Difference between revisions of "Part:BBa K5387002:Design"

 
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===Design Notes===
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Some parts of the protein are membrane bound, which makes them much harder to solve and express in E. coli. We therefore tried various different primers to try to clone different parts of the protein to pET28a(+), listed below:
  
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__NOTOC__
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To clone the whole sequence into pET28a(+) between the NdeI and EcoRI restriction sites:
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<partinfo>BBa_K5387002 short</partinfo>
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: fwd: 5'-GCGCCATATGATGGACGGACCCAGGTCAG-3'
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: rev: 5'-GCGCGAATTCTTAGGCGCCACCTCTGCTTGCCATC-3'
  
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<partinfo>BBa_K5387002 SequenceAndFeatures</partinfo>
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To delete the first 100 amino acids:
 +
: fwd: 5'-GCGCCATATGGGTATGCTTGTGGTCAACGG-3'
 +
: rev: 5'-GCGCGAATTCTTAGGCGCCACCTCTGCTTGCCATC-3'
  
 +
To delete the last 30 amino acids:
 +
: fwd: 5'-GCGCCATATGATGGACGGACCCAGGTCAG-3'
 +
: rev: 5'-CAGGAGCCACGTCAACCTGAATAACACC-3'
  
−
===Design Notes===
+
To delete the first 100 and last 30 and amino acids:
−
Some parts of the protein are membrane bound, which makes them much harder to solve and express in E. coli. We therefore tried various different primers to try to clone different parts of the protein to pET28a(+), listed below:
+
: fwd: 5'-GCGCCATATGGGTATGCTTGTGGTCAACGG-3'
−
 
+
: rev: 5'-CAGGAGCCACGTCAACCTGAATAACACC-3'
  
  
 
===Source===
 
===Source===
−
 
 
The genes is expressed in human cells.
 
The genes is expressed in human cells.
  
 
===References===
 
===References===

Revision as of 12:46, 20 August 2024

Design Notes

Some parts of the protein are membrane bound, which makes them much harder to solve and express in E. coli. We therefore tried various different primers to try to clone different parts of the protein to pET28a(+), listed below:

To clone the whole sequence into pET28a(+) between the NdeI and EcoRI restriction sites:

fwd: 5'-GCGCCATATGATGGACGGACCCAGGTCAG-3'
rev: 5'-GCGCGAATTCTTAGGCGCCACCTCTGCTTGCCATC-3'

To delete the first 100 amino acids:

fwd: 5'-GCGCCATATGGGTATGCTTGTGGTCAACGG-3'
rev: 5'-GCGCGAATTCTTAGGCGCCACCTCTGCTTGCCATC-3'

To delete the last 30 amino acids:

fwd: 5'-GCGCCATATGATGGACGGACCCAGGTCAG-3'
rev: 5'-CAGGAGCCACGTCAACCTGAATAACACC-3'

To delete the first 100 and last 30 and amino acids:

fwd: 5'-GCGCCATATGGGTATGCTTGTGGTCAACGG-3'
rev: 5'-CAGGAGCCACGTCAACCTGAATAACACC-3'


Source

The genes is expressed in human cells.

References