Difference between revisions of "Part:BBa K2758000:Design"

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===Source===
 
===Source===
  
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This sequence was constructed entirely with Biobricks from the iGEM registry. The sequences are: BBa_K346002 (mercury responsive promotor), BBa_B0030 (RBS), BBa_K1184000 (Killer Red), BBa_B0015 (double terminator).
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This sequence was constructed entirely with Biobricks from the iGEM registry. The sequences are: BBa_K346002 (mercury responsive promotor), BBa_B0030 (RBS), BBa_K1184000 (Killer Red), BBa_B0015 (double terminator). The part was designed using Snapgene and then sent to be synthesized by IDT. The biobrick was made by copying the sequences of the components from their respective registry pages in the order: BBa_K346002, BBa_B0030, BBa_K1184000, and BBa_B0015. An XbaI/SpeI ligation scar was added in between each part to simulate a ligation, however, we replaced the ligation scars flanking the RBS with HindIII and ApaI restriction sites. These restriction sites serve two purposes:
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1) To provide a method of checking whether or not the ligation of the biobrick to the PSB1C3 vector is successful
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2) To facilitate future characterization of the part with different RBS in order to delay or promote the expression of KR.
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https://static.igem.org/mediawiki/2018/5/5a/T--ColegioFDR_Peru--KRConstructSnapgene2018.png
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Figure 1. Map of the Killer Red construct displayed in the Snapgene software
  
 
===References===
 
===References===

Latest revision as of 01:21, 18 October 2018


Mercury activated Killer Red Protein suicide switch


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 237
    Illegal BsaI.rc site found at 528


Design Notes

This part is designed to work with another part that consists of a Hg accumulator. This part has been designed to express the Killer Red protein later than the accumulator is expressed from the other construct. In order to delay the expression of Killer Red protein relative to the accumulator, we inserted an RBS with moderate binding capabilities into the K2758000 construct. We built this construct through IDT adding the NotI restriction sites.

Source

This sequence was constructed entirely with Biobricks from the iGEM registry. The sequences are: BBa_K346002 (mercury responsive promotor), BBa_B0030 (RBS), BBa_K1184000 (Killer Red), BBa_B0015 (double terminator). The part was designed using Snapgene and then sent to be synthesized by IDT. The biobrick was made by copying the sequences of the components from their respective registry pages in the order: BBa_K346002, BBa_B0030, BBa_K1184000, and BBa_B0015. An XbaI/SpeI ligation scar was added in between each part to simulate a ligation, however, we replaced the ligation scars flanking the RBS with HindIII and ApaI restriction sites. These restriction sites serve two purposes:

1) To provide a method of checking whether or not the ligation of the biobrick to the PSB1C3 vector is successful
2) To facilitate future characterization of the part with different RBS in order to delay or promote the expression of KR.

T--ColegioFDR_Peru--KRConstructSnapgene2018.png Figure 1. Map of the Killer Red construct displayed in the Snapgene software

References