Part:BBa_K5207011

pCCW12-SmCPS1-tSSA1

This is an expression cassette consisting of a promoter pCCW12, the SmCPS1 gene, and a terminator tSSA1.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 2397
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1158
  Illegal BamHI site found at 756
  Illegal XhoI site found at 892
  Illegal XhoI site found at 916
  Illegal XhoI site found at 2209
  Illegal XhoI site found at 2419
  Illegal XhoI site found at 2891
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 561
  Illegal NgoMIV site found at 1978
  Illegal NgoMIV site found at 2542
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

We combined each Level 0 junction module, promoter, CDS, terminator, and ending module, and constructed them into vector pYTK095 through BsaI digestion and ligation, and this step was the construction of Level 1.

Figure 1. Snapgene diagrams of pYTK095

After transformation in E. coli DH10B, we selected the single bacterial colony (pYTK095) that does not fluoresce under UV light from the Petri dishes. It was used for colony PCR amplification and validation.

Figure 2. Fluorescent labeling screening of pYTK095

Figure 3. Validation plot of pYTK095 gel electrophoresis