Difference between revisions of "Part:BBa K2404000:Design"

 
 
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===Design Notes===
 
===Design Notes===
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To produce this protein we needed to create a gene construct with the TSH antagonist as the coding sequence. We used the Golden Gate cloning system to do this, and so needed to create upstream restriction enzyme recognition sites (i.e. Type IIS restriction endonucleases) that are suitable for Golden Gate reactions.
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We generated this part through the following steps:
  
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- We ordered the TSH gBlock from IDT that included 5' and 3' BsmBI restriction sites<br>
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- This was introduced into [https://parts.igem.org/Part:BBa_P10500 BBa_P10500] by digestion with BsmBI. This generated a 5' and 3' BsaI sites for level 1 cloning
  
  

Latest revision as of 21:00, 30 October 2017


A Thyroid Stimulating Hormone antagonist without His-Tags


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 540
    Illegal PstI site found at 582
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 540
    Illegal PstI site found at 582
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 607
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 540
    Illegal PstI site found at 582
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 540
    Illegal PstI site found at 582
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 794


Design Notes

We generated this part through the following steps:

- We ordered the TSH gBlock from IDT that included 5' and 3' BsmBI restriction sites

- This was introduced into BBa_P10500 by digestion with BsmBI. This generated a 5' and 3' BsaI sites for level 1 cloning


Source

Synthetic G-block. This modified protein was discovered in a paper by Fares et al in 2001.

References