Difference between revisions of "Part:BBa K2940006"

 
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<partinfo>BBa_K2940006 short</partinfo>
 
<partinfo>BBa_K2940006 short</partinfo>
  
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Coding  Sequence of super foldin GFP with an N-terminal SpyCatcher
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Coding  Sequence of super fold GFP with an N-terminal SpyCatcher.
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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This part is used as a visual aid of the enzyme-SpyCatcher and curli fiber-SpyTag. The SpyCatcher is designed to be at the N-terminus of super fold GFP (sfGFP). A 2x GGGGS flexible linker was introduced between AzoR and SpyCatcher to make sure the domains of two proteins were not interfering with each other. SpyCatcher sequence and sfGFP sequence were acquired from iGEM registry (Part:BBa_K1650037 and Part:BBa_K515005).
  
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===Sequence and Features===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2940006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2940006 SequenceAndFeatures</partinfo>
  
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==Experimental Result==
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The assembled construct was transformed into BL21(DE3) strain to express recombinant protein. The colonies appeared to be green under uv light (Figure 1). The characterisation of this part need to be done in the future.
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<center>
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[[Image:T--Edinburgh OG--Wendy2.JPG|400px|Figure 1]]
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Figure 1. The GFP observation in transformation. Confirmed the activity of the sfGFP protein after linked to SpyCatcher.
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</center>
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 18:49, 21 October 2019


Coding Sequence of super folding GFP with an N-terminal SpyCatcher

Coding Sequence of super fold GFP with an N-terminal SpyCatcher.

Usage and Biology

This part is used as a visual aid of the enzyme-SpyCatcher and curli fiber-SpyTag. The SpyCatcher is designed to be at the N-terminus of super fold GFP (sfGFP). A 2x GGGGS flexible linker was introduced between AzoR and SpyCatcher to make sure the domains of two proteins were not interfering with each other. SpyCatcher sequence and sfGFP sequence were acquired from iGEM registry (Part:BBa_K1650037 and Part:BBa_K515005).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 222
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 592
  • 1000
    COMPATIBLE WITH RFC[1000]


Experimental Result

The assembled construct was transformed into BL21(DE3) strain to express recombinant protein. The colonies appeared to be green under uv light (Figure 1). The characterisation of this part need to be done in the future.

Figure 1

Figure 1. The GFP observation in transformation. Confirmed the activity of the sfGFP protein after linked to SpyCatcher.