Difference between revisions of "Part:BBa K4012017"
 
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[[Image:CDS Pc4CL.jpeg|thumbnail|750px|center|'''Figure 2:'''  
 
[[Image:CDS Pc4CL.jpeg|thumbnail|750px|center|'''Figure 2:'''  
 
[https://parts.igem.org/Part:BBa_K4012017] A: the construction strategy; B: results of colony PCR; C: results of sequencing analysis of coding sequence Pc4CL]]
 
[https://parts.igem.org/Part:BBa_K4012017] A: the construction strategy; B: results of colony PCR; C: results of sequencing analysis of coding sequence Pc4CL]]
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The construction schematic of GbCHS sequence demonstrated as Fig .2. The initiation of the GbCHS sequence is done by promoter pTEF1, with termination done by tADH1. The sequence ConL2 and ConR3 are connector sequences within the Level 1 plasmid assembly. Furthermore, typr9-KVF and type9-VR are imposed to enact selection through colonies PCR, results shown in Fig.2 The band length 2500bp match with expectations. The sequence analysis results show no significant mutations or deletions, representing the success of the assembly.
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The construction schematic of Pc4CL sequence demonstrated in Fig.5. The initiation of the Pc4CL sequence is done by promoter pPGK1, with termination done by tCYC1. The sequence ConL1 and ConR2 are connector sequences within the Level 1 plasmid assembly. Furthermore, type9-KVF and type9-VR are imposed to enact selection through colonies PCR, results shown in Fig.5 B The band length 3004 bp match with expectations. The sequence analysis results show no significant mutations or deletions, representing the success of the assembly.
  
 
===Pc4CL in Level2 plasmid assembly===
 
===Pc4CL in Level2 plasmid assembly===
 
[[Image:yeast PCR.jpg|thumbnail|750px|center|'''Figure 3:'''  
 
[[Image:yeast PCR.jpg|thumbnail|750px|center|'''Figure 3:'''  
 
[https://parts.igem.org/Part:BBa_K4012017] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway]]
 
[https://parts.igem.org/Part:BBa_K4012017] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway]]
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GbCHS is successfully inserted into the vector type9 C-mTurquoise-Ura.
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Pc4CL is successfully inserted into the vector type9 C-mTurquoise-Ura.

Latest revision as of 13:19, 21 October 2021


Pc4CL

Produces CoA thioesters of a variety of hydroxy- and methoxy-substituted cinnamic acids, which are used to synthesize several phenylpropanoid-derived compounds, including anthocyanins, flavonoids, isoflavonoids, coumarins, lignin, suberin and wall-bound phenolics.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Pc4CL in Level1 plasmid assembly

Figure 2: [1] A: the construction strategy; B: results of colony PCR; C: results of sequencing analysis of coding sequence Pc4CL

The construction schematic of Pc4CL sequence demonstrated in Fig.5. The initiation of the Pc4CL sequence is done by promoter pPGK1, with termination done by tCYC1. The sequence ConL1 and ConR2 are connector sequences within the Level 1 plasmid assembly. Furthermore, type9-KVF and type9-VR are imposed to enact selection through colonies PCR, results shown in Fig.5 B The band length 3004 bp match with expectations. The sequence analysis results show no significant mutations or deletions, representing the success of the assembly.

Pc4CL in Level2 plasmid assembly

Figure 3: [2] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway

Pc4CL is successfully inserted into the vector type9 C-mTurquoise-Ura.