Difference between revisions of "Part:BBa K316008"

 
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<partinfo>BBa_K316008 short</partinfo>
 
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This construct is designed so that the XylE activity is substantially reduced untill such a time when a TEV protease is added to the system and transcribed. TEV protease cleavable linker is positioned between the two proteins. Once the linker is cleaved, XylE is free to tetramerise and assume full activity. GFP is His tagged at the 5' end to facilitate purificaiton for in-vitro assays. Terminator <bbpart>BBa_B0014</bbpart> has been added to comply with Biobrick standards. This particular terminator is stronger and is different from <bbpart>BBa_B0015</bbpart>.
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This part contains <bbpart>BBa_K316007</bbpart> with added double terminator <bbpart>BBa_B0014</bbpart>.  This construct is designed so that the XylE activity is substantially reduced untill such a time when a TEV protease is added to the system and transcribed. TEV protease cleavable linker is positioned between the two proteins. Once the linker is cleaved, XylE is free to tetramerise and assume full activity. GFP is His tagged at the 5' end to facilitate purificaiton for in-vitro assays. Terminator <bbpart>BBa_B0014</bbpart> has been added to comply with Biobrick standards. This particular terminator is stronger and is different from <bbpart>BBa_B0015</bbpart>.
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===Structure and Features===
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[[Image:GFPXylETT.PNG|center|800px]]
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'''Figure I.''' Graphical representation of the GFP-XylE construct with associated Pveg promoter, tags, linkers and double terminator.
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'><big>'''Sequence and Features'''</big></span>
 
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<partinfo>BBa_K316008 parameters</partinfo>
 
<partinfo>BBa_K316008 parameters</partinfo>
 
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For more information about our project please visit our wiki [http://2010.igem.org/Team:Imperial_College_London].

Revision as of 04:19, 26 October 2010

Cleavable GFP-XylE fusion with Pveg promoter and terminator

This part contains BBa_K316007 with added double terminator BBa_B0014. This construct is designed so that the XylE activity is substantially reduced untill such a time when a TEV protease is added to the system and transcribed. TEV protease cleavable linker is positioned between the two proteins. Once the linker is cleaved, XylE is free to tetramerise and assume full activity. GFP is His tagged at the 5' end to facilitate purificaiton for in-vitro assays. Terminator BBa_B0014 has been added to comply with Biobrick standards. This particular terminator is stronger and is different from BBa_B0015.


Structure and Features

GFPXylETT.PNG

Figure I. Graphical representation of the GFP-XylE construct with associated Pveg promoter, tags, linkers and double terminator.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1229
    Illegal NgoMIV site found at 1401
    Illegal AgeI site found at 1752
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 785



For more information about our project please visit our wiki [http://2010.igem.org/Team:Imperial_College_London].