Coding

Part:BBa_K4139015

Designed by: Zachary Robinson & Jason Luddu   Group: iGEM22_Lethbridge   (2023-09-26)


COAPE - GFP

This construct starts with an OmpA tag and is a signaling peptide that will bring the protein from cytoplasm to the periplasm. This is followed by enhanced green fluorescent protein (eGFP) then our Chimeric Anti-PbEL04 (CAPE). The CAPE proportion is the same as our CAPE protein. Finally we added a His-tag to further help with purification. The function of this protein is to bind to PbEL04 and glow once binded correctly. The reason for using eGFP is to show if proper binding has occurred. If proper binding has occurred then the protein will glow green when blue light in the 470 nm to 498 nm range. The difference between CAPE-GFP (BBa_K4139013) and COAPE-GFP (BBa_K4139015) is the OmpA tag which was used as a solubility tag. This construct was successfully purified by our team.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 946
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 946
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 946
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 946
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/prokaryote/ecoli
//function/reporter/fluorescence
Parameters
directionForward
emission510 nm
proteinPbEL04
tagOmpA & His-Tag