Difference between revisions of "Part:BBa K802005:Design"

 
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<partinfo>BBa_K802005 short</partinfo>
 
<partinfo>BBa_K802005 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The cloning sites of the linker are EcoRI and HindIII (there is no need to digest the linker, the two extremities are sticky ends). Between these sites there are the other 3 iGEM enzyme restriction sites: XbaI, SpeI, PstI.
 
  
  
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<p>The linker was especially designed to contain the iGem restriction sites in the right order. It was obtained by the hybridization of two primers:<br>
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Forward: (5’)aattcgcggccgcttctagagaatactagtagcggccgctgcaga(3’)<br>                                             
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Reverse: (5’)agcttctgcagcggccgctactagtattctctagaagcggccgcg(3’)<br>
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</p>     
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The percentage of the hybridization of the two primers is 91%. The only difference between the two consists in 8 nucleotides: 4 at the 5’end in the case of the Forward primer (which represents the sticky end compatible with a digested HindIII site) and the other 4 at 3’end in the case of the Reverse primer (which represents the sticky end compatible with a digested EcoRI site).
  
 
===Source===
 
===Source===
  
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The linker was obtained by the hybridization of two synthesized oligos.
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The linker was obtained by the hybridization of two synthesized primers. It was designed for cloning in EcoRI and HindIII sites of the polylinker contained by pUC19 (including the shuttle vectors pHT304 or pHT3015 that also have this linker)
  
 
===References===
 
===References===

Latest revision as of 19:15, 26 September 2012

iGEM linker for shuttle vectors BBa_K802003 and BBa_K802004


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 linker was especially designed to contain the iGem restriction sites in the right order. It was obtained by the hybridization of two primers:
Forward: (5’)aattcgcggccgcttctagagaatactagtagcggccgctgcaga(3’)
Reverse: (5’)agcttctgcagcggccgctactagtattctctagaagcggccgcg(3’)

The percentage of the hybridization of the two primers is 91%. The only difference between the two consists in 8 nucleotides: 4 at the 5’end in the case of the Forward primer (which represents the sticky end compatible with a digested HindIII site) and the other 4 at 3’end in the case of the Reverse primer (which represents the sticky end compatible with a digested EcoRI site).

Source

The linker was obtained by the hybridization of two synthesized primers. It was designed for cloning in EcoRI and HindIII sites of the polylinker contained by pUC19 (including the shuttle vectors pHT304 or pHT3015 that also have this linker)

References