Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Rachelle Varga
Date created: 2016-10-07 11:00:00
Date modified: 2016-10-08 08:20:16
pVeg->sec-TD1-KSCI-BBI-GFP
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K2008003_sequence (Version 1) |
Description
This part contains an active nonamer from the Bowman Birk Protease Inhibitor (BBI). A B. subtilis secretory signal peptide sequence is included to allow for secretion of the peptide directly into surrounding media. A transdermal tag (TD1, BBa_K1074000) is fused to the N-terminus of BBI to allow for diffusion of the peptide across the skin. Restriction enzyme sites BamHI and XmaI flank the BBI sequence for the interchangeability of protein product.This construct is fused to GFP (BBa_E0040) for visual detection, and the entire coding sequence is under the control of the constitutive B. subtilis promoter pVeg (BBa_K143012) and a strong B. subtilis RBS (BBa_K780001).
Note: This part is a composite part, not a basic part, but it could not be entered as composite at this time.
Notes
Bacillus subtilis-specific components were used (strong RBS and constitutive promoter) to produce high levels of expression of the BBI peptide fused to GFP for visualization. A secretory signal peptide sequence was included to allow for secretion of the peptide directly into surrounding media. A transdermal tag allowed for diffusion of the peptide across the skin. Internal restriction enzymes sites were added for peptide product interchangeability. Codons were optimized for B. subtilis.Source
Members of the lentil family produce the full BBI protein, and the genetic sequence for the peptide was sourced originally from genomic DNA.| Sequence Annotation | Location | Component / Role(s) |
| BBa_K143012 BBa_K780001 Start codon Sec tag BBa_K1074000 BBI with KSCI solubility tag BBa_E0040 Double stop codon | 1,97 98,113 123,125 126,209 210,242 243,248 285,995 996,1001 | feature/promoter promoter ribosome_entry_site feature/rbs start_codon feature/start feature/tag tag feature/tag tag feature/protein CDS feature/protein CDS feature/stop stop_codon |