Difference between revisions of "Part:BBa K2355302"

Revision as of 17:25, 26 October 2017

AmilCP-based composite part for insertion of cleavable peptide sequences in RFC[25]

This part was designed as an easy-to-use vector for evaluating the cleavage potential of different protein linkers in RFC[25] format. The device contains two major protein domains: A Single chain Avidin domain to bind the device to biotinylated surfaces, and AmilCP with his tag, a chromoprotein that acts as a reporter for cleavage of the inserted linker. The his tag allows for easy purification of protein. Between the two domains, NgoMIV and AgeI restriction sites are present, to allow for insertion of RFC[25] parts. Furthermore, between the two restriction sites of NgoMIV and AgeI, both a stop codon and frame shift is introduced, to prevent the expression of the AmilCP reporter if no linker is succesfully inserted.

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
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1711
Illegal AgeI site found at 1725
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1722

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2355302 short</partinfo>
-This part was designed as an easy-to-use vector for evaluating the cleavage potential of different protein linkers in RFC[25] format. The device contains two major protein domains: A Single chain Avidin domain to bind the device to biotinylated surfaces, and AmilCP, a chromoprotein that acts as a reporter for cleavage of the inserted linker. Between the two domains, NgoMIV and AgeI restriction sites are present, to allow for insertion of RFC[25] parts. Furthermore, between the two restriction sites of NgMIV and AgeI, both a stop codon and frame shift is introduced, to prevent the expression of the AmilCP reporter if no linker is succesfully inserted.
+This part was designed as an easy-to-use vector for evaluating the cleavage potential of different protein linkers in RFC[25] format. The device contains two major protein domains: A Single chain Avidin domain to bind the device to biotinylated surfaces, and AmilCP with his tag, a chromoprotein that acts as a reporter for cleavage of the inserted linker. The his tag allows for easy purification of protein.
+Between the two domains, NgoMIV and AgeI restriction sites are present, to allow for insertion of RFC[25] parts. Furthermore, between the two restriction sites of NgoMIV and AgeI, both a stop codon and frame shift is introduced, to prevent the expression of the AmilCP reporter if no linker is succesfully inserted.