Plasmid

Part:BBa_K3179107:Design

Designed by: Jiaxian Xu   Group: iGEM19_SYSU-CHINA   (2019-10-15)


LSBR5n3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 83
    Illegal AgeI site found at 1547
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1376
    Illegal BsaI.rc site found at 2430
    Illegal SapI.rc site found at 758


Design Notes

This part is a part of measurement, which consists of mKate, miRNA target and EBFP and is put in the vector eEF1α. This plasmid has two eEF1α promoter, one promoter the transcribe of miRNA target and mKate, another promote the transcribe of EBFP. Because the efficiency of two promoter are similar, the transcription of mKate and EBFP’s mRNA are totally same. So, we can calculate the transcription by the expression of EBFP. Meanwhile, the mRNA of mKate has the miRNA target, so, we can get the amount of free miRNA by the ratio between EBFP and mKate which is also called the effective amount. In our design, this part is used to calculate the effective amount of three kinds of miRNA.

Source

LSBR5n3

References

1 Mohammadi P, Beerenwinkel N, Benenson Y. Automated design of synthetic cell classifier circuits using a two-step optimization strategy[J]. Cell systems, 2017, 4(2): 207-218. e14.