Difference between revisions of "Part:BBa K3520019"
 
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<partinfo>BBa_K3520019 short</partinfo>
 
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[[File:T--Athens--Level_1_design.png|800px|thumb|center|Figure 1: Type IIS for level 1 Golden Gate assembly with SapI.]]
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==Usage and Biology==
  
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The pMORPHÆ plasmid, consists of the pHimarEm1 vector (BBa_K3520009) containing the transcriptional units of the bcs operon’s genes <i>(A. xylinous)</i>. More specifically, this composite vector was designed in order to permanently insert these genes into the <i>Flavobacterium’s</i> genome, as to produce cellulose. This plasmid was made by connecting the parts  BBa_K3520013, BBa_K3520014, BBa_K3520015, BBa_K3520016, designed by iGEM Athens.
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===Usage and Biology===
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==Plasmid map==
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3520019 SequenceAndFeatures</partinfo>
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[[File:T--Athens--pmorphae_plasmid_map.png|800px|thumb|center|Figure 1: pMorphae complete plasmid map with all TUs.]]
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The present plasmid is used to insert the Bcs operon genes (X54676.1) to a bacterium of the <i>Flavobacterium</i> genus through conjugation using an Escherichia coli strain for conjugation. Each gene has its own transcriptional unit that will allow their constitutive expression. These genes will all intergrated into the genome of the target <i>Flavobacterium</i> with the help of the Mariner transposon and then the Cellulose is produced by the recombinant bacteria.
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==Type IIS assembly design==
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[[File:T--Athens--Level_1_design.png|800px|thumb|center|Figure 2: Type IIS for level 1 Golden Gate assembly with SapI.]]
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==Gene circuitry==
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[[File:T--Athens--bc_gene_circuit.png|800px|thumb|center|Figure 3: The interactions regulating the production of bacterial cellulose under the bcs operon.]]
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==Athens 2020==
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<br>
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The current part is designed by iGEM Athens 2020 team during the project MORPHÆ. In this project, Flavobacteria were used to produce a non-cellular structurally coloured biomaterial which would require the secretion of a biomolecule that Flavobacteria do not normally secrete. Our hypothesis is that the formed matrix will have a structure similar to that of the biofilm and thus, it will provide the material with macroscopically the same colouration properties as the biofilm.
  
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===Functional Parameters===
 
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<partinfo>BBa_K3520019 parameters</partinfo>
 
 
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3520019 SequenceAndFeatures</partinfo>

Latest revision as of 03:43, 28 October 2020


pHimarEm1+TUs of BcsA,B,C,D

Usage and Biology

The pMORPHÆ plasmid, consists of the pHimarEm1 vector (BBa_K3520009) containing the transcriptional units of the bcs operon’s genes (A. xylinous). More specifically, this composite vector was designed in order to permanently insert these genes into the Flavobacterium’s genome, as to produce cellulose. This plasmid was made by connecting the parts BBa_K3520013, BBa_K3520014, BBa_K3520015, BBa_K3520016, designed by iGEM Athens.

Plasmid map

Figure 1: pMorphae complete plasmid map with all TUs.

The present plasmid is used to insert the Bcs operon genes (X54676.1) to a bacterium of the Flavobacterium genus through conjugation using an Escherichia coli strain for conjugation. Each gene has its own transcriptional unit that will allow their constitutive expression. These genes will all intergrated into the genome of the target Flavobacterium with the help of the Mariner transposon and then the Cellulose is produced by the recombinant bacteria.

Type IIS assembly design

Figure 2: Type IIS for level 1 Golden Gate assembly with SapI.

Gene circuitry

Figure 3: The interactions regulating the production of bacterial cellulose under the bcs operon.

Athens 2020


The current part is designed by iGEM Athens 2020 team during the project MORPHÆ. In this project, Flavobacteria were used to produce a non-cellular structurally coloured biomaterial which would require the secretion of a biomolecule that Flavobacteria do not normally secrete. Our hypothesis is that the formed matrix will have a structure similar to that of the biofilm and thus, it will provide the material with macroscopically the same colouration properties as the biofilm.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 16429
    Illegal suffix found in sequence at 9767
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 16429
    Illegal NheI site found at 10096
    Illegal SpeI site found at 9768
    Illegal PstI site found at 9782
    Illegal NotI site found at 9775
    Illegal NotI site found at 16435
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 16429
    Illegal BglII site found at 10418
    Illegal BglII site found at 12957
    Illegal BglII site found at 13616
    Illegal XhoI site found at 10053
    Illegal XhoI site found at 13254
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 16429
    Illegal suffix found in sequence at 9768
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 16429
    Illegal XbaI site found at 16444
    Illegal SpeI site found at 9768
    Illegal PstI site found at 9782
    Illegal NgoMIV site found at 11082
    Illegal AgeI site found at 12649
  • 1000
    COMPATIBLE WITH RFC[1000]