Part:BBa_K2984063:Design

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===