Plasmid

Part:BBa_K3179100:Design

Designed by: Jiaxian Xu   Group: iGEM19_SYSU-CHINA   (2019-10-13)
Revision as of 21:57, 19 October 2019 by Bigxushen (Talk | contribs) (→‎Design Notes)


pTREK


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 195
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 201
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 857


Design Notes

This part is consisted of Vector pTRE, miR663b-target, 2kt-EGFP and miR885-5p-target. There is a CMV promoter on pTRE that can transcribe mRNAs with miR663b-target, 2kt-EGFP and miR885-5p-target. The translation of this mRNA is controlled by L7Ae and the amount of intracellular free miRNA663b and miRNA885-5p. When miRNA663b, miRNA885-5p and L7Ae exist, the translation of this mRNA is suppressed. The mRNA gene product is EGFP, and green fluorescence can be observed to facilitate observation of gene expression results. In our design, pTRE-miR663b-2kt-EGFP-miR885-5p is used for co-transduction with pTRE-L7Ae-miR592, and the compatibility and effectiveness of L7Ae-Kturn system and Tet-On system can be tested by measuring the expression amount of EGFP.

Source

pTRE-mi663b-2Kt-EGFP-miR885-5p

References