Difference between revisions of "Part:BBa K2804005:Design"
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===Design Notes===
 
===Design Notes===
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It was neccesary to run a PCR to add the basepairs 5'-ccg-3'and 5'-aaaac-3 to the N-terminus and C-terminus of the gBlock in order to allow the enzymatic cleavage by EcoRI and PstI.
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After runing a PCR for adding the basepairs 5'-ccg-3'and 5'-aaaac-3 to the N-terminus and C-terminus of the gBlocks CBD cipA under the control of LacI promoter and sfGFP and then allow the enzymatic cleavage by EcoRI and PstI, a 3A assembly was achieved using psb1c3 as the cloning vector.
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+
  
  
 
===Source===
 
===Source===
  
−
3A assembly
+
Fusion assembly
  
 
===References===
 
===References===
 +
New England Biolabs. Cleavage Close to the End of DNA Fragments (oligonucleotides). https://www.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en

Latest revision as of 16:16, 9 October 2018


CBD cipA fused to sfGFP under the control of LacI promoter


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 NgoMIV site found at 230
    Illegal AgeI site found at 1493
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

After runing a PCR for adding the basepairs 5'-ccg-3'and 5'-aaaac-3 to the N-terminus and C-terminus of the gBlocks CBD cipA under the control of LacI promoter and sfGFP and then allow the enzymatic cleavage by EcoRI and PstI, a 3A assembly was achieved using psb1c3 as the cloning vector.


Source

Fusion assembly

References

New England Biolabs. Cleavage Close to the End of DNA Fragments (oligonucleotides). https://www.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en