Difference between revisions of "Part:BBa K4585010"
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<span class='h3bb'>Sequence and Features</span>
 
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Revision as of 11:55, 9 October 2023


pGL4.35-3xHA-9xGAL4UAS-KRAB-Degron-NLS linearized vector

The purpose of this linear vector is for subsequent homologous recombination synthesis of the pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS plasmid .

pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS linearized vector

We based on the sequence of the pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS plasmid. Special primers were designed to cut the plasmid into a linear sequence of length of about 5338bp 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 pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS plasmid.

1 Pattern Diagram


Fig.1 The agarose gel electrophoresis of pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS linearized vector

2 Experiment

2.1 Method

Based on the pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS plasmid, we designed the new pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS linearized vectror

2.2 Results


Fig.1 The model diagram of pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS

3.Caution

The two ends of the pGL4.35-3×HA-9×GAL4UAS-KRAB-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 4433
    Illegal XbaI site found at 161
    Illegal XbaI site found at 1943
    Illegal XbaI site found at 4270
    Illegal XbaI site found at 4505
    Illegal SpeI site found at 4029
    Illegal PstI site found at 3118
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4433
    Illegal NheI site found at 4461
    Illegal SpeI site found at 4029
    Illegal PstI site found at 3118
    Illegal NotI site found at 3097
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4433
    Illegal BglII site found at 4508
    Illegal BamHI site found at 423
    Illegal BamHI site found at 5100
    Illegal BamHI site found at 5325
    Illegal XhoI site found at 4313
    Illegal XhoI site found at 4870
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4433
    Illegal XbaI site found at 161
    Illegal XbaI site found at 1943
    Illegal XbaI site found at 4270
    Illegal XbaI site found at 4505
    Illegal SpeI site found at 4029
    Illegal PstI site found at 3118
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4433
    Illegal XbaI site found at 161
    Illegal XbaI site found at 1943
    Illegal XbaI site found at 4270
    Illegal XbaI site found at 4505
    Illegal SpeI site found at 4029
    Illegal PstI site found at 3118
    Illegal NgoMIV site found at 65
    Illegal NgoMIV site found at 176
    Illegal NgoMIV site found at 1654
    Illegal NgoMIV site found at 1671
    Illegal NgoMIV site found at 1803
    Illegal NgoMIV site found at 1894
    Illegal NgoMIV site found at 2146
    Illegal AgeI site found at 1949
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4789
    Illegal SapI site found at 444
    Illegal SapI site found at 2155
    Illegal SapI.rc site found at 5283