Part:BBa_K3589150

SP20-HA-RGS-8xHis tag for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the sequence of the SP20-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 efficiently and you should be able to detect it via HA-antibody and purify it via a nickel-NTA column.
The SP20 tag should improve the secretion of proteins by increasing the glycosylation. It is a repetition of Prolin and Serin, whereby Prolin is converted into hydroxyproline and then glycolyzed (Ramos-Martinez et. al, 2017).

Summary of the Results from Team Kaiserslautern 2020 for part BBa_K3589150

• We could not detect our secreted protein of interest when combined with this tag, as well as a cCA secretion signal.

For more information, please visit the registry pages for the L1 and L2 constructs containing this tag:

L1
BBa_K3589203
BBa_K3589204

L2
BBa_K3589209
BBa_K3589210

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]