Difference between revisions of "Part:BBa K1993035"

 
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<partinfo>BBa_K1993035 short</partinfo>
 
<partinfo>BBa_K1993035 short</partinfo>
  
IRES-Luciferase-T2A-dtomato-T2A-hFTH
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In order to assemble our device easily since we had constructed plasmids by Gateway technology, we constructed a composite part transcripting IRES and Luciferase-T2A-dTomato-T2A-eGFP which is convenient for running Gateway BP and LR reactions.
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We introduced Internal ribosome entry site (IRES) (Details can be seen from [https://parts.igem.org/Part:BBa_K1993016 BBa_K1993016]) to ensure the translating initiation in an end-independent manner, as part of the greater process of protein synthesis.
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Firefly(Photinus pyralis) Luciferase (Details can be seen from [https://parts.igem.org/Part:BBa_K1993018 BBa_K1993018]) is a kind of oxidative enzyme that produce bioluminescence. It is able to oxidize luciferin and produce detectable bioluminescence. It’s convenient to observe biological processes, especially allowing for observation of cells non-invasively. So we added such a gene to ensure observation in vivo.
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To expand the function of our composite part, we added two more genes: dTomato(Details can be seen from [https://parts.igem.org/Part:BBa_K1993020 BBa_K1993020])  used for observation in vitro conveniently and easily; hFTH(Details can be seen from [https://parts.igem.org/Part:BBa_K1993021 BBa_K1993021]), another protein that ccould be observed in vivo by MRI and applied to human body. What’s more, in order to make sure the expression of all three proteins, we added a T2A gene(Details can be seen from [https://parts.igem.org/Part:BBa_K1993019 BBa_K1993019]) between every two protein coding sequences. By constructing such a composite part, we could run our Gateway BP and LR reactions more easily through adding the Gateway loci to both end of that composite part.
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Revision as of 13:09, 18 October 2016


IRES-Luciferase-T2A-dtomato-T2A-hFTH

In order to assemble our device easily since we had constructed plasmids by Gateway technology, we constructed a composite part transcripting IRES and Luciferase-T2A-dTomato-T2A-eGFP which is convenient for running Gateway BP and LR reactions. We introduced Internal ribosome entry site (IRES) (Details can be seen from BBa_K1993016) to ensure the translating initiation in an end-independent manner, as part of the greater process of protein synthesis. Firefly(Photinus pyralis) Luciferase (Details can be seen from BBa_K1993018) is a kind of oxidative enzyme that produce bioluminescence. It is able to oxidize luciferin and produce detectable bioluminescence. It’s convenient to observe biological processes, especially allowing for observation of cells non-invasively. So we added such a gene to ensure observation in vivo. To expand the function of our composite part, we added two more genes: dTomato(Details can be seen from BBa_K1993020) used for observation in vitro conveniently and easily; hFTH(Details can be seen from BBa_K1993021), another protein that ccould be observed in vivo by MRI and applied to human body. What’s more, in order to make sure the expression of all three proteins, we added a T2A gene(Details can be seen from BBa_K1993019) between every two protein coding sequences. By constructing such a composite part, we could run our Gateway BP and LR reactions more easily through adding the Gateway loci to both end of that composite part.


Sequence and Features


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 SapI.rc site found at 1396