Difference between revisions of "Part:BBa K1321009"

 
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<partinfo>BBa_K1321009 short</partinfo>
 
<partinfo>BBa_K1321009 short</partinfo>
  
Nickel Binding Protein in Freiburg format (RFC 25) to allow for easy use in fusion proteins.
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'''**Important note**''': Please be aware we discovered that this part is out of frame, leading to an incorrect translation and read-through of the stop-codon. This also means any subsequent fusion proteins to the C-terminal are out of frame. This is because there is a deletion of a base compared to the equivalent coding sequence from ''Heliobacter pylori'' (GenBank CP000012.1 bp1391851-1392033, reverse complement strand). This existed already in BBa_K1151001 from which we cloned our part. To correct the frame-shift, a 'C' needs to be added between bases 49 and 50.
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This part is BBa_K1151001 in Frieburg (RFC[25]) format to allow for easy use in fusion proteins. Note that the start and stop codon, plus 6 bp either side of the sequence, are included the RFC25 prefix and suffix which is not shown. It is a histidine-rich metal binding protein which binds up to 5 Ni2+ ions (please see the original BBa_K1151001 biobrick page from team Università del Salento Lecce 2013 for more detailed information).
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For expression purposes this part is available with T7 promoter (BBa_K1321163) or LacI promoter (BBa_K1321162). We used this part to create a [http://2014.igem.org/Team:Imperial/Functionalisation library of fusion proteins with cellulose-binding domains].
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===Usage and Biology===
 
===Usage and Biology===
 
 
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<span class='h3bb'>Sequence and Features</span>
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===Sequence and Features===
 
<partinfo>BBa_K1321009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1321009 SequenceAndFeatures</partinfo>
  

Latest revision as of 01:37, 17 October 2014

Nickel Binding Protein (NiBP) Freiburg format (RFC[25])

**Important note**: Please be aware we discovered that this part is out of frame, leading to an incorrect translation and read-through of the stop-codon. This also means any subsequent fusion proteins to the C-terminal are out of frame. This is because there is a deletion of a base compared to the equivalent coding sequence from Heliobacter pylori (GenBank CP000012.1 bp1391851-1392033, reverse complement strand). This existed already in BBa_K1151001 from which we cloned our part. To correct the frame-shift, a 'C' needs to be added between bases 49 and 50.


This part is BBa_K1151001 in Frieburg (RFC[25]) format to allow for easy use in fusion proteins. Note that the start and stop codon, plus 6 bp either side of the sequence, are included the RFC25 prefix and suffix which is not shown. It is a histidine-rich metal binding protein which binds up to 5 Ni2+ ions (please see the original BBa_K1151001 biobrick page from team Università del Salento Lecce 2013 for more detailed information).

For expression purposes this part is available with T7 promoter (BBa_K1321163) or LacI promoter (BBa_K1321162). We used this part to create a [http://2014.igem.org/Team:Imperial/Functionalisation library of fusion proteins with cellulose-binding domains].


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]