Difference between revisions of "Part:BBa K2624005:Design"

 
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We checked the exact transcription start site of the hulc gene and replaced the sequence after it with the sequence that we designed so that, instead of the lncRNA, it would express the pri-miRNA analogue.
 
We checked the exact transcription start site of the hulc gene and replaced the sequence after it with the sequence that we designed so that, instead of the lncRNA, it would express the pri-miRNA analogue.
 
Then of course we added a SV40 poly(A) sequence that signals the end of a transcriptional unit.
 
Then of course we added a SV40 poly(A) sequence that signals the end of a transcriptional unit.
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In the end we added 5 different restriction sites for 5 different endonucleases, three in the front and two at the end, for recombination convenience.
 
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===Source===
 
===Source===
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===References===
 
===References===
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[1]Wang J, Liu X, Wu H, et al. CREB up-regulates long non-coding RNA, HULC expression through interaction with microRNA-372 in liver cancer[J]. Nucleic acids research, 2010, 38(16): 5366-5383.<br/>
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[2]Cheng H, Zhang Y, Wang H, et al. Regulation of MAP4K4 gene expression by RNA interference through an engineered theophylline-dependent hepatitis delta virus ribozyme switch[J]. Molecular BioSystems, 2016, 12(11): 3370-3376.<br/>
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[3]Han J, Lee Y, Yeom K H, et al. Molecular Basis for the Recognition of Primary microRNAs by the Drosha-DGCR8 Complex[J]. Cell, 2006, 125(5):887-901.

Latest revision as of 13:54, 17 August 2018


hulc-pri-miRNA(MAP4K4)


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 site found at 130


Design Notes

We checked the exact transcription start site of the hulc gene and replaced the sequence after it with the sequence that we designed so that, instead of the lncRNA, it would express the pri-miRNA analogue. Then of course we added a SV40 poly(A) sequence that signals the end of a transcriptional unit.

Source

Hulc promoter sequence comes from Swissregulon Database. The pri-miRNA analogue part is designed by us.

References

[1]Wang J, Liu X, Wu H, et al. CREB up-regulates long non-coding RNA, HULC expression through interaction with microRNA-372 in liver cancer[J]. Nucleic acids research, 2010, 38(16): 5366-5383.
[2]Cheng H, Zhang Y, Wang H, et al. Regulation of MAP4K4 gene expression by RNA interference through an engineered theophylline-dependent hepatitis delta virus ribozyme switch[J]. Molecular BioSystems, 2016, 12(11): 3370-3376.
[3]Han J, Lee Y, Yeom K H, et al. Molecular Basis for the Recognition of Primary microRNAs by the Drosha-DGCR8 Complex[J]. Cell, 2006, 125(5):887-901.