Plasmid_Backbone

Part:BBa_K2984063

Designed by: Darius Rauch   Group: iGEM19_Humboldt_Berlin   (2019-10-16)
Revision as of 14:40, 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 constrcuts 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. Herefore 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 three 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