Part:BBa_K5407000

pLKO.1 puro plasmid

Usage and Biology

pLKO.1 plasmid is a basic backbone plasmid and can be expressed in mammalian cell lines. The plasmid map can be viewed in the figure 1.The backbone size of the plasmid is 7085 bp,and the selectable markers of the plasmid is puromycin. Scientists often use the plasmid tool to inhibit the expression of genes. In our study, we cloned the siRNA sequence targeted to TEAD4 into the basic part pLKO.1 plasmid. And we constructed two composite plasmid named sh-TEAD4-1 and sh-TEAD4-2. After we transfected the plasmid into SW480 cell, the TEAD4 mRNA could be degraded in the cell.

           
                           Figure 1 pLKO.1 map

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1645
  Illegal SpeI site found at 1182
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1645
  Illegal SpeI site found at 1182
  Illegal NotI site found at 1931
  Illegal NotI site found at 2829
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1645
  Illegal BglII site found at 107
  Illegal BglII site found at 148
  Illegal BglII site found at 214
  Illegal BglII site found at 3501
  Illegal BamHI site found at 925
  Illegal XhoI site found at 1444
  Illegal XhoI site found at 1634
  Illegal XhoI site found at 1938
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1645
  Illegal SpeI site found at 1182
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1645
  Illegal SpeI site found at 1182
  Illegal NgoMIV site found at 6296
  Illegal AgeI site found at 1669
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 426
  Illegal BsaI.rc site found at 125
  Illegal BsaI.rc site found at 1340
  Illegal BsaI.rc site found at 3519
  Illegal BsaI.rc site found at 5120
  Illegal SapI site found at 2866
  Illegal SapI site found at 4037