Plasmid_Backbone

Part:BBa_K2984063:Design

Designed by: Darius Rauch   Group: iGEM19_Humboldt_Berlin   (2019-10-16)
Revision as of 14:50, 20 October 2019 by Chlamylicious (Talk | contribs) (→‎Design Notes)

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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