Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Grace Kwan
Date created: 2008-07-23 11:00:00
Date modified: 2015-05-08 01:09:44
Broad host range BioBrick plasmid derived from pRL1383a with low to medium copy number
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | BBa_K125000_sequence (Version 1) |
Description
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
Notes
The original pRL1383a MCS has been replaced by the BioBrick BBa_B0034 and its flanking BioBrick restriction sites.Source
Plasmid is derived from pRL1383a.| Sequence Annotation | Location | Component / Role(s) |
| P5 promoter VR VF2 oriT oriR RSF1010 derived region omega interposon spectinomycin marker T7 promoter cosN loxP T4 32 terminator T4 32 terminator BioBrick suffix BioBrick prefix | 8200,8230 78,97 8908,8927 3476,3633 2819,3243 2664,8356 463,1255 933,1943 8880,8898 8616,8705 8831,8864 343,374 2249,2280 1,21 9026,9047 | promoter feature/promoter feature/primer_binding primer_binding_site feature/primer_binding primer_binding_site sequence_feature feature/misc feature/misc sequence_feature feature/misc sequence_feature feature/cds CDS feature/cds CDS promoter feature/promoter sequence_feature feature/dna sequence_feature feature/dna stem_loop feature/stem_loop feature/stem_loop stem_loop sequence_feature feature/dna feature/dna sequence_feature |