Coding

Part:BBa_K209403

Designed by: Katja Kolar   Group: iGEM09_UCSF   (2009-10-02)

AarI A-B part, signal sequence-FLAG

GPCR N terminal tags flanked by AarI AB sites.

Usage and Biology

Intended to be used as an N-terminal addition to G protein-coupled receptors. The signal sequence from prolactin is reported to increase surface expression of GPCRs and is cleaved off leaving the free FLAG epitope, which is recognized by the M1 antibody.

Documentation and characterization

We did not conduct a specific comparison of GPCR expression with and without the signal sequence. However, the signal sequence was used for a number of receptors we tested in our GPCR screen, described here: http://2009.igem.org/Team:UCSF/Navigation


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 128
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 84


[edit]
Categories
Parameters
n/aAarI A-B part, signal sequence-FLAG