Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Brice Enjalbert
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-18 09:20:09
pSBBsOK-mini, a small replicative plasmid for Bacillus subtilis
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1937001_sequence (Version 1) |
Description
Part: BBa_K1937001 (pSBBsOK-mini)(Chassis E. coli, carrier plasmid pSB1C3, part destined for use in Bacillus subtilis)
Length: 5857 bp
This part is an expression vector for Bacillus subtilis. It is replicative both in E.coli and B.subtilis. It has an ampicillin resistance for cloning in E.coli and kanamycin resistance for selection in B. subtilis.
This BioBrick is a part developed by the Toulouse 2016 iGEM team (http://2016.igem.org/Team:Toulouse_France)
We successfully transformed E. coli and selected clones on ampicillin. Likewise, we successfully transformed B. subtilis and selected clones on kanamycine. The plasmid sequence has been entirely verified by sequencing. The capacity to sub-clone in this new backbone has been demonstrated (part BBa_K1937004, BBa_K1937006, BBa_K1937008 and BBa_K1937010).
Notes
The pSBBsOK-mini is a reduction of the pSBBsOK -P plasmid (BBa_K1351040). Its size is only 5827 pb compared to the entire pSBBsOK -P which is 9358 pb long.Source
The part was created by amplifying a 5827pb fragment from the pSBBsOK -P plasmid (BBa_K1351040) using primers pSBBsOK Forward (5??? ctgctggctagcTACTAGTAGCGGCCGCTGCAGGCGCAACGCAATTAATGTGAGTTAGG) and pSBBsOK Reverse (5??? gacgacgctagcCTCTAGAAGCGGCCGCGAATTcAAATCAGCAAGGGACAGGTAG). The PCR product was digested by NheI and ligated on himself. The resulting plasmid is a lighter shuttle plasmid (E. coli and B. subtilis) with all the properties of the iGEM pSBA13 plasmid (prefix, suffix, verification sequences???).| Sequence Annotation | Location | Component / Role(s) |
| Bleomycin resistance (B. subtilis) Ampicillin resistance (E. coli) ColE1 origin (E. coli) Kanamycin resistance (B. subtilis) Prefix Suffixe repU (Bacillus subtilis origin) Verification forward (VF2) primer binding site Verification reverse (VR) primer binding site | 5202,5600 1564,2223 784,1466 4209,4979 257,278 285,305 3045,4049 2546,2565 685,704 | feature/protein CDS feature/protein CDS sequence_feature feature/misc feature/protein CDS sequence_feature feature/dna sequence_feature feature/dna feature/misc sequence_feature primer_binding_site feature/primer_binding primer_binding_site feature/primer_binding |