nonfunctional pSC101 origin of replication
pSC101 origin of replication which is supposed to have low copy number. This was the origin used in Michael Elowitz's repressilator and in Bujard's pZS4Int-1 vector with mutations introduced to eliminate a SpeI site. It is also the origin of replication in pSB4* series plasmids.
This part is the replication origin from plasmid pBR322 which is supposed to have low to medium copy number.
pSC101ts temperature sensitive low copy origin of replication
pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30???C but stop replicating at 42???C.
The replication origin boundaries were chosen quite liberally (i.e. they might include more sequence than is strictly necessary) because a different version of this origin (BBa_I50040) was shown to be non-functional.
This part was amplified from pSB4A3-P1010.
G to A difference with canonical pSC101 vector sequence
G to C difference with canonical pSC101 vector sequence
T to C difference with canonical pSC101 vector sequence
C to T difference with canonical pSC101 vector sequence
lac promoter/rbs/rep/oriV/lac promoter/rbs/aadA/oriT
This is an intermediate construction step for the construction of the broad-host-range plasmid derived from RSF1010. The coding region was inserted into pSB1A3.
I am having trouble inserting this part into the Base Vector so the construct will be tested while still in pSB1A3.
This part was derived from RSF1010 and RP4.
Minimal pUC19-derived high copy replication origin
High copy origin of replication derived from pSB1A3. Designed to omit antibiotic resistance markers and primer binding sites. Lacks mutations that render BBa_I50020 nonfunctional as a replication origin.
We had to leave several restriction sites in because point mutations that eliminated these sites were found to render the origin nonfunctional.
R6K minimal origin of replication. Only contains gamma origin and requires pir+ strain for replication. Copy number is dependent on amount of pir protein in the cell.
pSC101 origin of replication which is supposed to have low copy number. This was the origin used in Michael Elowitz's repressilator and in Bujard's pZS4Int-1 vector with mutations introduced to eliminate a SpeI site. It is also the origin of replication in pSB4* series plasmids. This part is the reverse copy of BBa_I50040.
difference from canonical pSC101 sequence in Genbank
F plasmid backbone with BioBrick sites removed, reverse
This is a (near) single copy origin derived from the F plasmid. The naturally occurring BioBricks sites were removed with an intent to preserve the amino acid and codon frequency where possible. Similar to the pSMART VC sequence from Lucigen http://www.lucigen.com/products/Cloning_Systems/CopyRight.html. It is intended to be combined with other vector components (antibiotic resistance marker, multiple cloning site) in order to form a BioBricks vector. Relevant information can be found at http://openwetware.org/wiki/BioBrick_Parts_for_Plasmid_Engineering. This part is a reverse copy of BBa_I50000.
sequence derived from F plasmid in GenBank Accession Number AP001918
C to G mutation (eliminates XbaI site, noncoding region)
ori2 (oriS) single copy replication origin
pSC101 heat sensitive origin of replication with deletion in par locus
A derivative of pSC101 origin of replication, this heat sensitive origin has the par locus deleted, therefore allowing a rapid loss of plasmid even under the permissive temperature.
The par locus was deleted through excision of BBa_K524000 by SspI and StuI, purification of desired fragments and then
BBa_K864050 is a pSC101 origin of replication and is found in the pSB4*15 series of BioBrick vectors. This ori is an improvement of the ori found in all pSB4*5 plasmids, BBa_I50042. BBa_I50042 has been shown to have a higher copy number than expected.
This ori was created by cloning pSC101ts from pSIM5 and making a V56A mutation in repA.
Origin for Asaia and resistance to Kanamycin
Plasmids containing oriV and the corresponding Rep proteins (from RSF1010) are autonomously replicated in a broad range of hosts. It has been reported that RSF1010 derived plasmids containing oriV and the Rep proteins are stably maintained in Pseudomonas (Bagdasarian 1981), Caulobacter (Umelo-Njaka et al. 2001), Erwinia, Serratia (Leemans 1987), and several cyanobacteria strains (Mermet-Bouvier 1993) including Synechocystis PCC6803 and PCC6714 and Synechococcus PCC7942 and PCC6301.
This part is to be constructed with the Rep proteins derived from an RSF1010 plasmid.
This part is taken from the RSF1010 derived plasmid pRL1383a. A description of this plasmid can be found at the NCBI database at accession number AF403426.
pSC101 temperature sensitive origin of replication
BBa_I50052 is a pSC101 temperature sensitive origin of replication. BBa_I50052 is the temperature sensitive replication origin found in the pSB8* series of BioBrick vectors as well as the pSIM5 recombineering vector.
See design notes for Part:BBa_I50042.
This part is derived from BBa_I50042. It contains a point mutation that changes Alanine to Valine in RepA protein.
This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page] <!-- Add more about the biology of this part here
R6K origin of replication requires a pir+ or pir116 strain for replication in the absence of a second origin. In most E. coli strains, this is a silent feature. {pSB1A2-Bca1011}
PCR ca1011F/R on pG80ko (445 bp, EcoRI/SpeI) Sub into pSB1A2-I13522 (EcoRI/SpeI) Product is Bca1011 ---- ca1011F Forward Biobricking of R6K GGACTgaattcgcggccgcttctagagtgattcgcacgggcccatg ca1011R Reverse biobricking of R6K ccgctactagtaGCAGTTCAACCTGTTGATAG
p15A replication origin belongs to a different compatibility group from the common pMB1 (pBR or pUC) origin, and hence allows for co-existence with other pUC origin plasmids. When using this part, ligate it along with a proper antibiotic resistance gene and your gene of interest together to make a circular plasmid. Please do NOT include any other replication origin in your design.
Unlike pMB1 origin, which allows for chloramphenicol amplification, p15A replication origin is insensitive to chloramphenicol and remains in low copy number. This replication origin is different from the original one in that it contains several mutations, which increases the copy number to be no lower than pBR origin. In this way, both the compatibility and the convenience for use are accommodated.
p15A origin of replication which is supposed to have low to medium copy number. It is also the origin of replication in pSB3* series plasmids.
The boundaries of this part are quite liberal to ensure that this part is a functional origin.
This part was constructed by PCR amplification of pSB3K3-P1010.
F plasmid backbone with BioBrick sites removed.
This is a (near) single copy origin derived from the F plasmid. The naturally occurring BioBricks sites were removed with an intent to preserve the amino acid and codon frequency where possible. Similar to the pSMART VC sequence from Lucigen http://www.lucigen.com/products/Cloning_Systems/CopyRight.html. It is intended to be combined with other vector components (antibiotic resistance marker, multiple cloning site) in order to form a BioBricks vector. Relevant information can be found at http://openwetware.org/wiki/BioBrick_Parts_for_Plasmid_Engineering.