Plasmid_Backbone

Part:BBa_K2984063:Design

Designed by: Darius Rauch   Group: iGEM19_Humboldt_Berlin   (2019-10-16)
Revision as of 14:13, 20 October 2019 by Chlamylicious (Talk | contribs) (→‎Design Notes)


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 <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Humboldt_Berlin 2019 Chlamy-HUB-Collection</a>. <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 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 <a href="https://2019.igem.org/Team:Humboldt_Berlin/Design#GoldenGate">one-pot reaction</a> via digestion with BbsI generating TGCC and GGGA overhangs 

includes the iGEM RFP-cloning dummy for visual selection of successful insertions.

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

Source

Level 2 Backbone

References