Difference between revisions of "Part:BBa K2933217"

 
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<partinfo>BBa_K2933217 parameters</partinfo>
 
<partinfo>BBa_K2933217 parameters</partinfo>
 
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==Usage and Biology===
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This composite part is made up with three basic parts, the His tag, T7 promoter, RBS and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of VIM-66 .  It is convenient for us to purify our target protein.<br>
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===Molecular cloning===
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We insert VIM-66 gene into the standard vector then transfer it into E.coli.
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  [[File:VIM-66-PCR.jpeg|600px|center|]] 
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<p style="text-align: center;">
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'''Figure 1.''' Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification

Revision as of 14:01, 21 September 2019


T7 promoter+RBS b+Linker h+His+Linker f+VIM-66+T7 terminator

This part consists of T7 promoter, RBS and protein coding sequence（His+Linker f+VIM-66），and the biological module can be built into E.coli for protein expression.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 47
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 169
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 942
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 47
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 47
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology=

This composite part is made up with three basic parts, the His tag, T7 promoter, RBS and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of VIM-66 . It is convenient for us to purify our target protein.

Molecular cloning

We insert VIM-66 gene into the standard vector then transfer it into E.coli.

VIM-66-PCR.jpeg

Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification