Plasmid

Part:BBa_K3394007

Designed by: Talia Shaler   Group: iGEM20_BGU-Israel   (2020-10-17)
Revision as of 12:12, 22 October 2020 by Talia sh (Talk | contribs)


Cellulolytic complex [1] with AIDA-I

Our project's goal is to decompose cellulose fibered wet wipes in the sewer systems to prevent the formation of "fatbergs" and clogs. Since cellulose cannot cross the membrane, we engineered E. coli to express cellulolytic complex on its surface through AIDA-1 auto-display system. For more information about AIDA-1 auto-display system, see BBa_K3394006.


Figure 1: decomposition of cellulose fibered wet wipes by engineered E. coli

Our cellulolytic complex is composed of:

1. Cellulase enzyme: We used Endo5a cellulase, which breaks 1,4 beta bonds. In the future, it would help us degrade wet wipes based on cellulose fibers. For more information see BBa_K3394000.

2. CBD-cellulose binding domain. Its purpose is to bind the cellulose polymer thus, enriching the cellulose surface with cellulose degrading bacteria, so the degradation will be more efficient. For more information see BBa_K3394001.

3. His-tag: For more information see BBa_K3394002.

4. GS linker: For more information see BBa_K3394003.

Figure 2: Complete cellulolytic complex expressed on the bacteria's surface by AIDA-1 auto-display system.

We have another design in which the order of the passenger genes is different (BBa_K3394008).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 988
    Illegal EcoRI site found at 1314
    Illegal EcoRI site found at 2117
    Illegal EcoRI site found at 2813
    Illegal EcoRI site found at 6015
    Illegal PstI site found at 2165
    Illegal PstI site found at 2751
    Illegal PstI site found at 5263
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 988
    Illegal EcoRI site found at 1314
    Illegal EcoRI site found at 2117
    Illegal EcoRI site found at 2813
    Illegal EcoRI site found at 6015
    Illegal PstI site found at 2165
    Illegal PstI site found at 2751
    Illegal PstI site found at 5263
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 988
    Illegal EcoRI site found at 1314
    Illegal EcoRI site found at 2117
    Illegal EcoRI site found at 2813
    Illegal EcoRI site found at 6015
    Illegal BamHI site found at 28
    Illegal BamHI site found at 3015
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 988
    Illegal EcoRI site found at 1314
    Illegal EcoRI site found at 2117
    Illegal EcoRI site found at 2813
    Illegal EcoRI site found at 6015
    Illegal PstI site found at 2165
    Illegal PstI site found at 2751
    Illegal PstI site found at 5263
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 988
    Illegal EcoRI site found at 1314
    Illegal EcoRI site found at 2117
    Illegal EcoRI site found at 2813
    Illegal EcoRI site found at 6015
    Illegal PstI site found at 2165
    Illegal PstI site found at 2751
    Illegal PstI site found at 5263
    Illegal NgoMIV site found at 3227
    Illegal NgoMIV site found at 3274
    Illegal NgoMIV site found at 3642
    Illegal NgoMIV site found at 3802
    Illegal NgoMIV site found at 4156
    Illegal AgeI site found at 1289
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5089


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