Team:Cambridge/Experiments/Plasmid Constructs

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=Constructs=
=Constructs=
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As part of our project we designed a library of reflectin expression plasmids, as well as constructs with reflectin targetted for export to the periplasm, and finally plasmids with reflectin transcriptional fusions with GFP. Diagrams of all of them are given below.  
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As part of our project we designed a library of reflectin expression plasmids, as well as constructs with reflectin transcriptional fusions with GFP, and finally plasmids with reflectin targetted for export to the periplasm. Diagrams of all of them are given below.  
   
   
==Expression Constructs==
==Expression Constructs==
===GA1===
===GA1===
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contents
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Reflectin A1 is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies).
[[File:cam_igem_map_GA1.jpg| 700px]]
[[File:cam_igem_map_GA1.jpg| 700px]]
===GA2===
===GA2===
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Reflectin A1 is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies).
[[File:cam_igem_map_GA2.jpg | 700px]]
[[File:cam_igem_map_GA2.jpg | 700px]]
===GA3===
===GA3===
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Reflectin A1 with a HIS tag at the N-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies).
[[File:cam_igem_map_GA3.jpg | 700px]]
[[File:cam_igem_map_GA3.jpg | 700px]]
===GA4===
===GA4===
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Reflectin A1 with a HIS tag at the N-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies).
[[File:cam_igem_map_GA4.jpg | 700px]]
[[File:cam_igem_map_GA4.jpg | 700px]]
===GA5===
===GA5===
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Reflectin A2 is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies).
[[File:cam_igem_map_GA5.jpg | 700px]]
[[File:cam_igem_map_GA5.jpg | 700px]]
===GA6===
===GA6===
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Reflectin 1B is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies).
[[File:cam_igem_map_GA6.jpg | 700px]]
[[File:cam_igem_map_GA6.jpg | 700px]]
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==Export Constructs==
 
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===GA17===
 
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tat export construct
 
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[[File:cam_igem_map_GA17.jpg |center | 700px ]]
 
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===GA18===
 
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[[File:cam_igem_map_GA18.jpg |center | 700px ]]
 
==GFP fusions==
==GFP fusions==
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===GA13===
===GA13===
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Reflectin A1 fused to GFP at the C-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies).
[[File:cam_igem_map_GA13.jpg | 700px]]
[[File:cam_igem_map_GA13.jpg | 700px]]
===GA14===
===GA14===
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Reflectin A1 fused to GFP at the C-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies).
[[File:cam_igem_map_GA14.jpg | 700px]]
[[File:cam_igem_map_GA14.jpg | 700px]]
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 +
==Export Constructs==
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 +
===GA17===
 +
Reflectin A1 is fused to the TorA signal peptide at the N-terminus, and his-tagged at the C-terminus. It is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). TorA signal targets the protein to the Tat export pathway in E.coli.
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[[File:cam_igem_map_GA17.jpg |center | 700px ]]
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 +
===GA18===
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Reflectin A1 is fused to the TorA signal peptide at the N-terminus, and fused to GFP at the C-terminus. It is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). TorA signal targets the protein to the Tat export pathway in E.coli.
 +
[[File:cam_igem_map_GA18.jpg |center | 700px ]]
{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}
{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}

Revision as of 16:39, 20 September 2011

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Contents

Constructs

As part of our project we designed a library of reflectin expression plasmids, as well as constructs with reflectin transcriptional fusions with GFP, and finally plasmids with reflectin targetted for export to the periplasm. Diagrams of all of them are given below.

Expression Constructs

GA1

Reflectin A1 is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). Cam igem map GA1.jpg

GA2

Reflectin A1 is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies). Cam igem map GA2.jpg

GA3

Reflectin A1 with a HIS tag at the N-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). Cam igem map GA3.jpg

GA4

Reflectin A1 with a HIS tag at the N-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies). Cam igem map GA4.jpg

GA5

Reflectin A2 is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies). Cam igem map GA5.jpg

GA6

Reflectin 1B is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies). Cam igem map GA6.jpg


GFP fusions

GA13

Reflectin A1 fused to GFP at the C-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). Cam igem map GA13.jpg

GA14

Reflectin A1 fused to GFP at the C-terminus is expressed under the control of the arabinose-inducible pBAD promoter on a low copy number plasmid pSB3K3 (10-12 copies). Cam igem map GA14.jpg


Export Constructs

GA17

Reflectin A1 is fused to the TorA signal peptide at the N-terminus, and his-tagged at the C-terminus. It is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). TorA signal targets the protein to the Tat export pathway in E.coli.

Cam igem map GA17.jpg

GA18

Reflectin A1 is fused to the TorA signal peptide at the N-terminus, and fused to GFP at the C-terminus. It is expressed under the control of the arabinose-inducible pBAD promoter on a high copy number plasmid pSB1A3 (100-300 copies). TorA signal targets the protein to the Tat export pathway in E.coli.

Cam igem map GA18.jpg