Difference between revisions of "Part:BBa K2842690"
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This construct is designed to work with [https://parts.igem.org/Part:BBa_K2842680 Intein Monomer 1], it is flanked with the corresponding split intein fragments for protein trans-splicing. An GFP reporter is flanked by the AcelTerL-C intein and the Npu-N intein allowing for polymerisation by protein trans splicing with other proteins flanked by 2 compatible split inteins.
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Like [https://parts.igem.org/Part:BBa_K2842680 Intein Monomer 1] it has SapI cassettes to facilitate the exchange of the sequences that are flanked by inteins. This enables the polymerisation of any protein that can be synthesised.
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[[File:T--UCL--bsaI digests IP RFP GFP.png|400px|thumb|left|
 
[[File:T--UCL--bsaI digests IP RFP GFP.png|400px|thumb|left|

Revision as of 17:46, 17 October 2018


Intein Monomer 2: GFP reporter flanked with orthogonal inteins

Intein Monomer 2
Function Standardised blue-white screening
Use in E. coli cells
Chassis Tested DH5α cells, BL21* cells
Abstraction Hierarchy Composite Device
Related Device BBa_K2842680
RFC standard RFC10,RFC12,RFC21,RFC23
& RFC25 compatible
Backbone pSB1C3
Submitted by [http://2018.igem.org/Team:UCL UCL iGEM 2018]

This construct is designed to work with Intein Monomer 1, it is flanked with the corresponding split intein fragments for protein trans-splicing. An GFP reporter is flanked by the AcelTerL-C intein and the Npu-N intein allowing for polymerisation by protein trans splicing with other proteins flanked by 2 compatible split inteins. Like Intein Monomer 1 it has SapI cassettes to facilitate the exchange of the sequences that are flanked by inteins. This enables the polymerisation of any protein that can be synthesised.

Figure 1: BsaI digestions

(1) BsaI digested Intein Passenger BBa_K2842669
(2) BsaI digested RFP inteins BBa_K2842680
(3) BsaI digested GFP inteins BBa_K2842690
*edited to show relevant bands

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Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1536
    Illegal BsaI.rc site found at 28
    Illegal SapI site found at 1151
    Illegal SapI.rc site found at 422