Difference between revisions of "Part:BBa K5302028"
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This work is derived from pBBR1MCS-INP-mCherry, pUC19-miniZ-Z3C, and it has undergone codon optimization. This composite part combines INP(about 30kda) and Z3C(7kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express Z3C. The plasmid uses lac promotor and has kanamycin resistence. | This work is derived from pBBR1MCS-INP-mCherry, pUC19-miniZ-Z3C, and it has undergone codon optimization. This composite part combines INP(about 30kda) and Z3C(7kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express Z3C. The plasmid uses lac promotor and has kanamycin resistence. | ||
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+ | <hr> | ||
+ | |||
+ | Z3C is derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM). We used pBBR plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C. | ||
+ | |||
+ | <html> | ||
+ | <div style="text-align:center;"> | ||
+ | <img src="https://static.igem.wiki/teams/5302/images/part-registry-z3c-1.png" | ||
+ | width="60%" style="display:block; margin:auto;" alt="Jamboree Program" > | ||
+ | <div style="text-align:center;"> | ||
+ | <caption> | ||
+ | <b>Figure 1. </b> sequence of Z3C and its KD with VEGF | ||
+ | </caption> | ||
+ | </div> | ||
+ | </div> | ||
+ | </html> | ||
+ | |||
+ | <html> | ||
+ | <div style="text-align:center;"> | ||
+ | <img src="https://static.igem.wiki/teams/5302/images/part-registry-z3c-2.png" | ||
+ | width="60%" style="display:block; margin:auto;" alt="Jamboree Program" > | ||
+ | <div style="text-align:center;"> | ||
+ | <caption> | ||
+ | <b>Figure 2. </b> Cartoon representation of the crystal structure of the mini-Z highlighting randomized residues shown as sticks and coloring the helices | ||
+ | </caption> | ||
+ | </div> | ||
+ | </div> | ||
+ | </html> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Latest revision as of 02:03, 2 October 2024
pBBR-INP-Z3C
This work is derived from pBBR1MCS-INP-mCherry, pUC19-miniZ-Z3C, and it has undergone codon optimization. This composite part combines INP(about 30kda) and Z3C(7kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express Z3C. The plasmid uses lac promotor and has kanamycin resistence.
Z3C is derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM). We used pBBR plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4458
Illegal XbaI site found at 3189
Illegal PstI site found at 2016
Illegal PstI site found at 3177 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4458
Illegal PstI site found at 2016
Illegal PstI site found at 3177
Illegal NotI site found at 1057 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4458
Illegal BglII site found at 1803
Illegal BamHI site found at 3195 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4458
Illegal XbaI site found at 3189
Illegal PstI site found at 2016
Illegal PstI site found at 3177 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4458
Illegal XbaI site found at 3189
Illegal PstI site found at 2016
Illegal PstI site found at 3177
Illegal NgoMIV site found at 2467
Illegal NgoMIV site found at 2750
Illegal NgoMIV site found at 3616
Illegal NgoMIV site found at 5123
Illegal AgeI site found at 4963 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1379
Illegal SapI.rc site found at 2316
Illegal SapI.rc site found at 2526