Translational_Unit

Part:BBa_K578005

Designed by: Kylie Standage-Beier   Group: iGEM11_Arizona_State   (2011-09-28)

J23102+E0840 (Constitutive GFP)

This is a combination of the GFP coding region (E0840), and the medium strength constitutive promoter, J23102.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 708

Confirmation of Function

BBa_K578005 was successfully subcloned into a variety of vectors, including pSB1C3, pSB1A3, pSB1K3, and pRSFDuet-1 (Novagen). The green fluorescent phenotype was observed. The image below show transforments of BL21(DE3) on the right and K12 MG1655 on the left, both with BBa_K578005 in pRSFDuet-1. Both show visible constitutive expression of GFP.

ASU GFP Duet.png

Expression in E.coli

These results indicate BBa_K578005 shows a high level of constitutive expression in both E.coli K12 MG1655 and E.coli BL21(DE3), allowing it to function as a visible marker for linked genetic elements, or as a visual screening tool for vector religation.


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