Difference between revisions of "Part:BBa K2984063"

Revision as of 14:37, 17 October 2019

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.

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 <html>
 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 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 <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 a N-terminal BamHI and a C-terminal XhoI restriction site to allow the exchange of the selection cassette. Herfore 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 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  <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984062"> Level 2 Linker </html>
+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  <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>