Difference between revisions of "Part:BBa J31008:Design"
 
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__NOTOC__
 
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<partinfo>BBa_J31008 short</partinfo>
 
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<partinfo>BBa_J31008 SequenceAndFeatures</partinfo>
 
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===Design Notes===
 
===Design Notes===
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RFP reverse was PCR amplified from mRFP (BBaE1010) using the following primers:
 
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5'-atgcactagtatggcttcctccgaagacgt "Spe mRFP F"
 
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5'-gcattctagattaagcaccggtggagtgac "Xba mRFP R"
 
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===Source===
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This part was PCR amplified from mRFP (<partinfo>BBa_E1010</partinfo>) using the following primers. The primers have non-annealing 5'- extensions that introduce a <font color='blue'>SpeI site</font> to the left and an <font color='red'>XbaI site</font> to the right of the coding region (allowing BioBrick cloning in the reverse orientation). Primer annealing sites are shown in bold.
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<br>Forward: 5’ ATGC<font color='blue'>ACTAGT</font><b>ATGGCTTCCTCCGAAGACGT</b>
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<br>Reverse: 5’ GCAT<font color='red'>TCTAGA</font><b>TTAAGCACCGGTGGAGTGAC</b>
  
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RFP reverse was PCR amplified from mRFP (BBaE1010).
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The final part was cloned into vector pSB1A3. The Biobricks on this part are not wild type, but the cut sites are still viable.  
  
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===References===
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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_J31008 Cloning Sites'''
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| style="background:lightgrey" |<font face="courier">5'--GAATTC GCGGCCGC T TCTAGA <font color='blue'>*</font> --RFP coding-- <font color='purple'>*</font> ACTAGT A GCGGCCG CTGCAG--<br>3'--CTTAAG CGCCGGCG A AGATCT <font color='blue'>*</font> -------------- <font color='purple'>*</font> TGATCA T CGCCGGC GACGTC--</font><br><br>
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'''Prefix'''<br>There is <font color='blue'>no G spacer (*)</font> between the XbaI and the Tet coding region.<br>
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'''Suffix'''<br>There is <font color='purple'>no T spacer (*)</font> between the Tet coding region and the SpeI site.
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|}

Latest revision as of 20:12, 15 February 2007

Design Notes

This part was PCR amplified from mRFP (BBa_E1010) using the following primers. The primers have non-annealing 5'- extensions that introduce a SpeI site to the left and an XbaI site to the right of the coding region (allowing BioBrick cloning in the reverse orientation). Primer annealing sites are shown in bold.
Forward: 5’ ATGCACTAGTATGGCTTCCTCCGAAGACGT
Reverse: 5’ GCATTCTAGATTAAGCACCGGTGGAGTGAC

The final part was cloned into vector pSB1A3. The Biobricks on this part are not wild type, but the cut 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_J31008 Cloning Sites 5'--GAATTC GCGGCCGC T TCTAGA * --RFP coding-- * ACTAGT A GCGGCCG CTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT * -------------- * TGATCA T CGCCGGC GACGTC--

Prefix
There is no G spacer (*) between the XbaI and the Tet coding region.
Suffix
There is no T spacer (*) between the Tet coding region and the SpeI site.