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===Stringent vs. relaxed replication=== | ===Stringent vs. relaxed replication=== | ||
Plasmid replication control is usually controlled by balancing the levels of a positive and a negative regulator of replication. For some plasmids (pMB1/colE1 replicons) the positive regulator is an RNA and in others (e.g. pSC101) it is a protein. Plasmids with a protein positive regulator will not replicate in the abscence of protein production - ''stringent control'' (although not the same as the stringent response due to a shortage of loaded tRNAs). Plasmids with an RNA positive regulator will continue to replicate in the abscence of protein production. This is termed ''relaxed control''. High yields of plasmid may be obtained by halting protein production (via chloroamphenicol) when the culture reaches a high density and then continuing incubation for a number of hours. This might be of practical relevance when prepping the 1 and 3 series of Synthetic Biology plasmids.--[[User:Bcanton|BC]] 19:05, 3 Sep 2005 (EDT) | Plasmid replication control is usually controlled by balancing the levels of a positive and a negative regulator of replication. For some plasmids (pMB1/colE1 replicons) the positive regulator is an RNA and in others (e.g. pSC101) it is a protein. Plasmids with a protein positive regulator will not replicate in the abscence of protein production - ''stringent control'' (although not the same as the stringent response due to a shortage of loaded tRNAs). Plasmids with an RNA positive regulator will continue to replicate in the abscence of protein production. This is termed ''relaxed control''. High yields of plasmid may be obtained by halting protein production (via chloroamphenicol) when the culture reaches a high density and then continuing incubation for a number of hours. This might be of practical relevance when prepping the 1 and 3 series of Synthetic Biology plasmids.--[[User:Bcanton|BC]] 19:05, 3 Sep 2005 (EDT) | ||
===Online Vector Databases=== | |||
You can often find vector information at [http://www.ncbi.nlm.nih.gov/ NCBI,] either directly or in their list of vectors screened for contamination of new sequence at [http://www.ncbi.nlm.nih.gov/VecScreen/replist.html Vecscreen]. | |||
[http://seq.yeastgenome.org/vectordb/ VectorDB] contains information about many common vectors, including yeast vectors. | |||
[http://www.embl-hamburg.de/display?file=~geerlof/webPP/vectordb/vector_index.html EMBL] maintains a large database of vectors. | |||
For eukaryotic vectors (Fish, Xenopus) see [http://beckmancenter.ahc.umn.edu/cgi-bin/plasmidlookup.pl Minnesota]. | |||
The [http://www-rcf.usc.edu/~forsburg/vectors.html Forsburg Lab] maintains a list of Fisson Yeast vectors. | |||
[http://www.promega.com/vectors/ Promega] maintains a list of their vectors. | |||
[http://www.neb.com/nebecomm/products/category24.asp?#29 NEB] maintains a list of common vectors. | |||
[http://www.epibio.com/sequences.asp Epicentre] also maintains its own list. | |||
===Annotation=== | ===Annotation=== | ||
