Difference between revisions of "Part:BBa K592200:Design"

 
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__NOTOC__
 
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<partinfo>BBa_K592200 short</partinfo>
 
<partinfo>BBa_K592200 short</partinfo>
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===Design Notes===
 
===Design Notes===
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To make the plasmid usable for Lambda Red recombineering, all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
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To make the plasmid usable for Lambda Red recombineering (Datsenko and Wanner, 2000), all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
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===Source===
 
===Source===
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===References===
 
===References===
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Datsenko, K. A. and B. L. Wanner (2000). "One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products." Proc Natl Acad Sci U S A 97(12): 6640-5.

Revision as of 15:56, 21 September 2011

Low to medium copy BioBrick standard vector


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3116
    Illegal NheI site found at 1403
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3122
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3116
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3116
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3116
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3131
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 1969
    Illegal AgeI site found at 994
    Illegal AgeI site found at 1317
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 289


Design Notes

To make the plasmid usable for Lambda Red recombineering (Datsenko and Wanner, 2000), all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.

Source

Salmonella phage P22 late terminator.

References

Datsenko, K. A. and B. L. Wanner (2000). "One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products." Proc Natl Acad Sci U S A 97(12): 6640-5.