Difference between revisions of "Part:BBa K2933274"

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<partinfo>BBa_K2933274 short</partinfo>
 
<partinfo>BBa_K2933274 short</partinfo>
  
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This part consists of RBS, protein coding sequence(His+Linker h+Sumo+Linker f+GIM-2) and T7 terminator,and the biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.
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This part consists of RBS, Linker h, protein coding sequence(His+Linker f+GIM-2) and T7 terminator,and the biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.
  
  
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===Usage and Biology===
 
===Usage and Biology===
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This composite part is made up with three basic parts(RBS, Linker h and T7 terminator)and a composite part(His+Linker f+GIM-2). It encodes a protein which is GIM-2 fused with His tag. The fusion protein is about 27.4 kD. It is convenient for us to purify our target protein.
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This composite part is made up with six basic parts(RBS , Linker h, His, Linker f ,  GIM-2 and T7 terminator ). It encodes a protein which is GIM-2 fused with His tag. The fusion protein is about 27.4 kD. It is convenient for us to purify our target protein.
  
 
===Molecular cloning===
 
===Molecular cloning===

Revision as of 14:17, 24 September 2019


RBS+Linker h+His+Linker f+GIM-2+T7 terminator

This part consists of RBS, Linker h, protein coding sequence(His+Linker f+GIM-2) and T7 terminator,and the biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 859
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 93
    Illegal NheI site found at 889
    Illegal PstI site found at 859
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 859
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 859
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

This composite part is made up with six basic parts(RBS , Linker h, His, Linker f , GIM-2 and T7 terminator ). It encodes a protein which is GIM-2 fused with His tag. The fusion protein is about 27.4 kD. It is convenient for us to purify our target protein.

Molecular cloning

GIM-2-PCR.jpeg
Figure 1. The result of PCR

References

1. Skagseth S , Akhter S , Paulsen M H , et al. Metallo-β-lactamase inhibitors by bioisosteric replacement: Preparation, activity and binding[J]. European Journal of Medicinal Chemistry, 2017, 135:159-173.

2. Wendel AF, MacKenzie CR. 2015. Characterization of a novel metallo-lactamase variant, GIM-2, from a clinical isolate of Enterobacter cloacae in Germany. Antimicrob Agents Chemother 59:1824 –1825.

3. Borra P S , Samuelsen O , Spencer J , et al. Crystal Structures of Pseudomonas aeruginosa GIM-1: Active-Site Plasticity in Metallo-beta-Lactamases[J]. Antimicrobial Agents and Chemotherapy, 2013, 57(2):848-854.

4. Susann S, Trine J C, Gro Elin K B, James S, Ørjan S, Hanna-Kirsti S. L. Role of Residues W228 and Y233 in the Structure and Activity of Metallo-β-Lactamase GIM-1. Antimicrobial Agents and Chemotherapy Jan 2016, 60 (2) 990-1002