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 | + | 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> |
| − | could be inserted instead. The | + | 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 redused 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> |
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 23:11, 19 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 a transcriptional unit, which we call selection-cassette. This unit consists of the Psad promoter, the paromomycin resistance paroR 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 redused to one or two using one of our Level 2 Linker
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix. - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix. - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BamHI site found at 1121
Illegal XhoI site found at 3012 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix. - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal AgeI site found at 189
Illegal AgeI site found at 301 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.