OriT-R (Origin of transfer for the R-plasmid nic region)
OriTR, the R plasmid nic region, is where the relaxosome nicks the plasmid and conjugative transfer by R plasmid machinery begins.
OriT-RP4 (Origin of transfer for the RP4 plasmid)
Relaxation region of self-transmissible plasmid RP4. When this part is added to a plasmid, it is transferred by RP4 through conjugation.
This part is similar to J01003, which has no experience and is much longer. In the process of designing and testing this part, it will be compared to J01003.
F plasmid with OriT and TraJ knocked out by lambda-red
The relaxase / helicase TraI of the IncP beta plasmid R751. Essential of conjugative transfer. This part nicks oriT. It should be placed on a message plasmid along with oriT, and will enable transfer of the message plasmid in the presence of a modified R751 helper plasmid such as BBa_I714037.
This part comes from http://www.ncbi.nlm.nih.gov/nuccore/70907607?from=48797&to=51040&report=gbwithparts and corresponds to range 48797..51040 of the R751 sequence.
TrbA transcription factor for the Tra2 operon in incP alpha conjugation
Detailed description to be entered by Scott
trbK (entry exclusion) of IncP beta R751 plasmid
This part encodes for trbK, a small 75 amino acid cytoplasmic membrane lipoprotein which handles the entry exclusion in the R751 plasmid. Expression of this part in a cell without R751 will significantly reduce the transfer frequency from R751 donors. This is designed to prevent redundant transfers from occurring. Note that the presence of entry exclusion has been linked to the prevention of lethal zygosis. If the entry exclusion (Eex) index is defined as the transfer frequency for receiver cells not containing the trbK protein divided by the transfer frequency for receiver cells containing the trbK protein than for R751 Eex index = 44,000 (See Table 5 of Haase, J., Kalkum, M. & Lanka, E. TrbK, a small cytoplasmic membrane lipoprotein, functions in entry exclusion of the IncP alpha plasmid RP4. J. Bacteriol. 178, 6720???6729 (1996).[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=178567])
This sequence can be found at http://www.ncbi.nlm.nih.gov/protein/10955220
OriT-R (Origin of Transfer for the R751-plasmid nic region)
OriT-R751 (Origin of Transfer for the R751-type conjugative plasmid) is the nic region, where the relaxosome nicks the plasmid and conjugative transfer by R751 plasmid machinery begins via rolling circle replication.
Enterobacter aerogenes plasmid R751 NCBI:NC_001735
TraJr protein (controls conjugative transfer in RP4 plasmid)
TraJR controls conjugative transfer in R plasmid
Intermediate of the oriTr (RP4 origin of transfer) and R6K (conditional origin of replication) pSB1A2-Bca9009
R plasmid with OriT and TraJ knocked out by lambda-red
Vector containing Hisx6 and Sortase tag LPETGG
This vector is constructed to simplify the usage of the transpeptidase Sortase A for future iGEM teams. Following a BamHI restriction site we constructed a synthetic sequence consisting of three main elements: 1) A linker (GSSS) to allow for limited mobility of the attached peptides and facilitate correct folding 2) A Sortase A tag (LPETGG) that is used to conjugate the proteins to poly-glycines 3) A standard Hisx6 tag commonly used for Immobilized Metal Affinity Chromatography (IMAC) This sequence follows a BamHI restriction site which can be used by teams to clone in whichever peptide they would like to conjugate using Sortase A.
BamHI is a very convenient restriction enzyme for our application as its restriction site will code for a glycine and a serine, both of which are often used in linkers and tend to be unproblematic in such sequences.
The plasmid was constructed using the iGEM-supplied pSB1C3 and a synthetically produced sequence.
OriT-F (Origin of Transfer for the F-plasmid nic region)
OriTf (Origin of Transfer for the F-type conjugative plasmid) is the nic region, is where the relaxosome nicks the plasmid and conjugative transfer by F plasmid machinery begins via rolling circle replication. This particular plasmid has been derived from a F-type plasmid donated by Professor Laura Frost University of Alberta. NOTE: OriTf is NOT the same as the host cell's native Origin of Transfer (this is a separate site within the bacterial host's main genome).
OriT-S (Origin of Transfer for the pSC101-plasmid nic region)
OriT-S (Origin of Transfer for the pSC101-type conjugative plasmid) is the nic region, where the relaxosome nicks the plasmid and conjugative transfer by pSC101 plasmid machinery begins via rolling circle replication.
This part was constructed to compare the oriTr, which is currently available in the registry to oriT which was recently designed to perform the same function. The oriTr was added to LacZ part so that conjugation could be monitored by blue/white selection.
This part will be tested by blue/white screening.
This part was built from oriTr and LacZ.
PhoA: signal sequence for peri plasmic expression
Marie Burghard, Jascha Diemer, Adrian Eilingsfeld, Philipp Rottmann, Rene Sahm, Arne Wehling
PhoA is an N-terminal signal sequence for expression into the peri plasma. It derives from the native phosphatase A of E.coli and is similar to pelB. We used it to express our fusion protein (BBa_K808030)
As an signal peptide, our phoA has no stop codons and needs to be fused at the N-Terminus.
Our phoA BioBrick is derived from the pEST100 vector which was used in the following publication: Becker, S., Theile, S., Heppeler, N., Michalczyk, A., Wentzel, A., Wilhelm, S., Jaeger, K.-E., et al. (2005). A generic system for the Escherichia coli cell-surface display of lipolytic enzymes. FEBS letters, 579(5), 1177???82. doi:10.1016/j.febslet.2004.12.087
The oriT was synthesized and J33207 was extracted from the registry.
The oriT sequence was taken from the self-transmissible plasmid RP4. LacZ is available as a part in the registry.
TraJF controls conjugative transfer in F plasmid
OriT-F (Origin of Transfer for the F-type conjugative plasmid) is the nic region, where the relaxosome nicks the plasmid and conjugative transfer by F plasmid machinery begins via rolling circle replication.
This OriT-F is the same as the host cell's native Origin of Transfer, Please compare the sequence with J01002.
Origin-of-transfer nic region excised from Plasmid F genomic DNA.(NCBI:AP001918)