Difference between revisions of "Part:BBa K1433019"

 
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<partinfo>BBa_K1433019 short</partinfo>
 
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<P>Lambda red is a Lambda-phage-derived recombination system which contains three proteins: Exo, Beta and Gam. It can recombine dsDNA/ssDNA into different kinds of DNA molecules such as chromosome, bacteria artificial chromosome (BAC) or even multicopy plasmids, as long as each side of the donor dsDNA/ssDNA is flanked by 36-50bp homologous arms. The homologous arms also determine the accurate sites of recombination, which ensures that the donor fragment can be inserted into any sites by choosing and adding homologous arms using PCR.</P>
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<P>This part is designed to test lambda red recombination.</P>
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<p><b><big>Composition:</big></b><br />
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<ol><li>Promoter:  [https://parts.igem.org/Part:BBa_J23110 BBa_J23110], a middle-ground Bacterial constitutive promoter.</li>
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<li>Homologous arms:  homologous arms A and B.</li>
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<li>Terminator: [https://parts.igem.org/Part:BBa_B0015 BBa_B0015], a strong double terminator. </li>
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<li>RBS: [https://parts.igem.org/Part:BBa_B0034 BBa_B0034], a strong RBS.</li>
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<li>GFP: [https://parts.igem.org/Part:BBa_E0040 BBa_E0040], green fluorescent protein derived from jellyfish Aequeora Victoria wild-type GFP.</li></ol></p>
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[[File:singleoverview.jpg]]
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<p>In this circuit, two terminators suppress the expression of GFP until recombination occurs between homologous arms A and B. The inserted sequence replaces these two terminators and enables the expression of the downstream GFP. The green color of the strain can be easily observed.</p>
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[[File:lambda red 重组小动画.gif]]
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Revision as of 18:24, 16 October 2014

P-A-T-T-B-B0034-GFP-T

Lambda red is a Lambda-phage-derived recombination system which contains three proteins: Exo, Beta and Gam. It can recombine dsDNA/ssDNA into different kinds of DNA molecules such as chromosome, bacteria artificial chromosome (BAC) or even multicopy plasmids, as long as each side of the donor dsDNA/ssDNA is flanked by 36-50bp homologous arms. The homologous arms also determine the accurate sites of recombination, which ensures that the donor fragment can be inserted into any sites by choosing and adding homologous arms using PCR.

This part is designed to test lambda red recombination.

Composition:

  1. Promoter: BBa_J23110, a middle-ground Bacterial constitutive promoter.
  2. Homologous arms: homologous arms A and B.
  3. Terminator: BBa_B0015, a strong double terminator.
  4. RBS: BBa_B0034, a strong RBS.
  5. GFP: BBa_E0040, green fluorescent protein derived from jellyfish Aequeora Victoria wild-type GFP.

Singleoverview.jpg

In this circuit, two terminators suppress the expression of GFP until recombination occurs between homologous arms A and B. The inserted sequence replaces these two terminators and enables the expression of the downstream GFP. The green color of the strain can be easily observed.

File:Lambda red 重组小动画.gif


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1048