Difference between revisions of "Part:BBa K1648000"

Latest revision as of 23:07, 24 September 2015

Recombination Template for mamAB Operon

Construct Map for K1648000

Consists of four fragments from magnetosome island of Magnetospirillum gryphiswaldense genome.

Due to potential difficulty in introducing the large magnetosome formation operons into Azotobacter vinelandii, a "template" consists of flanking sequences from mamAB operon is designed for homologous recombination.

Part containing this template (K1648002) 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.

Fig. 1 Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) 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 BamHI site found at 775
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 291
Illegal AgeI site found at 810
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 536

@@ Line 9: / Line 9: @@
 Due to potential difficulty in introducing the large magnetosome formation operons into ''Azotobacter vinelandii'', a "template" consists of flanking sequences from '''mamAB operon''' is designed for '''homologous recombination'''.
-Part containing this template ([https://parts.igem.org/Part:BBa_K1648002 K1648002]) is introduced into ''A. vinelandii'' genome via ' integration''' before full operon sequences to be transformed into ''A. vinelandii''.
+Part containing this template ([https://parts.igem.org/Part:BBa_K1648002 K1648002]) 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.
-[[File:K16480000random.jpg]]
-[fig]Checking of recombinant plasmid using double digestion.
+[[File:K16480000.jpg|400px]]
-L: DNA ladder.
-Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site.
+Fig. 1 Checking of recombinant plasmid using double digestion.
+L: DNA ladder.
+Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site.
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