Part:BBa_K4585007
pcDNA3.1(+)-3xHA-Gal4-KRAB-NLS linearized vector
The purpose of the linear vector is to perform homologous recombination with the KRAB homologous recombination fragment to obtain the target product pcDNA3.1(+)-3×HA-GAL4-KRAB-NLS plasmid.
pcDNA3.1(+)-3×HA-GAL4-VP64-NLS linearized vector
We based on the sequence of the pcDNA3.1(+) plasmid.The GAL4 sequence, 3×HA sequence were linked into the pcDNA3.1(+) plasmid with the help of homologous recombination experiments. Special primers were designed and then the plasmid was cut into linear sequences of about 5600 bp by PCR.The two ends of this linear sequence can be complementary to the two ends of the KRAB homologous recombination fragment , and the purpose is to conduct homologous recombination with the KRAB homologous recombination fragment to obtain the target product pcDNA3.1 (+) -3×HA-GAL4-KRAB-NLS plasmid.
1 Diagrams
Fig 1. The agarose gel electrophoresis of pcDNA3.1(+)-3×HA-GAL4-KRAB-NLS linearized vector
2 Caution
The two ends of the pcDNA3.1(+)-3×HA-GAL4-KRAB-NLS linearized vector need to be complementary to the two ends of the corresponding recombinant fragments to ensure the success of homologous recombination ligation.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 46
Illegal XbaI site found at 85
Illegal SpeI site found at 4849
Illegal PstI site found at 51
Illegal PstI site found at 1480 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 46
Illegal SpeI site found at 4849
Illegal PstI site found at 51
Illegal PstI site found at 1480
Illegal NotI site found at 72 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 46
Illegal BglII site found at 4612
Illegal BamHI site found at 6048
Illegal XhoI site found at 79
Illegal XhoI site found at 5818 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 46
Illegal XbaI site found at 85
Illegal SpeI site found at 4849
Illegal PstI site found at 51
Illegal PstI site found at 1480 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 46
Illegal XbaI site found at 85
Illegal SpeI site found at 4849
Illegal PstI site found at 51
Illegal PstI site found at 1480
Illegal NgoMIV site found at 590
Illegal NgoMIV site found at 1931
Illegal NgoMIV site found at 2216
Illegal AgeI site found at 143 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5737
Illegal BsaI.rc site found at 3744
Illegal BsaI.rc site found at 5482
Illegal SapI site found at 2661
Illegal SapI.rc site found at 7
Illegal SapI.rc site found at 1780
Illegal SapI.rc site found at 1990
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