Coding
NdeI+E0040

Part:BBa_K1819000

Designed by: Lais Ribovski   Group: iGEM15_Brasil-USP   (2015-09-13)

Designed by Brasil-USP team, BBa_K1819000 is a GFP coding sequence with an N-terminal linker, which is 5’-flanked by NdeI restriction site.

It allows to characterize protein expression and secretion and also may serve to other IGEM teams that would like to express a chimera C-terminally attached to GFP, which exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range.

We also added GGGS amino acid twice as a spacer to avoid interaction between two consecutive proteins, allowing proper folding. This strategy was already applied to BBa_K1489002 part.


Figure 1 - Schematic representation of BBa_K1819000 insert


Characterization


Figure 2 - Gel restriction analysis


We added this part as an improvement to BBa_E0040, a green fluorescente protein, for protein fusion (see Experience page) .
This part was characterized using two constitutive promoters from Anderson library (BBa_J23106 and BBa_J23117). See BBa_K1819006 and BBa_K1819007 pages for more information.


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 674
[edit]
Categories
//chassis/prokaryote/ecoli
//function/reporter
//function/reporter/fluorescence
Parameters
colorGreen
directionForward
emission
emit511
excitation
excite501
kegg
lum
proteinGFPmut3b
swisspro