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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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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===Source===
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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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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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RFP reverse was PCR amplified from mRFP (BBaE1010).
  
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{| width="800px" cellspacing="5"
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===References===
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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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|}
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Revision as of 18:08, 15 February 2007


RFP reverse


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 7
    Illegal AgeI site found at 119
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

RFP reverse was PCR amplified from mRFP (BBaE1010) using the following primers: 5'-atgcactagtatggcttcctccgaagacgt "Spe mRFP F" 5'-gcattctagattaagcaccggtggagtgac "Xba mRFP R"


Source

RFP reverse was PCR amplified from mRFP (BBaE1010).

References