Difference between revisions of "Part:BBa K560015"

 
 
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<partinfo>BBa_K560015 short</partinfo>
 
<partinfo>BBa_K560015 short</partinfo>
  
 
The forth vector of the Golden gate series (adapted from psb1c3)  
 
The forth vector of the Golden gate series (adapted from psb1c3)  
This vector expedites golden gate cloning with BBa_K560012, BBa_K560013 and BBa_K560014.  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.  
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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_K560014 BBa_K560014].  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 13:28, 27 October 2011

BsaI Golden Gate Assembly No.4

The forth vector of the Golden gate series (adapted from psb1c3) This vector expedites golden gate cloning with BBa_K560012, BBa_K560013 and BBa_K560014. 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