Plasmid_Backbone

Part:BBa_K2984063

Designed by: Darius Rauch   Group: iGEM19_Humboldt_Berlin   (2019-10-16)
Revision as of 14:37, 17 October 2019 by Hermano (Talk | contribs)


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

Level 2 backbone


Usage and Biology

The backbone is desgined for MoClo level 2 cloning. Together with the L1a, L1b and L1c three different level 1 backbone can be inserted into the level 2 backbone. The backbone contains a transcriptional unit, which we call selction-cassette. This unit consists of the Psad promoter, the paromomycin resistance paroR and the Rbcs2 terminator. It is flanked by a N-terminal BamHI and a C-terminal XhoI restriction site to allow the exchange of the selection cassette. Herfore our hygromycin construct or ptxD construct could be inserted instead. The selction cassette benfits the transofrmation rate of Chlamydomonas since it makes co-transformations obsolete. It is not necessary to clone all thre level 1 construct into the backbone. It could be redused to one or two using one of our Level 2 Linker

Sequence and Features


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.


[edit]
Categories
//chassis/eukaryote/plants/other
Parameters
None