Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Brice Enjalbert
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-15 03:11:34
OriKan: a composite part to turn any pSB1C3 into a Bacillus plasmid
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1937002_sequence (Version 1) |
Description
Part: BBa_K1937002 (pSB1C3-OriKan)(Chassis E. coli/B. subtilis, carrier plasmid pSB1C3)
Length: 2510 bp
While Bacillus subtilis is of huge interest for a growing number of iGEM projects, it is not easy to develop new parts as it is required that they registered after sub-cloning in the E. coli plasmid pSB1C3. During the iGEM Toulouse 2016 project, we had the idea to create a part that could turn any pSB1C3-based plasmid in a shuttle vector (E. coli/B. subtilis).
This BioBrick is a part developed by the Toulouse 2016 iGEM team (http://2016.igem.org/Team:Toulouse_France)
Notes
We successfully transformed E. coli and selected clones on chloramphenicol. Likewise, we successfully transformed B. subtilis and selected clones on kanamycine. Presence of the pSB1C3-OriKan plasmid was demonstrated in B. subtilis by PCR checking. The part sequence has been verified by sequencing.Source
The BBa_K1937002 part contains the repU origin of replication of B. subtilis and the kanamycin resistance gene for B. subtilis. It was obtained by amplifying the repU-Kan region of pSBBsOK_P (BBa_K1351040) using primers OriKan forward (5??? cacagaatcaggggataacgcaggaaagaaACATGTAGTTATAAGTGACTAAACAAATAACTAAATAGATGGG) and OriKan reverse (5??? gttcctggccttttgctggccttttgctcaACATGTCGCAAAATGGCCCGATTTAAG). The resulting fragment was sub-cloned in the pSB1C3 between the EcoRI and PstI restriction sites.| Sequence Annotation | Location | Component / Role(s) |
| Kananmycin resistance (Bacillus) RepU Bacillus origin | 1595,2365 431,1435 | CDS feature/protein sequence_feature feature/misc |