Coding

Part:BBa_K648013

Designed by: Jim Rose   Group: iGEM11_Penn_State   (2011-07-04)


GFP with Standard 25 Prefix/Suffix

This is the standard GFP protein reporter (E0040) cloned with the prefix/suffix required for standard 25 assembly. It can easily be used to create fusion proteins through this method.



Additional Tag

UCL iGEM 2017 used this BioBrick for its designs and noticed that this BioBrick actually contains the GFP protein reporter (E0040) fused to a FLAG epitope tag, followed by an enterokinase cleavage site. This is important if cell surface expression of this BioBrick is wished. Addition of enterokinase (also called enteropeptidase) to the solution allows easy testing of protein localisation (inside or outside of the membrane).



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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 641



Functional Parameters: Austin_UTexas

BBa_K648013 parameters

Burden Imposed by this Part:

Burden Value: 1.3 ± 1.4%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods, an explanation of the sources of burden, and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.

[edit]
Categories
//cds/reporter/gfp
//function/reporter/fluorescence
Parameters
colorGreen