Difference between revisions of "Part:BBa K5102000:Design"

 
 
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===Design Notes===
 
===Design Notes===
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The construction of pRAM began with a pcDNA3.4-TOPO vector available to us in the lab. Building pRAM required five cloning steps. The first step involved removing the CDS to create an empty vector. Next, the T7 promoter and T7 terminator were introduced. In the third and fourth steps, we removed the HSV TK poly(A) signal, f1 ori, SV40 promoter, and NeoR/KanR. Finally, the last two steps involved introducing point mutations to eliminate BsaI and BgIII restriction sites.
  
  
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===Source===
 
===Source===
  
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.  
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The backbone was cloned from a pcDNA3.4-TOPO vector available to us in the lab.  
  
 
===References===
 
===References===

Latest revision as of 16:58, 1 October 2024


pRAM expression plasmid backbone


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 2989
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 2989
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 2989
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 2989
    Illegal NgoMIV site found at 593
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The construction of pRAM began with a pcDNA3.4-TOPO vector available to us in the lab. Building pRAM required five cloning steps. The first step involved removing the CDS to create an empty vector. Next, the T7 promoter and T7 terminator were introduced. In the third and fourth steps, we removed the HSV TK poly(A) signal, f1 ori, SV40 promoter, and NeoR/KanR. Finally, the last two steps involved introducing point mutations to eliminate BsaI and BgIII restriction sites.


Source

The backbone was cloned from a pcDNA3.4-TOPO vector available to us in the lab.

References