BBa_K1989028

BBa_K1989028 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1989028
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yiming Dong
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-12 09:09:13

Triple spytag with mSA, His-tag , FlAsH-tag and signal peptide PhoA



Types
DnaRegion

Roles
CDS

Coding

Sequences BBa_K1989028_sequence (Version 1)

Description

The fused protein PhoA-Histag-3A-mSA-FlAsHtag possess both isopeptide bond forming function, secretion and collectable. Thus, using PhoA-Histag-3A- mSA -FlAsHtag as a part of hydrogel formation, we can obtain our multifunctional biomaterial.

Notes

The length of the linker between spytag may affect the physical and chemical property of hydrogel. As several attributes of our target proteins, especially their folding state in cells, we were unable to rationally design or choose a signal peptide for each. Nevertheless, we could build a Signal Peptide Repertory to screen for the best SP candidate. Our ultimate goal was to select the most appropriate ones from a huge amount of signal peptides.

Source

Inspiring from the self-catalysis of isopeptide bond between Lys and Asp in Streptococcus pyogenes fibronectin-binding protein FbaB, researchers split the catalytic domain and obtain two peptide called Spytag(the short one) and Spycatcher(the long one) which are able to form isopeptide bond with the other without any assistant. By fusing Spytag and Spycatcher with functional domains respectively, researchers solve the problem tactfully. In order to using Spytag and Spycatcher system as scaffold, we fused three Spytag spaced by (VPGVG)4 with 6xHistag in N-terminal and another functional protein called monomeric streptavidin(mSA) in C-terminal.

Sequence Annotation Location Component / Role(s)
Signal peptide PhoA
His-tag
Spytag-Spytag-Spytag
Linker
mSA
FlAsH-tag
24,90
91,129
130,377
382,461
466,824
832,870
feature/protein CDS
feature/tag tag
feature/protein CDS
CDS feature/protein
feature/protein CDS
feature/tag tag
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1989028/1