Plasmid

Part:BBa_K3179104:Design

Designed by: Jiaxian Xu   Group: iGEM19_SYSU-CHINA   (2019-10-16)
Revision as of 04:24, 20 October 2019 by Bigxushen (Talk | contribs) (→‎Source)


hEF1a-rtTA3


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
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part consists with promoter hEF1α and basic part rtTA3 in vector pFUGW. hEF1α is a human elongation factor 1α gene promoter and is a strong mammalian promoter. hEFα is usually used in regular expression and has a stable expressional level, which is independent with cell type. rtTA is a key component of Tet-On system, and rtTA3 is third generation rtTA. rtTA3 is an activation factor of pTRE. rtTA3 isn’t active when it is present alone, and it can be active when combine with doxycycline (DOX), then the complex can combine to pTRE and start the downstream transcription. When DOX get removed the rtTA3 will became no activation again.

Source

hEF1a_rtTA3 was a gift from Ron Weiss (Addgene plasmid # 61472 ; http://n2t.net/addgene:61472 ; RRID:Addgene_61472)

References