Difference between revisions of "Part:BBa K4806011"
| Line 10: | Line 10: | ||
| − | <p>This basic part contains the coding sequence of the mtTP70C-transit peptide (B2). This part is codon-optimized for <i>Chlamydomonas reinhardtii</i> and was built as part of the CYPurify Collection. In combination with a promotor like PAR like CYP3A4 (<a href=" https://parts.igem.org/Part:BBa_K3002010">BBa_K3002010</a>)<sup>*</sup> and a terminator like tRPL23 (<a href="https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>, this level 0 part leads to expression of your target protein. To detect the target protein a tag like HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>)<sup>*</sup> is recommended.</p> | + | <p>This basic part contains the coding sequence of the mtTP70C-transit peptide to the chloroplast (B2). This part is codon-optimized for <i>Chlamydomonas reinhardtii</i> and was built as part of the CYPurify Collection. In combination with a promotor like PAR like CYP3A4 (<a href=" https://parts.igem.org/Part:BBa_K3002010">BBa_K3002010</a>)<sup>*</sup> and a terminator like tRPL23 (<a href="https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>, this level 0 part leads to expression of your target protein. To detect the target protein a tag like HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>)<sup>*</sup> is recommended.</p> |
<br> | <br> | ||
<h2>Constructs</h2> | <h2>Constructs</h2> | ||
<p> | <p> | ||
| − | <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level-0/ | + | <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level-0/mttp70c-construct.png"> |
<div class="unterschrift"><b>Fig.1 Construct design</b><br> | <div class="unterschrift"><b>Fig.1 Construct design</b><br> | ||
We designed 1 level 2 construct containing the mtTP70C-transit peptide using the modular cloning system (MoClo). | We designed 1 level 2 construct containing the mtTP70C-transit peptide using the modular cloning system (MoClo). | ||
| Line 21: | Line 21: | ||
<p><br></p> | <p><br></p> | ||
<p> | <p> | ||
| − | Here | + | Here is the link to the built construct:<br> |
<ul> | <ul> | ||
| − | <li>1. | + | <li>1. CYPCamC gene for expression in the chloroplast for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806218">BBa_K4806218</a>)</li> |
| − | + | ||
| − | + | ||
| − | + | ||
</ul> | </ul> | ||
</p> | </p> | ||
<p> | <p> | ||
| − | + | This constructs was transformed into <i>Chlamydomonas reinhardtii</i>. Besides the mtTP70C-transit peptide mStop the construct contains the AβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), the CYPCamC coding sequence (<a href=" https://parts.igem.org/Part:BBa_K4806002">BBa_K4806002</a>), the HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>) for detection and the tRPL23-terminator (<a href=" https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>. The resistance cassette for hygromycin is already built in the level 2 vector pMBS810 we are using. The usage of this vector allows the direct assembly of level 0 parts to level 2 constructs, facilitating the cloning time (Niemeyer & Schroda, 2022). | |
</p> | </p> | ||
<h2>Sequence and Features</h2> | <h2>Sequence and Features</h2> | ||
</html> | </html> | ||
| − | <partinfo> | + | <partinfo>BBa_K4806011 SequenceAndFeatures</partinfo> |
| − | <partinfo> | + | <partinfo>BBa_K4806011 parameters</partinfo> |
<html> | <html> | ||
<h2>Results</h2> | <h2>Results</h2> | ||
Revision as of 12:50, 20 September 2023
mtTP70C-transit peptide for Chlamydomonas reinhardtii (Phytobrick)
This basic part contains the coding sequence of the mtTP70C-transit peptide to the chloroplast (B2). This part is codon-optimized for Chlamydomonas reinhardtii and was built as part of the CYPurify Collection. In combination with a promotor like PAR like CYP3A4 (BBa_K3002010)* and a terminator like tRPL23 (BBa_K3002006)*, this level 0 part leads to expression of your target protein. To detect the target protein a tag like HA-tag (BBa_K3002017)* is recommended.
Constructs
Fig.1 Construct design
We designed 1 level 2 construct containing the mtTP70C-transit peptide using the modular cloning system (MoClo).
We designed 1 level 2 construct containing the mtTP70C-transit peptide using the modular cloning system (MoClo).
Here is the link to the built construct:
- 1. CYPCamC gene for expression in the chloroplast for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806218)
This constructs was transformed into Chlamydomonas reinhardtii. Besides the mtTP70C-transit peptide mStop the construct contains the AβSAP(i)-promotor (BBa_K4806013), the CYPCamC coding sequence (BBa_K4806002), the HA-tag (BBa_K3002017) for detection and the tRPL23-terminator (BBa_K3002006)*. The resistance cassette for hygromycin is already built in the level 2 vector pMBS810 we are using. The usage of this vector allows the direct assembly of level 0 parts to level 2 constructs, facilitating the cloning time (Niemeyer & Schroda, 2022).
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2
Illegal PstI site found at 183 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 2
Illegal PstI site found at 183 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 198
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2
Illegal PstI site found at 183 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2
Illegal PstI site found at 183 - 1000COMPATIBLE WITH RFC[1000]