Difference between revisions of "Part:BBa K560014"

Latest revision as of 13:26, 27 October 2011

BsaI Golden Gate Assembly No.3

The third vector of the Golden gate series (adapted from psb1c3) This vector expedites golden gate cloning with BBa_K560012, BBa_K560013 and BBa_K560015. To make this set of vector compatible with the standard biobricks, we have maintained PstI and XbaI recognition sites. Flanking these two sites are the cleavage and recognition sites of BsaI, a kind of Type IIS restriction endonuclease. This kind of restriction endonucleases cleaves outside their recognition sites thus creating different sticky ends. If you want to assemble four biobricks at a time, you can put the biobricks in order into corresponding vector by the digestion of PstI and Xbal. Then mix these assembled vectors and a backbone (PFUS4 from TALEN golden gate kit). Under the effect of ligase and Bsal, these biobricks will have sticky ends nose to tails and ligate in order in a single reaction.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XbaI site found at 2073
Illegal PstI site found at 3
12
INCOMPATIBLE WITH RFC[12]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal PstI site found at 3
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XhoI site found at 1041
Illegal XhoI site found at 1933
23
INCOMPATIBLE WITH RFC[23]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XbaI site found at 2073
Illegal PstI site found at 3
25
INCOMPATIBLE WITH RFC[25]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XbaI site found at 2073
Illegal PstI site found at 3
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 2062
Illegal BsaI.rc site found at 14

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K560014 short</partinfo>
 The third vector of the Golden gate series (adapted from psb1c3)
-This vector expedites golden gate cloning with BBa_K560012, BBa_K560013 and BBa_K560015.  To make this set of vector compatible with the standard biobricks, we have maintained PstI and XbaI recognition sites. Flanking these two sites are the cleavage and recognition sites of BsaI, a kind of Type IIS restriction endonuclease. This kind of restriction endonucleases cleaves outside their recognition sites thus creating different sticky ends. If you want to assemble four biobricks at a time, you can put the biobricks in order into corresponding vector by the digestion of PstI and Xbal. Then mix these assembled vectors and a backbone (PFUS4 from TALEN golden gate kit). Under the effect of ligase and Bsal, these biobricks will have sticky ends nose to tails and ligate in order in a single reaction.
+This vector expedites golden gate cloning with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K560012 BBa_K560012], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K560013 BBa_K560013] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K560015 BBa_K560015].  To make this set of vector compatible with the standard biobricks, we have maintained PstI and XbaI recognition sites. Flanking these two sites are the cleavage and recognition sites of BsaI, a kind of Type IIS restriction endonuclease. This kind of restriction endonucleases cleaves outside their recognition sites thus creating different sticky ends. If you want to assemble four biobricks at a time, you can put the biobricks in order into corresponding vector by the digestion of PstI and Xbal. Then mix these assembled vectors and a backbone (PFUS4 from TALEN golden gate kit). Under the effect of ligase and Bsal, these biobricks will have sticky ends nose to tails and ligate in order in a single reaction.