Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Luis Mario Leal Garza
Date created: 2012-09-25 11:00:00
Date modified: 2015-05-08 01:13:48
SS+antiHis+Etag
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K942015_sequence (Version 1) |
Description
This part is a single chain variable fragment (scFv), this is a fusion of the variable domains of antibodies. The antigen this scFv binds to with a greater stability is a C-terminal 6xHis tag; nevertheless it binds to tags with as less as 3xHis. It is formed by two domains, the variable heavy chain domain (VH) and the variable light chain domain (VL) (Kaufmann, Lindner, & Honegger, 2002).This protein has an estimated weight of 27.4kDa. It binds with the strongest force to C-terminal 6x His tag, but binds also to other types of His tags with different binding strength. The E-tag at the C-terminal for purification was not used in the end, instead an ultrafiltration with a 10kDa MWCO membrane was used. (Gasteiger E., 2005)
About Safety
The part itself does not present any biohazard characteristic, but it is important to remember that it is designed to work in Pichia pastoris and that a biosecurity chamber is recommended to handle this organism. Its secretion to the media should not suppose any problem.
Notes
This part is codon-optimized to be expressed in yeast. It is flanked by a secretion signal (SS) from the alpha-mating factor pre-sequence which makes the protein secretable by the yeast. An Etag was added at the C-terminal for affinity purification by L protein. This protein can be combined in fusion with another by a linker (GGGGS)4 in the C-terminal.Source
This part is a single chain variable fragment (scFv), this is a fusion of the variable domains of antibodies, its domains come from a murine antibody.| Sequence Annotation | Location | Component / Role(s) |
| Secretion signal alfa mating factor presequence E tag | 1,57 798,837 | feature/misc sequence_feature feature/tag tag |