Difference between revisions of "Part:BBa K1648003"
 
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[[File:K1648003_Map.png|350px|thumb|right|Construct Map for K1648003]]__NOTOC__
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[[File:K1648003.png|350px|thumb|right|Construct Map for K1648003]]__NOTOC__
 
<partinfo>BBa_K1648003 short</partinfo>
 
<partinfo>BBa_K1648003 short</partinfo>
  
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Consist of four fragments from magnetosome island of ''Magnetospirillum gryphiswaldense'' genome.
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Consist of a template for recombination of <b> mamXY </b>,<b>mamGC </b> and <b> mms </b> operon([https://parts.igem.org/wiki/index.php?title=Part:BBa_K1648001 K1648001]), a promotor & RBS([https://parts.igem.org/Part:BBa_J13002 J13002]) in front of each template and a double terminator([https://parts.igem.org/Part:BBa_B0015 B0015]) at the back of each template.
  
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Due to potential difficulty in introducing the large magnetosome formation operons into ''Azotobacter vinelandii'', a "template" consists of flanking sequences from '''mamAB operons'''except the mamO operon is designed for '''homologous recombination'''.
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This part is designed for <b>homologous recombination</b> to introduce the large magnetosome forming operon into <i>Azotobactor vinelandii</i>.
  
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Part containing this template ([https://parts.igem.org/Part:BBa_K164XXX K1648XXX]) together with another part with mamO operon([https://parts.igem.org/Part:BBa_K164XXX K1648XXX]) is introduced into ''A. vinelandii'' genome via '''random integration''' before full operon sequences to be transformed into ''A. vinelandii''.
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This part is introduced into <i>A. vinelandii</i> genome via <b>random integration</b> before full operon sequences to be transformed into <i>A. vinelandii</i>.
  
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This Part were verified by double digestion (Fig. 1) and sequencing.
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[[File:K1648003gel.jpg|350px]]
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<br>
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Fig 1. Checking of recombinant plasmid using double digestion.
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L: DNA ladder.
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Lane 1-3: Recombination Template for mamXY, mamGC and mms Operons with Promotor and Terminator (BBa_K1648003) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site.
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 04:20, 19 September 2015

Construct Map for K1648003

Recombination Template for mamXY,mamGC and mms Operons with Promotor and Terminator

Consist of a template for recombination of mamXY ,mamGC and mms operon(K1648001), a promotor & RBS(J13002) in front of each template and a double terminator(B0015) at the back of each template.

This part is designed for homologous recombination to introduce the large magnetosome forming operon into Azotobactor vinelandii.

This part is introduced into A. vinelandii genome via random integration before full operon sequences to be transformed into A. vinelandii.

This Part were verified by double digestion (Fig. 1) and sequencing.

K1648003gel.jpg
Fig 1. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for mamXY, mamGC and mms Operons with Promotor and Terminator (BBa_K1648003) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1413
    Illegal BamHI site found at 512
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 683
    Illegal NgoMIV site found at 865
    Illegal NgoMIV site found at 955
    Illegal NgoMIV site found at 1363
    Illegal AgeI site found at 566
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 550
    Illegal BsaI site found at 649
    Illegal BsaI site found at 930