Difference between revisions of "Part:BBa K4800003"
Line 11: | Line 11: | ||
<head> | <head> | ||
− | + | <meta charset="UTF-8"> | |
− | + | <meta http-equiv="X-UA-Compatible" content="IE=edge"> | |
− | + | <meta name="viewport" content="width=device-width, initial-scale=1.0"> | |
− | + | <title>Document</title> | |
</head> | </head> | ||
<body> | <body> | ||
− | + | <p style="font-size: 180%; font-weight: bold;">Build: | |
− | + | <p>< img src="https://static.igem.wiki/teams/4800/wiki/parts/ptrc-ycia-sfp-mmcar-yahk-png.png" width="80%" height="80%"></p > | |
− | + | <div align="center"> | |
− | + | <strong>fig1.ptrc</strong> | |
− | + | </div> | |
− | + | <p style="font-size: 160%; font-weight: bold;">1.Experimental Methods | |
− | + | <p>1.1 Construction of PCWJ-YciA-sfp-MmCAR-YahK<br> | |
− | 1. | + | The pCWJ plasmid was extracted from a laboratory conserved strain. In-Fusion cloning was used to ligate the above vector to the Ptrc-YciA-sfp-MmCAR-YahK fragment. |
− | The pCWJ plasmid was extracted from a laboratory conserved strain. In-Fusion cloning was used to ligate the above vector to the Ptrc-YciA-sfp-MmCAR-YahK fragment. | + | <br>1.2 Construction of PRSFDuet-YciA-sfp-MmCAR-YahK<br> |
− | 1. | + | The PRSFDuet plasmid was extracted from laboratory conserved strains. The fragment Ptrc-YciA-sfp-MmCAR-sfp-Yahk was amplified by PCR using PCWJ-YciA-sfp-MmCAR-YahK as a template, and Ptrc-YciA-MmCAR-sfp-Yahk was ligated by In-fusion cloning to the vector PRSFDuet that was linearized using PCR for homologous recombination ligation. |
− | The PRSFDuet plasmid was extracted from laboratory conserved strains. The fragment Ptrc-YciA-sfp-MmCAR-sfp-Yahk was amplified by PCR using PCWJ-YciA-sfp-MmCAR-YahK as a template, and Ptrc-YciA-MmCAR-sfp-Yahk was ligated by In-fusion cloning to the vector PRSFDuet that was linearized using PCR for homologous recombination ligation. | + | </p > |
− | + | ||
<p style="font-size: 180%; font-weight: bold;">Test: | <p style="font-size: 180%; font-weight: bold;">Test: | ||
− | + | </p > | |
− | + | <p style="font-size: 160%; font-weight: bold;">Result | |
− | + | <p>< img src="https://static.igem.wiki/teams/4800/wiki/parts/catalytic-generation-of-1-5-pdo-by-bl21-de3-containing-different-plasmids-png.png" width="80%" height="80%"></p > | |
− | + | <div align="center"> | |
− | + | <strong>fig2.Catalytic generation of 1,5-PDO by BL21(DE3) containing different plasmids</strong> | |
− | <p>By HPLC data, it was seen that BL21 imported into PRSFDuet-YciA-sfp-MmCAR-Yahk catalyzed | + | </div> |
− | + | <p>By HPLC data, it was seen that BL21 imported into PRSFDuet-YciA-sfp-MmCAR-Yahk catalyzed </p > | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</body> | </body> | ||
</html> | </html> | ||
+ | |||
Revision as of 16:29, 6 October 2023
Ptrc-sfp-MmCAR-YahK
(1)Ptrc is a promoter.(2)YahK is a acyl-CoA thioester hydrolase from Haemophilus influenzae.(3) Sfp is a phosphopantetheinyl transferase from Bacilus subtilis.(4) MmCAR is a Carboxylic acid reductase from Mycobacterium marinum.(5) YahK is a NADPH-dependent aldehyde reductase.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1332
Illegal PstI site found at 1767
Illegal PstI site found at 1794
Illegal PstI site found at 3063
Illegal PstI site found at 3090
Illegal PstI site found at 3483
Illegal PstI site found at 3979
Illegal PstI site found at 4623 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1332
Illegal NheI site found at 4891
Illegal PstI site found at 1767
Illegal PstI site found at 1794
Illegal PstI site found at 3063
Illegal PstI site found at 3090
Illegal PstI site found at 3483
Illegal PstI site found at 3979
Illegal PstI site found at 4623 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1332
Illegal BglII site found at 2321
Illegal BglII site found at 2947
Illegal BglII site found at 3223
Illegal BglII site found at 3914
Illegal BamHI site found at 1304
Illegal XhoI site found at 4863 - 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1332
Illegal PstI site found at 1767
Illegal PstI site found at 1794
Illegal PstI site found at 3063
Illegal PstI site found at 3090
Illegal PstI site found at 3483
Illegal PstI site found at 3979
Illegal PstI site found at 4623
Illegal NgoMIV site found at 3591
Illegal NgoMIV site found at 3653
Illegal NgoMIV site found at 3870
Illegal NgoMIV site found at 4317
Illegal AgeI site found at 1497
Illegal AgeI site found at 2508
Illegal AgeI site found at 2805
Illegal AgeI site found at 3690
Illegal AgeI site found at 3699
Illegal AgeI site found at 4374 - 1000COMPATIBLE WITH RFC[1000]