Part:BBa_K2267024:Design
SpyTag-GFP
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 866
Design Notes
A 6*His tag at the C end to facilitate purification. In order to increase the translation initiation rate, we also add an E.coil RBS.
As we have increase the yield of bacteria cellulose fulfiling the protection of the membrance will wake the BC functional Since the BC membrance is accepted due to its mechanical properties，we would like to introduce another system to add new characteries to our product.
According to the project of team Peking 2016 and resegrd papers we want to introduce the spytag-spychther system which is commonly used for binding. labeling immobilization and creating new kinds of application. We culture G.xylinus with the E.coli which can produce small peptid in the culture media evenly；thus， the spycather can be inlaid with BC membrance. The functional protein can be fused with spytag and be expressed bu the E.coli eddicieatly. We fuse the spytag with GFP.
Source
The SpyTag sequence comes from team Peking and sequence optimization is carried out by adding RBS and 6*His tag by PCR.