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.
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.
pBR322 converted to BioBrick vector (formally BBa_I739201)
partial tetracycline resistance CDS (nonfunctional)
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
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
with a Ptet promoter controled by TetR, a MCS, T1 terminator, p15A ori and Cm resistence gene. copy number: 15~20
to measure the efficiency of different SD signal, the plasmid has a MCS downstream of Ptet where can insert reporters along with SD signal. There is a T1 terminator downstream of MCS to terminate transcription starting from Ptet. Ptet is repressed by TetR carried on this plasmid and can be turned on by adding aTc.
pBR322 has Pst I site in its Amp resistant gene, so you can't introduce Biobrick site(Pst I included) in this plasmid. For that reason, we introduced a silent mutation at the site. The mutation is A546C(DNA sequence) in the Amp resistant gene. Introducing this mutation allowed us to aim at constructing pBR standard plasmid.
A546C(DNA sequence) = A182A(amino acid sequence)
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.
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.
Low to medium copy Lambda Red recombineering compatible plasmid
BBa_K592202 is a BioBrick friendly vector with low to medium copy p15A replication origin (BBa_I50032) and a kanamycin 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, and the kanamycin resistance marker is flanked by FRT sites so that the resistance marker can be removed after chromosomal integration using the FLP recombinase.
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. To be able to remove the antibiotic resistance marker after chromosomal integration, the kanamycin marker from the pKD4 plasmid (Datsenko and Wanner, 2000) was used. The resistance marker can be removed after chromosomal integration using the FLP recombinase, leaving only the plasmid insert and a scar behind.
The P22 late terminator come from the genome of the Salmonella phage P22 (GenBank: AF217253.1, 19662..19702), and the kanamycin resistance marker flanked by FRT sites comes from the pKD4 plasmid (Datsenko and Wanner, 2000).