Difference between revisions of "Part:BBa J18901:Design"

 
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<partinfo>BBa_J18901 short</partinfo>
 
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===Design Notes===
 
===Design Notes===
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This plasmid was constructed by PCR and InFusion recombination.  
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This plasmid was constructed by PCR and InFusion recombination.
 +
 
 
1) The pSB1AC3 vector backbone was linearized by PCR, introducing the NgoMIV / AgeI restriction sites
 
1) The pSB1AC3 vector backbone was linearized by PCR, introducing the NgoMIV / AgeI restriction sites
 +
 
2) The P1010 insert was amplified by PCR, introducing the NgoMIV / AgeI restriction sites
 
2) The P1010 insert was amplified by PCR, introducing the NgoMIV / AgeI restriction sites
 +
 
3) The two overlapping PCR products were recombined using the clonetech InFusion kit.
 
3) The two overlapping PCR products were recombined using the clonetech InFusion kit.
  
 
All PCR reactions were performed with a proof-reading polymerase featuring a very low mutation rate. Nevertheless, the vector backbone may contain mutations with respect to pSB1AC3. Only insert and flanks have been verified by sequencing.
 
All PCR reactions were performed with a proof-reading polymerase featuring a very low mutation rate. Nevertheless, the vector backbone may contain mutations with respect to pSB1AC3. Only insert and flanks have been verified by sequencing.
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−
 
  
 
===Source===
 
===Source===

Revision as of 11:26, 11 August 2008

pSB1AC3F construction plasmid for protein fusion BioBricks


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 3043
    Illegal suffix found in sequence at 10
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3043
    Illegal SpeI site found at 11
    Illegal PstI site found at 25
    Illegal NotI site found at 18
    Illegal NotI site found at 3049
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3043
    Illegal XhoI site found at 1044
    Illegal XhoI site found at 1936
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3043
    Illegal suffix found in sequence at 11
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3043
    Illegal suffix found in sequence at 1
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2082


Design Notes

This plasmid was constructed by PCR and InFusion recombination.

1) The pSB1AC3 vector backbone was linearized by PCR, introducing the NgoMIV / AgeI restriction sites

2) The P1010 insert was amplified by PCR, introducing the NgoMIV / AgeI restriction sites

3) The two overlapping PCR products were recombined using the clonetech InFusion kit.

All PCR reactions were performed with a proof-reading polymerase featuring a very low mutation rate. Nevertheless, the vector backbone may contain mutations with respect to pSB1AC3. Only insert and flanks have been verified by sequencing.

Source

constructed from pSB1AC3

References