pSCANS-5 from BNL
pBS1C: Bacillus subtilis vector, amyE integration, CAM-resistance
This vector integrates into the amyE-site of Bacillus subtilis via double recombination. The two recombination sites flank a standard BioBrick multiple cloning site (with RFP-cassette), as well as a chloramphenicol resistance cassette (5 ??g/ml). The backbone does not contain the restriction sites EcoRI, XbaI, SpeI, PstI, NgoMIV nor AgeI. The vector is replicative in Escherichia coli and provides resistance to ampicillin (100 ??g/ml). Before transformation into B. subtilis, this vector should be linearized with ScaI.
This is one of three empty vectors with compatible integration loci and resistance cassettes to allow combination of up to three constructs of choice in one single strain.
pSB4A50: Low copy BioBrick cloning/expression backbone carrying Amp resistance
pCK01 converted into BioBrick vector (formerly BBa_I739202)
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.
Transcriptionally insulated high copy BioBrick plasmid
pSB1A4 was constructed by adding double terminators ([BBa_B0015]) before the BioBrick prefix and after the BioBrick suffix in pSB1A3. This insulator blocks read-through transcription into BioBrick parts/ devices.
The double terminators were assembled from small smaller overlapping ssDNA oligos. The outer-most EcoRI and PstI sites were mutated while the inner-most BioBrick cut sites were restored.
Based upon pSB1A3; Includes two copies of [BBa_B0015] assembled from DNA oligos
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.
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.
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.
BBa_I51001 vector scaffold, BBa_I50040 replication origin and BBa_P1005 resistance marker.
pSB1AK3 is a high copy number plasmid carrying ampicillin and kanamycin resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell).
pSB3K3-1 is a medium copy plasmid with kanamycin resistance. It has a p15A pMR101-derived replication origin, copy number 20-30. (Lutz and Bujard, 1997) pSB3K3-1 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into the vectorl. 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.
pSB1AT3 is a high copy number plasmid carrying ampicillin and tetracycline resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell).