Tag

Part:BBa_K3589151

Designed by: Yannik Schermer   Group: iGEM20_TU_Kaiserslautern   (2020-10-24)
Revision as of 16:55, 25 October 2020 by Yanniks (Talk | contribs)


HA-RGS-8xHis tag for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the sequence of the HA-RGS-8His-tag (B5). Combined with a secretion signal of the Kaiser Collection from TU Kaiserslauterns 2019 iGEM Team like BBa_K3002007 (cCA (B2)), BBa_K3002008 (GLE (B2)) or BBa_3002009 (ARS (B2)), an appropriate promoter (BBa_K3002001 (PSAD promoter (A1-B1)) or BBa_K3002031 (PAR promoter (A1-B1))) and a terminator (e.g. BBa_K3002006 (RPL23 terminator)) your target protein should be secreted and you should be able to detect it via HA-antibody and purify it via a nickel-NTA column.
Note that this part misses a SP20-portion. The SP20-portion is reported to improve the secretion of proteins by increasing the glycosylation (see Team TU Kaiserslautern 2019's results). It is a repetition of Prolin and Serin, whereby Prolin is converted into hydroxyproline and then glycolyzed (Ramos-Martinez et. al, 2017).

BBa_K3589150 is a variation of this tag which possesses the SP20-portion.

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]


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Categories
Parameters
None