Difference between revisions of "Part:BBa K2350009"

Revision as of 16:54, 22 October 2017

Pigment double-crossover homologous gene recombination bacbone (pPIGBACK)

The vector, pPIGBACK, is used to transform into S. elongatus PCC 7942 with the inserted pigment gene in our project. pPIGBACK contains 5’- and 3’-ends of the neutral site II (NSII), replication origin of pBR322, ampicillin resistance gene, and double terminator BBa_B0015. The strategy we chose to construct the vector is to fuse B0015 and AmpR together first. Secondly, we fused 5’- and 3’-ends of the neutral site II (NSII) with PBR322 replication origin (ORI) together. At last, we ligated two parts together. The aim of constructing this vector is to finish double-crossover homologous gene recombination in S. elongatus PCC 7942. To insert BBa_J04450 with EcoR1 and Pst1 cutting sites, we use site-directed mutagenesis to remove all Pst1 cutting sites in pPIGBACK.

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal prefix found at 1
Illegal suffix found at 3880
12
INCOMPATIBLE WITH RFC[12]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 1
Illegal SpeI site found at 3881
Illegal PstI site found at 3895
Illegal NotI site found at 7
Illegal NotI site found at 1321
Illegal NotI site found at 3888
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 1
Illegal BglII site found at 1883
Illegal BamHI site found at 2180
Illegal XhoI site found at 1118
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found at 1
Illegal suffix found at 3881
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found at 1
Plasmid lacks a suffix.
Illegal XbaI site found at 16
Illegal SpeI site found at 3881
Illegal PstI site found at 3895
Illegal NgoMIV site found at 2101
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 972
Illegal BsaI site found at 1783

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 __NOTOC__
 <partinfo>BBa_K2350009 short</partinfo>
 The vector, pPIGBACK, is used to transform into S. elongatus PCC 7942 with the inserted pigment gene in our project. pPIGBACK contains 5’- and 3’-ends of the neutral site II (NSII), replication origin of pBR322, ampicillin resistance gene, and double terminator BBa_B0015.