Difference between revisions of "Part:BBa K4806006"

Revision as of 15:37, 20 September 2023

mNeonGreen-tag for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the coding sequence of mNeonGreen (B5). This part is codon-optimized for Chlamydomonas reinhardtii and was built as part of the CYPurify Collection. In combination with a promotor like AβSAP(i) (BBa_K4806013) and a terminator like tRPL23 (BBa_K3002006)^*, this level 0 part leads to detection of your expressed target protein like CYP3A4 (BBa_K4806000).

Constructs

Fig.1 Construct design
We designed 4 level 2 constructs containing mStop using the modular cloning system (MoClo).

Here are the links to the built constructs:

1. The POR gene with mNeonGreen for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806213)
2. CYP9Q3 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806224)
3. CYP3A4 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806204)
4. CYP81A10V7 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806221)

These constructs were transformed into Chlamydomonas reinhardtii. Besides mNeonGreen the constructs contain the CAβSAP(i)-promotor (BBa_K4806013), either the POR (BBa_K4806003), CYP9Q3 (BBa_K4806004), CYP3A4 (BBa_K4806000) or the CYP81A10V7 coding sequence (BBa_K4806005) and the tRPL23-terminator (BBa_K3002006)^*. The resistance cassette for hygromycin or spectinomycin is already built in the level 2 vector pMBS810/pMBS807 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:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 311
Illegal PstI site found at 718
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 311
Illegal PstI site found at 718
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 327
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 311
Illegal PstI site found at 718
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 311
Illegal PstI site found at 718
Illegal AgeI site found at 656
1000
COMPATIBLE WITH RFC[1000]

@@ Line 14: / Line 14: @@
 <h2>Constructs</h2>
 <p>
-   <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level-0/mstop-cons.png">
+   <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level-0/mneongreen-constructs.png">
    <div class="unterschrift"><b>Fig.1 Construct design</b><br>
     We designed 4 level 2 constructs containing mStop using the modular cloning system (MoClo).
@@ Line 23: / Line 23: @@
      Here are the links to the built constructs:<br>
 <ul>
-<li>1. The POR gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806211">BBa_K4806211</a>)</li>
+<li>1. The POR gene with mNeonGreen for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806213">BBa_K4806213</a>)</li>
-<li>2. CYP3A4 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806202">BBa_K4806202</a>)</li>
+<li>2. CYP9Q3 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806224">BBa_K4806224</a>)</li>
-<li>3. CYP9Q3 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806223">BBa_K4806223</a>)</li>
+<li>3. CYP3A4 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806204">BBa_K4806204</a>)</li>
-<li>4. CYP81A10V7 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806220">BBa_K4806220</a>)</li>
+<li>4. CYP81A10V7 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806221">BBa_K4806221</a>)</li>
 </ul>
 </p>
 <p>
-    These constructs were transformed into <i>Chlamydomonas reinhardtii</i>. Besides mStop the constructs contain the CAβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), either the POR (<a href=" https://parts.igem.org/Part:BBa_K4806003">BBa_K4806003</a>), CYP3A3 (<a href=" https://parts.igem.org/Part:BBa_K4806000">BBa_K4806000</a>), CYP9Q3 (<a href=" https://parts.igem.org/Part:BBa_K4806004">BBa_K4806004</a>) or the CYP81A10V7 coding sequence (<a href=" https://parts.igem.org/Part:BBa_K4806005">BBa_K4806005</a>) and the tRPL23-terminator (<a href=" https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>. The resistance cassette for hygromycin or spectinomycin is already built in the level 2 vector pMBS810/pMBS807 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).
+    These constructs were transformed into <i>Chlamydomonas reinhardtii</i>. Besides mNeonGreen the constructs contain the CAβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), either the POR (<a href=" https://parts.igem.org/Part:BBa_K4806003">BBa_K4806003</a>), CYP9Q3 (<a href=" https://parts.igem.org/Part:BBa_K4806004">BBa_K4806004</a>), CYP3A4 (<a href=" https://parts.igem.org/Part:BBa_K4806000">BBa_K4806000</a>) or the CYP81A10V7 coding sequence (<a href=" https://parts.igem.org/Part:BBa_K4806005">BBa_K4806005</a>) and the tRPL23-terminator (<a href=" https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>. The resistance cassette for hygromycin or spectinomycin is already built in the level 2 vector pMBS810/pMBS807 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>
 <h2>Sequence and Features</h2>
 </html>
-<partinfo>BBa_K4806009 SequenceAndFeatures</partinfo>
+<partinfo>BBa_K4806006 SequenceAndFeatures</partinfo>
-<partinfo>BBa_K4806009 parameters</partinfo>
+<partinfo>BBa_K4806006 parameters</partinfo>
 <html>
 <h2>Results</h2>