Coding

Part:BBa_K1111012

Designed by: Sandra Chaudron, Caroline Desmurget and Mareike Apelt   Group: iGEM13_EPF_Lausanne   (2013-09-19)
Revision as of 17:59, 2 October 2013 by Sandelisa90 (Talk | contribs) (→‎Usage and Biology)

Ice Nucleation Protein fused to Streptavidin Alive

Introduction

What it is : INP sequence fused with Streptavidin Alive by gibson assembly. The INP-streptavidin construct expresses streptavidin at the outer membrane of E.Coli. INP is an outer membrane protein foud in the genome of Pseudomonas syringae. It is under the control of Lac promoter and thus is constitutively expressed in E.Coli. The expression of this plasmid was tested by transforming cells and looking for microscopy at either of a fluorescent antibody again streptavidin or a fluorescent biotin (biotin and streptavidin have a strong affinity, their Kd is 10-15 M).


Usage and Biology

This part was designed to express streptavidin on the outer membrane of E.coli in order to attach biotinylated nanoparticles to the cells. Thus, it can be used in any experiment involving biotin-streptavidin to put cargos on bacteria.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1727
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1649
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1727
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1727
    Illegal NgoMIV site found at 1036
    Illegal AgeI site found at 1691
    Illegal AgeI site found at 1742
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None