Difference between revisions of "Part:BBa K2984063:Design"

 
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===Design Notes===
 
===Design Notes===
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Level 2 Backbone
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This backbone is a part of the Humboldt_Berlin 2019 <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>.
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<a href="https://2019.igem.org/Team:Humboldt_Berlin">The Humboldt_Berlin</a>-team designed a set of Phytobrick backbones optimized for the use in iGEM and <i>C. reinhardtii</i>.
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This backbone is a Phytobrick (MoClo) compatible <b>Level 2 backbone</b>. L2 backbones allow the cloning of multiple L1-gene constructs. L1 constructs can be inserted in a <a href="https://2019.igem.org/Team:Humboldt_Berlin/Design#GoldenGate">one-pot reaction</a> via digestion with <i>Bbs</i>I generating TGCC and GGGA overhangs.
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The backbone includes the iGEM RFP-cloning dummy for visual selection of successful clonings.
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For direct transformation into <i>C. reinhardtii</i>, this backbone includes the <a href="https://parts.igem.org/Part:BBa_K2984055"><i>aphVIII</i></a> selection marker that confers resistance against the antibiotic paromomycin.
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===Source===
 
===Source===
  

Latest revision as of 14:50, 20 October 2019


Phytobrick Level 2 backbone (with C.reinhardtii paromomycin marker)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BamHI site found at 1121
    Illegal XhoI site found at 3012
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal AgeI site found at 189
    Illegal AgeI site found at 301
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


Design Notes

This backbone is a part of the Humboldt_Berlin 2019 Chlamy-HUB-Collection. The Humboldt_Berlin-team designed a set of Phytobrick backbones optimized for the use in iGEM and C. reinhardtii.

This backbone is a Phytobrick (MoClo) compatible Level 2 backbone. L2 backbones allow the cloning of multiple L1-gene constructs. L1 constructs can be inserted in a one-pot reaction via digestion with BbsI generating TGCC and GGGA overhangs.

The backbone includes the iGEM RFP-cloning dummy for visual selection of successful clonings.

For direct transformation into C. reinhardtii, this backbone includes the aphVIII selection marker that confers resistance against the antibiotic paromomycin.

Source

Level 2 Backbone

References