pBS2E: Bacillus subtilis vector, lacA integration, MLS resistance
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the lacA locus and can be selected with erythromycin + lincomycin (MLS). It has a ampicillin resistance for cloning in E.coli.
Compatible with pBs1C and pBs4S in one strain.
This backbone is a BioBricked version of the B. subtilis vector pAX01. This BioBrick is part of the LMU 2012 igem project [http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks '''''Bacillus''B'''io'''B'''rick'''B'''ox] Reference: [http://www.ncbi.nlm.nih.gov/pubmed/11274134 H??rtl ''et al.'']
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(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.
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.
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.
AarI AD acceptor, pRS315, 8X LexAOPs Cyc1P, Adht1
This is an acceptor vector for AarI cloning with 8X Lex operators directly upstream of a Cyc1 promoter. The vector contains the AarI cloning cassette (AD sites, with Adh1 terminator). It is a pRS315 derived vector, for CEN ARS expression in cerevisiae (LEU2 marker).
LexA operator repeats were cloned into the PspOMI site at the 5' end of the Cyc1 promoter. The inner site was broken, thus the operator-promoter-AarI cassette fragment can be subcloned by PspOMI/SacI or Kpn1/SacI digestion into any other pRS series vector.
This is a derivative of part BBa_K106014, the AarI AD acceptor vector (pRS315, Cyc1P, Adh1t).
AarI AD acceptor vector (pRS315, Cyc1P, Adh1t)
Acceptor vector for AarI based cloning. Accepts any combination of parts beginning with an A site and ending with a D site. Expression of construct is driven by a Cyc1 promoter, with Adh1 terminator.
The pRS315 vector has been modified with an AarI cloning cassette inserted between the PspOMI and SacI sites. Please note, the actual orientation of this vector is reversed.
A modified pRS315 plasmid--CEN ARS yeast expression vector (LEU2 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.
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
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.
Integration vector for Bacillus subtilis derived from pSac-Cm
This plasmid was derived from pSac-Cm by insertion of biobrick compatible restriction sites (prefixes and suffixes), a terminator (BBa_B0015) after the suffixes sequences, and red fluorescent protein sequence (RFP) in between the prefix and suffix in its multiple cloning sites (MCS). New biobricks can be inserted into this vector by replacement of the RFP biobrick, and selection of the white colonies. This backbone has a multi host replication origin and replicates in E. coli. The plasmid is designed to integrate a cloned insert into the B. subtilis chromosome via double recombination between plasmid and chromosomal sacA sequences, and in order to check for integration a simple Phenol Red Sucrose test can be perform, if sacA is no longer functional, they will remain ph7, if they are still able to digest sucrose their pH will be under 5.
We found that that the integration plasmid that groningen send to the registry were unable to integrate in B. subtilis. This is due to the high number of mutations in the sacA integration region.
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.
AarI AD acceptor vector (pRS305, Gal1P, Adh1t)
This is an acceptor vector for AarI cloning, with A and D ends. Cloning into this plasmid will put your parts under the control of a Gal1 promoter (Gal1P) and Adh1 terminator (Adh1t). For more information regarding AarI acceptor vectors, and AarI cloning in general, please visit the UCSF 2008 wiki.
The pRS305 vector has been modified with an AarI cloning cassette inserted between the PspOMI and SacI sites. Please note, the actual orientation of this vector is reversed.
A modified pRS305 plasmid--Integrating yeast expression vector (LEU2 marker).
Transfection vector (similar to pSB1AK8)
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.
pCK01 converted into BioBrick vector (formerly BBa_I739202)
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.
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.
pACYC177 converted into BioBrick vector (formerly BBa_I739202)
partial ampicillin resistance marker (nonfunctional)
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.
Eukaryotic terminator vector (very similar to pSB1AK8)
beta-lactamase (ampicillin resistance marker)
BioBrick prefix partial (missing G at 3' end)
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.
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.
