Si-tag1+3

BBa_K1592012 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1592012
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Shuyan Tang
Date created: 2015-09-06 11:00:00
Date modified: 2015-09-18 09:35:56

LIP prepro + E. coli ribosomal protein L2 (1-60,203-273aa) + YLcwp3 Fusion



Types
DnaRegion

Roles
CDS

Coding

Sequences BBa_K1592012_sequence (Version 1)

Description

__NOTOC__
BBa_K1592008 short

This is the cell display system of Yarrowia lipolytica, composed of LIP prepro, interest protein, and YLcwp3.LIP prepro is signal peptide used to secrete the interest protein out of the cell, and the YLcwp3 is the anchor domain binding the interest protein to the cell wall of yeast.We use this system to display silica-tag and test its binding characteristics.

Here we use this system to display Silica-tag on cell wall of Yarrowia lipolytica to binding the silica.

E.coli ribosomal protein L2 was found to bind tightly to silicon particles, which have surfaces that are oxidized to silica. This L2 silica-binding tag, called the 'Si-tag', can be used for one-step targeting of functional proteins on silica surfaces. The silica-binding domains of E. coli L2 was mapped to amino acids 1???60, 61-202 and 203???273, called Si-tag1, Si-tag2 and Si-tag3. We respectively test the silica-binding characteristics of this three regions and their combinations.

Notes

In order to replace the domains more conveniently, we respectivey added BamHI, SalI, NdeI between LIP2 prepro, E. coli ribosomal protein L2, GS linker and YLcwp3

Source

E. coli ribosomal protein L2 was synthesized by IDT, LIP2 prepro and YLcwp3 were cloning from the plasmid JMP62, and we fused this three sequences.

Sequence Annotation Location Component / Role(s)
LIP2 prepro
BamHI
Si-tag
SalI
GS Linker
NdeI
YLcwp3
1,99
100,105
106,501
502,507
508,552
553,558
559,924
CDS feature/protein
sequence_feature feature/misc
feature/protein CDS
sequence_feature feature/misc
feature/tag tag
sequence_feature feature/misc
CDS feature/protein
igem#experience
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igem#status
Planning
 
synbiohub#ownedBy
user/myers
 
synbiohub#ownedBy
user/james
 
synbiohub#topLevel
BBa_K1592012/1