Part:BBa_K4585009

pcDNA3.1(+)-3xHA-Gal4-VP64-Degron-NLS linearized vector

The objective is to perform homologous recombination with the Degron homologous recombination fragment, The synthetic pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS plasmid was used for subsequent experiments.

pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS linearized vector

We based on the sequence of the pcDNA3.1(+)-3×HA-Gal4-VP64-NLS plasmid. Special primers were designed to cut the plasmid into a linear sequence of length of about 6237 bp by PCR. The two ends of this linear sequence can be complementary to the Degron homologous recombination fragment of the homologous Degron fragment for homologous recombination to obtain the target product pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS plasmid.

1 Diagrams

Fig.1 The agarose gel electrophoresis of pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector

2 Caution

The two ends of the pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vectror need to be complementary to the two ends of the corresponding recombinant fragments to ensure the success of homologous recombination ligation.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 25
  Illegal XbaI site found at 64
  Illegal XbaI site found at 6170
  Illegal SpeI site found at 4828
  Illegal PstI site found at 30
  Illegal PstI site found at 1459
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 25
  Illegal SpeI site found at 4828
  Illegal PstI site found at 30
  Illegal PstI site found at 1459
  Illegal NotI site found at 51
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 25
  Illegal BglII site found at 4591
  Illegal BamHI site found at 6027
  Illegal BamHI site found at 6216
  Illegal XhoI site found at 58
  Illegal XhoI site found at 5797
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 25
  Illegal XbaI site found at 64
  Illegal XbaI site found at 6170
  Illegal SpeI site found at 4828
  Illegal PstI site found at 30
  Illegal PstI site found at 1459
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 25
  Illegal XbaI site found at 64
  Illegal XbaI site found at 6170
  Illegal SpeI site found at 4828
  Illegal PstI site found at 30
  Illegal PstI site found at 1459
  Illegal NgoMIV site found at 569
  Illegal NgoMIV site found at 1910
  Illegal NgoMIV site found at 2195
  Illegal AgeI site found at 122
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 5716
  Illegal BsaI.rc site found at 3723
  Illegal BsaI.rc site found at 5461
  Illegal SapI site found at 2640
  Illegal SapI.rc site found at 1759
  Illegal SapI.rc site found at 1969