Composite

Part:BBa_K1431301

Designed by: Rifei Chen, Yushan Zhang   Group: iGEM14_SUSTC-Shenzhen   (2014-10-10)
Revision as of 13:29, 17 October 2014 by ChenRifei (Talk | contribs)

TRE-3G promoter+SV40 PolyA, an ideal controller of mammalian gene expression with Tet-On 3G protein

TRE-3G promoter is a kind of eukaryocyte cell promoters that will be switched on after combine with the compound of Tet-On 3G(BBa_K1431101) protein and doxycycline(Dox) mix.
The inducible promoter PTRE3G provides for very low basal expression and high maximal expression after induction (Loew et. al., 2010). It consists of 7 repeats of a 19 bp tet operator sequence located upstream of a minimal CMV promoter. In the presence of Dox, Tet-On 3G binds specifically to PTRE3G and activates transcription of the downstream GOI. PTRE3G lacks binding sites for endogenous mammalian transcription factors, so it is virtually silent in the absence of induction. That is a very useful way to control whether a protein express or not.

There are two BbsI sites between promoter and PolyA which can make the coding sequence seamless gather with them. So we can also test how many nucleotides interval between promoter and coding sequence will make the highest expression efficiency.

Sequence and Features


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
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None