Difference between revisions of "Part:BBa K4727002:Design"

 
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===Design Notes===
 
===Design Notes===
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Starting from the construct of Chasteen et al. we deleted the region containing the ORI and the chloramphenicol resistance in order to delete the two PstI cut sites. Following this, a PCR mugaenes was performed on pSBK3 to insert a new MCS downstream of the suffix. Using the cut sites of Alw21I and MluI was possible to clone the M13 genome in the standard backbone
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Starting from the construct of Chasteen et al. we deleted the region containing the ORI and the chloramphenicol resistance in order to delete the two PstI cut sites. Following this, a PCR mugaenes was performed on pSB3K3 to insert a new MCS downstream of the suffix. Using the cut sites of Alw21I and MluI was possible to clone the M13 genome in the standard backbone
  
  

Latest revision as of 20:11, 25 September 2023


M13 helper phage


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 2522
    Illegal suffix found at 2544
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2522
    Illegal NheI site found at 1177
    Illegal SpeI site found at 2545
    Illegal PstI site found at 2559
    Illegal NotI site found at 2528
    Illegal NotI site found at 2552
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2522
    Illegal BglII site found at 2877
    Illegal XhoI site found at 813
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 2522
    Illegal suffix found at 2545
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 2522
    Plasmid lacks a suffix.
    Illegal XbaI site found at 2537
    Illegal SpeI site found at 2545
    Illegal PstI site found at 2559
    Illegal AgeI site found at 1263
    Illegal AgeI site found at 1586
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2291


Design Notes

Starting from the construct of Chasteen et al. we deleted the region containing the ORI and the chloramphenicol resistance in order to delete the two PstI cut sites. Following this, a PCR mugaenes was performed on pSB3K3 to insert a new MCS downstream of the suffix. Using the cut sites of Alw21I and MluI was possible to clone the M13 genome in the standard backbone


Source

Staring from the helper phage designed by Chasteen et al (2006), it was modified by combining it with the standard backbone pSB3K3. The original plasmid has been obtained from the genome of M13 bacteriophage and it carries all the genes coding for the proteins necessary to assemble the viral capsid.

References