Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Frank Sargent
Date created: 2016-10-10 11:00:00
Date modified: 2016-10-11 03:26:24
Truncated Colicin Ia Lacking Bacteriocin Active Domain
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1962002_sequence (Version 1) |
Description
Colicin Ia is a bacteriocin secreted by E. coli. Colicins typically have 3 domains, a translocation domain, a receptor binding domain, and a cytotoxic domain. If you are looking for a full-length Colicin Ia BioBrick it is located hereThis part encodes a truncated version of Colicin Ia that lacks the C-terminal bacteriocin domain. The part has an engineered multiple cloning site at the 3' end, preceding the RFC[10] suffix and regulation double stop codons, containing the following in-frame restriction sites: BamHI, KpnI, SalI, BglII and NheI. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin in a rapid and facile manner.
Notes
REC[10] compliantSource
We used the Chimera molecular modelling software to determine the sequence of the cytotoxic domain of Colicin iA, we then designed a truncated Colicin Ia, without this domain. The sequence was then codon optimised for E. coli K-12 and synthesised by IDT as a gBlock gene fragment. Using primers we amplified the truncated Colicin Ia with a multiple cloning site containing the following restriction sites: BamHI, KpnI, SalI, BglII and NheI located at the 3' end. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminal end of the truncated colicin and thereby generating synthetic colicins.| Sequence Annotation | Location | Component / Role(s) |
| Truncated Colicin Ia MCS | 1,1350 1351,1380 | CDS feature/cds feature/misc sequence_feature |