Composite

Part:BBa_K3286100

Designed by: Robert Hooftman   Group: iGEM19_Wageningen_UR   (2019-10-14)
Revision as of 13:15, 21 October 2019 by Roberthooftman (Talk | contribs)

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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]


[edit]
Categories
Parameters
None