Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Daniel Goodman
Date created: 2008-10-27 12:00:00
Date modified: 2015-05-08 01:08:37
Gram-positive Shuttle Vector for Episomal Expression
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_K090401_sequence (Version 1) |
Description
This episomal plasmid is capable of transformation into both Gram-negative and Gram-positive organisms. It replicates as an episomal plasmid and does not integrate into the genome.For selection in E. coli, use Ampicillin, 100 ??g per mL. For selection in B. subtilis (and other similar gram-positive organisms), use Chloramphenicol, 5 ??g per mL (not 35).
Notes
We had to remove a PstI site inside the coding region of the beta-lactamase gene coding for Ampicillin resistance, and thus the backbone had to be PCRed from two parts, with a joining primer filling in cut site with a degenerate codon.Source
This part was made via InFusion assembly from many separate pieces. The backbone comes from PAD123 (Bacillus Stock Center catalog number ECE166), and contains replication origins for B. subtilis and E. coli, as well as resistance markers for Ampicillin and Chloramphenicol.| Sequence Annotation | Location | Component / Role(s) |
| Biobrick Suffix Biobrick Prefix Beta-Lactamase Chloramphenicol Acetyl-Transferase Gram Positive Replication Protein puc9 Plasmid (E. coli rep origin) | 1,21 5187,5208 4120,4981 2176,2924 723,1742 399,649 | engineered_region feature/BioBrick engineered_region feature/BioBrick feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/misc |