Difference between revisions of "Part:BBa K3254001"

Revision as of 10:01, 9 October 2019

Int5attP-BsaI site-terminator-Int5attB

We expected that this part can be placed between a promoter and a translational unit part and work as a normally open (NO) switch for the downstreamed gene. and switch to ON state by excising the terminator ECK120010855 between the att sites when it reacted with Int5 integrase (BBa_K3254006). However, we haven't observed the expression of downstreamed gfp gene. That's may caused by the potential terminator activity of attL site after the recombination. Two BsaI restriction site were added between attB site and terminator. As a result, it can work as a normally closed (NC) switch for the gene which was inserted between the two BsaI site and switch to OFF state when it was excised.

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 site found at 74
Illegal BsaI.rc site found at 62

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3254001 short</partinfo>
-We expected that this part can be placed between a promoter and a translational unit part and work as a normally open (NO) switch for the downstreamed gene. and switch to ON state by excising the terminator ECK120010855 between the att sites when it reacted with Int5 integrase (BBa_K3254006). However, we haven't observed the expression of downstreamed gfp gene. That's may caused by the potential terminator activity of attL site after the recombination.
+We expected that this part can be placed between a promoter and a translational unit part and work as a normally open (NO) switch for the downstreamed gene. and switch to ON state by excising the terminator ECK120010855 between the att sites when it reacted with Int5 integrase ([[Part:BBa_K3254006|BBa_K3254006]]). However, we haven't observed the expression of downstreamed gfp gene. That's may caused by the potential terminator activity of attL site after the recombination.
 Two BsaI restriction site were added between attB site and terminator. As a result, it can work as a normally closed (NC) switch for the gene which was inserted between the two BsaI site and switch to OFF state when it was excised.