Difference between revisions of "Part:BBa K5407000"

Latest revision as of 11:46, 1 October 2024

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

@@ Line 4: / Line 4: @@
 ===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 7032 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.
+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.
              https://static.igem.wiki/teams/5407/plko.png