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

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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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See also:
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* [https://parts.igem.org/Part:BBa_J18902 pSB1AK3F]
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* [https://parts.igem.org/Part:BBa_J18903 pSB1AT3F]
  
 
===Source===
 
===Source===

Latest revision as of 13:29, 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.

See also:

Source

constructed from pSB1AC3

References