BBa_K942000

BBa_K942000 Version 1

Component

This part has been discontinued.

Please consider using BBa_K942001 as a replacement.

Source:
http://parts.igem.org/Part:BBa_K942000
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_K942000_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 α-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 anti body.

Sequence Annotation Location Component / Role(s)
BBa_K942001
1,741
engineered_region feature/BioBrick
igem#experience
None
 
igem#sampleStatus
Discontinued
igem#status
Deleted
 
isReplacedBy
BBa_K942001/1
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K942000/1