Difference between revisions of "Part:BBa K4585010"

 
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<partinfo>BBa_K4585010 short</partinfo>
 
<partinfo>BBa_K4585010 short</partinfo>
  
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The purpose of this linear vector is for subsequent homologous recombination synthesis of the pGL4.35-3 HA-9 GAL4UAS-KRAB-Degron-NLS plasmid .
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The purpose of this linear vector is for subsequent homologous recombination synthesis of the pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS plasmid .
  
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<html>
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===Usage and Biology===
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<head>
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4585010 SequenceAndFeatures</partinfo>
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<h2 class="pageContent-main__title">
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            pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS linearized vector
  
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                <p>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.
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                </p>
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            </div>
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            <h2 class="pageContent-main__title pageContent-main__subtitle">
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                1 Pattern Diagram
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                <p style="text-align: center;">
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                    <img width="400px" src="https://static.igem.wiki/teams/4585/wiki/pgl4-35-3-ha-9-gal4uas-krab-degron-nls-linearized-vector/pgl4-35-3-ha-9-gal4uas-krab-degron-nls-linearized-vector.png"></p>
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                <br />
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                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The agarose gel electrophoresis of pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS linearized vector</p>
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            <h2 class="pageContent-main__title pageContent-main__subtitle">
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2 Experiment
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                2.1 Method
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            <div class="pageContent-main__textBox">
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                <p>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
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                </p>
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                <!--put text here, <p>content</p>-->
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                2.2 Results
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                <p style="text-align: center;">
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                    <img width="400px" src="https://static.igem.wiki/teams/4585/wiki/the-model-diagram-of-pgl4-35-3-ha-9-gal4uas-krab-degron-nls111.png"></p>
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                <br />
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                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The model diagram of pGL4.35-3×HA-9×GAL4UAS-KRAB-Degron-NLS</p>
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            </div>
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            <h2 class="pageContent-main__title pageContent-main__subtitle">
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                3.Caution
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            </h2>
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                <p>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.
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                </p>
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            </div>
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</body>
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</html>
  
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===Functional Parameters===
 
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<partinfo>BBa_K4585010 parameters</partinfo>
 
 
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4585009 SequenceAndFeatures</partinfo>

Revision as of 11:54, 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 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