Difference between revisions of "Part:BBa K2336036"
 
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===Usage and Biology===
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<h2>'''Usage and Biology'''</h2>
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<p>PmrC is a promoter used to start the composite. GFP is green fluoresent protein gene.</p>
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<p>PmrC is a promoter used to initiate transcription. GFP is green fluoresent protein gene.</p>
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<p>The aim that we design this part is to get prepared for the material, then PmrA-PmrB(LBT) can be linked to it. Because we need a large amount of this part, we standardize this part independently. Actually, this part is not a functional part. It a basic to get more parts and construct the whole circuit. Figure 1-1 shows the circuit of PmrC-GFP.
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<p>We design this part is for preparation,so pmrA-pmrB(LBT) can link to it. Because we need a large amount of this part, we standardize this part independently. Actually, this part is not a functional part. It the foundation to get more parts and construct the whole circuit. Figure 1 shows the circuit of PmrC-GFP.
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[[File: CGFP.png|left|thumb|400px|Figure 1-1: The circuit of PmrC-GFP.]]</p>
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[[File: CGFP.png|center|thumb|400px|Figure 1: The circuit of PmrC-GFP.]]</p>
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<p></p>
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<h1>'''Characterization'''</h1>
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<h2>'''function'''</h2>
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<p></p>
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<p>When pmrC receive the signal from pmrA-pmrB, it can give the signal to GFP and then green fluorescence can be tested. GFP has restriction enzyme site(XhoI and HindIII), we can cut it and link it with another part, such as sitag.</p>
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<h2>'''experience'''</h2>
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<p>We first get the gene from IDT, then we insert it into pBAD30a. But we discovered that it had SalI restriction enzyme cutting site, so we did point mutation to change it into XhoI. Finally we put the gene into pSB1C3 plasmid. Figure 2 shows the cloning PCR of pmrC-GFP(pSB1C3 plasmid).
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[[File: CGFP1.png|center|thumb|400px|Figure 2: the cloning PCR of PmrC-GFP in pSB1C3 plasmid]]</p>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 20:24, 31 October 2017


PmrC-GFP

This part is used to detect whether the sequence is expressed successfully.


Usage and Biology

PmrC is a promoter used to initiate transcription. GFP is green fluoresent protein gene.

We design this part is for preparation,so pmrA-pmrB(LBT) can link to it. Because we need a large amount of this part, we standardize this part independently. Actually, this part is not a functional part. It the foundation to get more parts and construct the whole circuit. Figure 1 shows the circuit of PmrC-GFP.

Figure 1: The circuit of PmrC-GFP.

function

When pmrC receive the signal from pmrA-pmrB, it can give the signal to GFP and then green fluorescence can be tested. GFP has restriction enzyme site(XhoI and HindIII), we can cut it and link it with another part, such as sitag.

experience

We first get the gene from IDT, then we insert it into pBAD30a. But we discovered that it had SalI restriction enzyme cutting site, so we did point mutation to change it into XhoI. Finally we put the gene into pSB1C3 plasmid. Figure 2 shows the cloning PCR of pmrC-GFP(pSB1C3 plasmid).

Figure 2: the cloning PCR of PmrC-GFP in pSB1C3 plasmid

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 397
    Illegal XhoI site found at 436
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1162
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1085