Difference between revisions of "Part:BBa K4585009"
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The objective is to perform homologous recombination with the Degron  homologous recombination fragment , The synthetic pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS plasmid was used for subsequent experiments.
 
The objective is to perform homologous recombination with the Degron  homologous recombination fragment , The synthetic pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS plasmid was used for subsequent experiments.
  
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4585009 SequenceAndFeatures</partinfo>
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            pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector
  
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===Functional Parameters===
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<partinfo>BBa_K4585009 parameters</partinfo>
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                <p>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 6237bp 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.
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            <h2 class="pageContent-main__title pageContent-main__subtitle">
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                1 Pattern Diagram
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                    <img width="400px" src="https://static.igem.wiki/teams/4585/wiki/pcdna3-1-3-ha-gal4-vp64-degron-nls-linearized-vector/pcdna3-1-3-ha-gal4-vp64-degron-nls-linearized-vector.png"></p>
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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 pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector</p>
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2 Experiment
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                2.1 Method
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                <p>Based on the designed pcDNA3.1 (+) -3 HA-Gal 4-VP64-NLS plasmid , Design of the new pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS linearized vectror
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                2.2 Results
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                    <img width="400px" src="https://static.igem.wiki/teams/4585/wiki/the-model-diagram-of-pcdna3-1-3-ha-gal4-vp64-degron-nls111.png"></p>
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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 pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS</p>
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                3.Caution
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                <p>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.
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Revision as of 11:32, 9 October 2023


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-Gal 4-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 6237bp 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 Pattern Diagram


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

2 Experiment

2.1 Method

Based on the designed pcDNA3.1 (+) -3 HA-Gal 4-VP64-NLS plasmid , Design of the new pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS linearized vectror

2.2 Results


Fig.1 The model diagram of pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS

3.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.