Difference between revisions of "Part:BBa J3101:Design"

 
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<partinfo>BBa_J3101 short</partinfo>
 
<partinfo>BBa_J3101 short</partinfo>
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The C > T point mutation occurred during cloning. It is located in the spacer region between the Fis binding sites; therefore, we believe that the mutation will not impact RE function. There is an insertion immediately  before the BioBrick suffix and several bases after the distal fis binding site that should affect RE function. RE is cloned in plasmid pSB1A2.
  
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The BioBrick prefix and suffix on this part are not wildtype but the restriction sites are still viable.
  
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===Source===
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{| width="800px" cellspacing="5"
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|- valign="top"
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| style="width:180px" | '''Standard BioBrick Cloning Sites''' (Knight)
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| style="background:lightgrey"|<font face="courier">5'--GAATTC GCGGCCGC T TCTAGA G ----insert---- T ACTAGT A GCGGCCG CTGCAG--<br>3'--CTTAAG CGCCGGCG A AGATCT C -------------- A TGATCA T CGCCGGC GACGTC--</font>
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|- valign="top"
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| style="width:180px" | '''BBa_J31001 Cloning Sites'''
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| style="background:lightgrey" |<font face="courier">5'--GAATTC GCGGCCGC T TCTAGA <font color='blue'>*</font> ------RE------ <font color='purple'>G</font> ACTAGT <font color='darkgreen'>T</font> GCGGCCG<font color='magenta'>C</font>CTGCAG--<br>3'--CTTAAG CGCCGGCG A AGATCT <font color='blue'>*</font> -------------- <font color='purple'>C</font> TGATCA <font color='darkgreen'>A</font> CGCCGGC<font color='magenta'>G</font>GACGTC--</font><br><br>
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'''Prefix'''<br>There is <font color='blue'>no G spacer (*)</font> between the XbaI and the RE.<br>
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'''Suffix'''<br>The T spacer between the RE and the SpeI site has changed to a <font color='purple'>G</font>.<br>The A spacer between the SpeI and the NotI has changed to a <font color='darkgreen'>T</font>.<br>There is an extra <font color='magenta'>C</font> between the NotI site and the PstI site
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|}
  
  
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===Source===
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The Recombination Enhancer sequence from ''Salmonella typhimurium''.
  
 
===References===
 
===References===
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<biblio>
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#Johnson pmid=2548848
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#Haykinson pmid=8508775
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#PerkinsBalding pmid=9244261
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</biblio>

Latest revision as of 09:34, 30 October 2008

Recombinational Enhancer (RE) for Hin/Hix inverting


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
    COMPATIBLE WITH RFC[1000]


Design Notes

The C > T point mutation occurred during cloning. It is located in the spacer region between the Fis binding sites; therefore, we believe that the mutation will not impact RE function. There is an insertion immediately before the BioBrick suffix and several bases after the distal fis binding site that should affect RE function. RE is cloned in plasmid pSB1A2.

The BioBrick prefix and suffix on this part are not wildtype but the restriction sites are still viable.

Standard BioBrick Cloning Sites (Knight) 5'--GAATTC GCGGCCGC T TCTAGA G ----insert---- T ACTAGT A GCGGCCG CTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT C -------------- A TGATCA T CGCCGGC GACGTC--
BBa_J31001 Cloning Sites 5'--GAATTC GCGGCCGC T TCTAGA * ------RE------ G ACTAGT T GCGGCCGCCTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT * -------------- C TGATCA A CGCCGGCGGACGTC--

Prefix
There is no G spacer (*) between the XbaI and the RE.
Suffix
The T spacer between the RE and the SpeI site has changed to a G.
The A spacer between the SpeI and the NotI has changed to a T.
There is an extra C between the NotI site and the PstI site


Source

The Recombination Enhancer sequence from Salmonella typhimurium.

References

<biblio>

  1. Johnson pmid=2548848
  2. Haykinson pmid=8508775
  3. PerkinsBalding pmid=9244261

</biblio>