Part:BBa_K1943019

TetOn

We submitted an inducible expression systems, which we constructed to regulate the expression of downstream gene in CHO K1 cell line.

TRE-3G promoter is a tetracycline-responsive promoter. The expression of downstream gene can be induced by the addition of doxycycline (DOX). It has been submitted by our predecessor, SUSTC-Shenzhen in 2014 (see the biobrick BBa_K1431301 ) They combined these 2 parts into one plasmid. It is useful to improve the success rate of transfection. Considering the difficulty to cut the whole fragment as a submitted part, they only constructed the assembly with a part of the regulation system.

The other part of the system includes a promoter SV40 and the tetracycline (Tet)-regulated transactivator Tet-On 3G. This year we submitted this part connected with 2A and pure gene, the insertion can be confirmed by antibiotic selection, using 3μg/mL puromycin.

The use of this part will be wide. Following teams can insert the any target gene after promoter TRE 3G to make it under the control of doxycycline. The regulate part (SV40 to TetON 3G) and the response part (pTRE and target gene) could be constructed into 2 separated plasmid or the same plasmid as we did.

figure: the application of dox on transcription

Tetracycline-responsive promoter, expression induced by doxycycline.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 857