Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Sri Kosuri
Date created: 2004-05-03 11:00:00
Date modified: 2015-05-08 01:14:55
Inducible copy number BioBrick plasmid
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | pSB2K3_sequence (Version 1) |
Description
pSB2K3 is a variable copy number plasmid carrying kanamycin resistance. The plasmid contains the F' replication origin and also the P1 lytic replication origin. Genes encoding proteins that are required to activate the P1 lytic replication origin are carried on the plasmid under the control of a lacI-regulated promoter. When grown in cells producing LacI the plasmid is maintained at very low copy (<10 per cell). When induced with IPTG the P1 lytic replication proteins are produced and the P1 lytic replication origin takes over, raising the copy number to very high levels (>100++ per cell). The copy control features of pSB2K3 are useful for cloning "difficult" parts while still being able to prepare large amounts of DNA.Additionally, pSB2K3 has terminators bracketing its MCS which are designed to prevent transcription from *inside* the MCS from reading out into the vector. The efficiency of these terminators is known to be < 100%. Ideally we would construct a future set of terminators for bracketing a MCS that were 100% efficient in terminating both into and out of the MCS region.
Notes
- pSCANS via BNL ***
The pSB2 series plasmids are based on the pSCANS-5 plasmid from John Dunn's group at Brookhaven National Laboratory. The pSCANS plasmids are introduced here.
Source
MIT SBWG modification of pSCANS-5 from BNL| Sequence Annotation | Location | Component / Role(s) |
| kanR prefix suffix F1 Replication Origin repE (rev) oriS Replication Origin (rev) lacOP repL nnpp promoters VR VF2 deletion of two bases terminator stem loop E. coli his operon terminator terminator stem loop end of terminator | 3433,4248 4404,4425 1,21 242,290 617,1372 1463,1679 2037,2211 2289,3134 3242,3393 157,176 4286,4305 34,35 30,64 22,93 4375,4394 4362,4365 | CDS feature/cds engineered_region feature/BioBrick engineered_region feature/BioBrick promoter feature/promoter feature/cds CDS promoter feature/promoter feature/operator operator feature/cds CDS feature/promoter promoter primer_binding_site feature/primer_binding primer_binding_site feature/primer_binding feature/misc sequence_feature stem_loop feature/stem_loop sequence_feature feature/misc feature/stem_loop stem_loop feature/misc sequence_feature |