Difference between revisions of "Part:BBa K560012"
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The first vector of the Golden gate series (adapted from psb1c3) | The first vector of the Golden gate series (adapted from psb1c3) | ||
| − | This vector expedites golden gate cloning with[https://parts.igem.org/wiki/index.php?title=Part:BBa_K560013 BBa_K560013], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K560014 BBa_K560014] 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. | + | This vector expedites golden gate cloning with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K560013 BBa_K560013], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K560014 BBa_K560014] 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. |
Latest revision as of 13:22, 27 October 2011
BsaI Golden Gate Assembly No.1
The first vector of the Golden gate series (adapted from psb1c3) This vector expedites golden gate cloning with BBa_K560013, BBa_K560014 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
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XbaI site found at 2073
Illegal PstI site found at 3 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal PstI site found at 3 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XhoI site found at 1041
Illegal XhoI site found at 1933 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XbaI site found at 2073
Illegal PstI site found at 3 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal XbaI site found at 2073
Illegal PstI site found at 3 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 2062
Illegal BsaI.rc site found at 14