Difference between revisions of "Part:BBa K3037001:Design"
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===Design Notes===
 
===Design Notes===
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Prescission site upstream and downstream.  
+
A PreScission recoginition site was added upstream and downstream of the coding region. This makes it possible to cleave the MBP tag off after purifictation. It can be specifically removed by digest with the PreScission Protease.
 +
This is very useful since this makes sure that the final pruified version of your protein of interest will not be influenced in its function or folding by this relatively large tag.
  
−
Biobrick used in RFC 25 standard
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To make it easy wo use this BioBrick for translational fusion we designed it in the RCF 25 BioBrick standart.
  
−
Codon optimized for <span style="font-style: italic;">E. coli</span> with all forbidden restriction enzyme sites removed
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Prefix: GAATTCGCGGCCGCTTCTAGATAAGGAGGTCAAAAATGgccggc
  
 +
Suffix: accggttaaTACTAGTAGCGGCCGCTGCAG
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Codon optimized for <span style="font-style: italic;">E. coli</span> with all forbidden restriction enzyme sites removed.
  
 
===Source===
 
===Source===

Latest revision as of 00:15, 19 October 2019


Maltose Binding Protein (MBP-tag)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 381
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 79


Design Notes

A PreScission recoginition site was added upstream and downstream of the coding region. This makes it possible to cleave the MBP tag off after purifictation. It can be specifically removed by digest with the PreScission Protease. This is very useful since this makes sure that the final pruified version of your protein of interest will not be influenced in its function or folding by this relatively large tag.

To make it easy wo use this BioBrick for translational fusion we designed it in the RCF 25 BioBrick standart.

Prefix: GAATTCGCGGCCGCTTCTAGATAAGGAGGTCAAAAATGgccggc

Suffix: accggttaaTACTAGTAGCGGCCGCTGCAG

Codon optimized for E. coli with all forbidden restriction enzyme sites removed.

Source

Synthesized by Integrated DNA Technologies (IDT)

References