Composite

Part:BBa_K3286100

Designed by: Robert Hooftman   Group: iGEM19_Wageningen_UR   (2019-10-14)
Revision as of 16:27, 20 October 2019 by SJK (Talk | contribs)


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). Due to the Bicistronic Design (BCD) system [1], protein production is increased. Recombinant protein can be isolated using the Strep tag, which can be removed with TEV (Tomato Etch Virus) protease at the TEV protease recognition site. 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]


[edit]
Categories
Parameters
None