Part:BBa_K5351001

pSCm-N20

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.
  Illegal NheI site found at 1135
  Illegal NheI site found at 4384
  Illegal NotI site found at 2401
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BamHI site found at 2246
  Illegal BamHI site found at 2357
  Illegal XhoI site found at 2408
  
• 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.
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  

Origin

E. coli; synthesized

Properties

The plasmid pSCm-N20 is a genetic construct containing specific DNA sequences that enable the expression of a particular gene or genes in a host organism. It includes a promoter region for gene expression and possibly other regulatory elements necessary for the function of the inserted gene. It is used as the vector plasmid in our research.

Experimental Approach

Figure 1. Plasmid map of pSCm-N20.

The plasmid pSCm was digested to obtain a linear backbone with a length of 5984 bp.

Figure 2. The gel electrophoresis validation of the linear pSCm-N20 plasmid backbone.

The band shown in Figure 2 indicates that the electrophoresis results are consistent with the expected plasmid size, confirming successful linearization of the plasmid backbone.

Figure 3. The assembly of the pSCm-N20 plasmid with synthesized oligos.

Figure 4. Transformation plate of pSCm-N20.

Figure 5. Sequencing result of pSCm-N20.

Figure 6. PCR validation for integration of PsXI genes.

Figure 7. PCR and colony map of strain xyl-8XI-NFS1.

Figure 8. PCR validation of strain xyl-8XI-ΔISU1.

Figure 9. PCR validation of strain xyl-8XI-nfs1-ΔISU1.

Figure 10. Xylose metabolism analysis using plate assay.

Figure 11. Comparison of xylose metabolism and growth status of modified strains.

Table 1. Ethanol production levels of different strains (48 h).