Difference between revisions of "Part:BBa K5302011"

 
Line 4: Line 4:
  
 
This work is derived from pBBR1MCS-2, pUC19-OmpA and pET-21b(+)-mCherry, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and mCherry(26.7kda), and we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express mCherry as a fluorescence labeling.The plasmid uses lac promotor and has kanamycin resistence.
 
This work is derived from pBBR1MCS-2, pUC19-OmpA and pET-21b(+)-mCherry, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and mCherry(26.7kda), and we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express mCherry as a fluorescence labeling.The plasmid uses lac promotor and has kanamycin resistence.
 +
 +
<html>
 +
<div style="text-align:center;">
 +
    <img src="https://static.igem.wiki/teams/5302/images/part-registry-pom-1.png"
 +
        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
 +
    <div style="text-align:center;">
 +
        <caption>
 +
            <b>Figure 1. </b> Colony PCR results of pBBR1MCS-OmpA-mCherry
 +
        </caption>
 +
    </div>
 +
</div>
 +
</html>
 +
 +
<html>
 +
<div style="text-align:center;">
 +
    <img src="https://static.igem.wiki/teams/5302/images/part-registry-pom-1.png"
 +
        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
 +
    <div style="text-align:center;">
 +
        <caption>
 +
            <b>Figure 2. </b> structure of OmpA, from genebank
 +
        </caption>
 +
    </div>
 +
</div>
 +
</html>
 +
 +
<html>
 +
<div style="text-align:center;">
 +
    <img src="https://static.igem.wiki/teams/5302/images/part-registry-pom-1.png"
 +
        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
 +
    <div style="text-align:center;">
 +
        <caption>
 +
            <b>Figure 3. </b> SDS-PAGE analysis of pBBR1MCS-OmpA-mCherry expression in Escherichia coli Nissle 1917
 +
        </caption>
 +
    </div>
 +
</div>
 +
</html>
 +
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:24, 1 October 2024


pBBR-OmpA-mCherry

This work is derived from pBBR1MCS-2, pUC19-OmpA and pET-21b(+)-mCherry, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and mCherry(26.7kda), and we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express mCherry as a fluorescence labeling.The plasmid uses lac promotor and has kanamycin resistence.

Jamboree Program
Figure 1. Colony PCR results of pBBR1MCS-OmpA-mCherry

Jamboree Program
Figure 2. structure of OmpA, from genebank

Jamboree Program
Figure 3. SDS-PAGE analysis of pBBR1MCS-OmpA-mCherry expression in Escherichia coli Nissle 1917


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3884
    Illegal EcoRI site found at 3932
    Illegal EcoRI site found at 4205
    Illegal EcoRI site found at 4608
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4193
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3884
    Illegal EcoRI site found at 3932
    Illegal EcoRI site found at 4205
    Illegal EcoRI site found at 4608
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4193
    Illegal NotI site found at 1057
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3884
    Illegal EcoRI site found at 3932
    Illegal EcoRI site found at 4205
    Illegal EcoRI site found at 4608
    Illegal BglII site found at 1803
    Illegal BamHI site found at 3195
    Illegal XhoI site found at 4550
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3884
    Illegal EcoRI site found at 3932
    Illegal EcoRI site found at 4205
    Illegal EcoRI site found at 4608
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4193
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3884
    Illegal EcoRI site found at 3932
    Illegal EcoRI site found at 4205
    Illegal EcoRI site found at 4608
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4193
    Illegal NgoMIV site found at 2467
    Illegal NgoMIV site found at 2750
    Illegal NgoMIV site found at 5266
    Illegal AgeI site found at 4411
    Illegal AgeI site found at 5106
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1379
    Illegal SapI.rc site found at 2316
    Illegal SapI.rc site found at 2526