Plasmid

Part:BBa_K3179106:Design

Designed by: Jiaxian Xu   Group: iGEM19_SYSU-CHINA   (2019-10-16)
Revision as of 22:27, 20 October 2019 by Bigxushen (Talk | contribs) (→‎References)

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pT2mK2E


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1750
    Illegal XbaI site found at 4374
    Illegal PstI site found at 2500
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2500
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3326
    Illegal BamHI site found at 954
    Illegal BamHI site found at 3578
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1750
    Illegal XbaI site found at 4374
    Illegal PstI site found at 2500
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1750
    Illegal XbaI site found at 4374
    Illegal PstI site found at 2500
    Illegal NgoMIV site found at 1224
    Illegal NgoMIV site found at 2219
    Illegal NgoMIV site found at 3848
    Illegal AgeI site found at 960
    Illegal AgeI site found at 3584
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3352


Design Notes

This part consists of miR663b-target,2kt-E1A, E1B 55K and miR885-5p-target, which are put in the vector pTRE. From promoter CMV our plasmid can transcribe mRNA with miR663b-target, 2kt-E1A, E1B 55K and miR885-5p-target, whose translation can be regulated by level of free miRNA663b and miRNA885-5p, and also by L7Ae. When they exist, the translation of this mRNA will be suppressed. The production is E1A and E1B 55K, E1A can start the expression of adenoviral gene, and E1B 55K can enhance the ability of virus to lyse the cell. During assembling, we use P2A sequence link E1A and E1B 55K, which can maintain the expression level of two gene when sharing one promoter.

Source

pTRE-mi663b-2Kt-E1-E1B55K-miR885-5p

References

1 Wroblewska L, Kitada T, Endo K, et al. Mammalian synthetic circuits with RNA binding proteins for RNA-only delivery[J]. Nature biotechnology, 2015, 33(8): 839.
2 Berk A J. Functions of adenovirus E1A[J]. Cancer surveys, 1986, 5(2): 367-387.
3 Martin M E D, Berk A J. Adenovirus E1B 55K represses p53 activation in vitro[J]. Journal of virology, 1998, 72(4): 3146-3154.
4 Kim J H, Lee S R, Li L H, et al. High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice[J]. PloS one, 2011, 6(4): e18556.