Difference between revisions of "Part:BBa K2984063"
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This part is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. The backbone is desgined for MoClo level 2 cloning. Together with the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984003">L1a</a>, <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984005">L1b</a> and <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984002"> L1c </a> three different level 1 constructs can be inserted into the level 2 backbone. The backbone contains a transcriptional unit, which we call selection-cassette. This unit consists of the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984008"> Psad </a> promoter, the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984006"> paromomycin resistance paroR </a> and  the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984018"> Rbcs2 </a> terminator. It is flanked by an N-terminal <i>BamHI</i> and a C-terminal <i>XhoI</i> restriction site to allow the exchange of the selection-cassette. Herefore our <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984056"> hygromycin construct </a> or <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984037"> ptxD construct </a>
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This part is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. The backbone is desgined for MoClo level 2 cloning. Together with the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984003">L1a</a>, <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984005">L1b</a> and <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984002"> L1c </a> three different level 1 constructs can be inserted into the level 2 backbone. The backbone contains an antibiotic selection-cassette so this backbone can directly be used for transformations into <i>C. reinhardtii</i>. This unit consists of the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984008"> Psad </a> promoter, the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984006"><i>aphVIII</i> resistance gene for paromomycin resistance</a> and  the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984018"> Rbcs2 </a> terminator. It is flanked by an N-terminal <i>BamHI</i> and a C-terminal <i>XhoI</i> restriction site to allow the exchange of the selection-cassette. Herefore our <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984056"> hygromycin construct </a> or <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984037"> ptxD construct </a>
 
could be inserted instead. The selection-cassette benfits the transformation 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 reduced to one or two using one of our  <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984062"> Level 2 Linker </a> </html>
 
could be inserted instead. The selection-cassette benfits the transformation 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 reduced to one or two using one of our  <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984062"> Level 2 Linker </a> </html>
  

Latest revision as of 14:57, 20 October 2019


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

Level 2 backbone


Usage and Biology

This part is a part of the Chlamy-HUB-Collection. The backbone is desgined for MoClo level 2 cloning. Together with the L1a, L1b and L1c three different level 1 constructs can be inserted into the level 2 backbone. The backbone contains an antibiotic selection-cassette so this backbone can directly be used for transformations into C. reinhardtii. This unit consists of the Psad promoter, the aphVIII resistance gene for paromomycin resistance and the Rbcs2 terminator. It is flanked by an 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 selection-cassette benfits the transformation 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 reduced 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.