AarI AD acceptor vector (pRS315, Adh1P, Adh1t)
This is an acceptor vector for AarI cloning, with A and D ends. Cloning into this plasmid will put your parts under the control of a Adh1 promoter (Adh1P) and Adh1 terminator (Adh1t). For more information regarding AarI acceptor vectors, and AarI cloning in general, please visit the UCSF 2008 wiki.
The pRS315 vector has been modified with an AarI cloning cassette inserted between the PspOMI and SacI sites.
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.
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.
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
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
AarI AD acceptor vector (pRS315, 8X LexAOps Fig1P, Adh1t)
Acceptor for AarI AD parts, with 8X LexA operators directly upstream of the medium strong Cyc1 promoter.
LexA Ops were cloned non-directionally into the PspOMI site. Inner site is broken, thus the cassette can be subcloned from this plasmid by PspOMI/SacI digestion (for example, to change markers).
Modification of the AarI AD acceptor vector (pRS315, Cyc1P, Adh1t).
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.
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.
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.
pSBBs4S: Empty backbone for integration into Bacillus subtilis thrC locus
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the thrC locus and can be selected with Spectinomycin cassette. It has a ampicillin resistance for cloning in E.coli. It originates from the Bacillus vector pDG1731.
This plasmid can be used as empty backbone vectors like pSB1C3 but is able to integrate in B.subtilis. The clonging site also includes a rfp for selection.
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
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.
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
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.
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
Multi-host vector pTG262 converted to BioBrick vector
biobricked version of pTG262 Plasmid has a multi host replication origin and is competant in range of gram positive and gram negative bacteria.
We kindly thank Dr Mike Gasson & Dr. Claire Shearman, of the Institute of Food Research in Norwich for generously donating the pTG262 vector.
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.
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.
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)
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;
beta-lactamase ampicillin resistance marker
AarI A!D acceptor vector (pRS315, Cyc1P, Adh1t)
This is a special variant acceptor vector for AarI cloning. The "A" site used here, termed "A!" is an alternate end used when no amino-terminal fusions are tolerated. Please see the UCSF 2008 iGEM wiki for more information on AarI cloning. This was designed specifically for use with Sir3 and amino-terminal domains of Sir3.
Note that this vector will NOT accept standard AarI "A" parts.
Eukaryotic terminator vector (formerly BBa_J52017)
This vector is made of BioBrick vector pSB1AK3, which has eucaryotic terminator added.
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.
pSBBs2E: Empty backbone for integration into Bacillus subtilis lacA locus
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the lacA locus and can be selected with erythromycin + lincomycin (MLS). It has a ampicillin resistance for cloning in E.coli. 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. The clonging site also includes a rfp for selection in E.coli.
This part is an igem compatible version of the Bacillus subtilis vector pAX01
AarI AD acceptor vector (pRS315, Cyc1P, Adh1t-8XLexA Ops)
This is a variant of the AarI AD acceptor vector (pRS315, Cyc1P, Adh1t), with the addition of 8X LexA operator sites directly downstream of the Adht1 terminator.
Variant of the AarI AD acceptor vector (pRS315, Cyc1P, Adh1t)
pBR322 converted to BioBrick vector (formally BBa_I739201)
partial tetracycline resistance CDS (nonfunctional)
pBs4S: Bacillus subtilis vector, thrC integration, spec resistance
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the thrC locus and can be selected with Spectinomycin cassette. It has a ampicillin resistance for cloning in E.coli. This backbone is a BioBricked version of the B. subtilis vector pDG1731.
This vector is compatible with pBS1C and pBS2E in one strain.
This BioBrick is part of the LMU 2012 igem project BacillusBioBrickBox For handling of B. subtilis vectors, please see here. The transformation into B. subtilis is explained here.
pSB4A50: Low copy BioBrick cloning/expression backbone carrying Amp resistance
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.
The plasmid is designed to integrate a cloned insert into the B. subtilis 168 chromosome at the sacA locus. The user inserts the fragment inside the MCS whi
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
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.