Part:BBa_K3286100

Expression construct for proteins in E. coli BL21DE3

This construct can be used to produce high amounts of protein in E. coli BL21DE3 (or another strain containing a T7 polymerase BBa_K1614000). Due to the Bicistronic Design (BCD) system BBa_M36516 [1], protein production is increased. Recombinant protein can be isolated using the Strep tag (BBa_K3286101), which can be removed with TEV (Tomato Etch Virus) protease at the TEV protease recognition site (BBa_K2927004). Integrate the protein of interest directly after the strep tag and make sure it is in frame. Also, remove its start codon.

[1] V. K. Mutalik et al., “Precise and reliable gene expression via standard transcription and translation initiation elements,” Nat. Methods, vol. 10, no. 4, pp. 354–360, Apr. 2013.

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]