Difference between revisions of "Part:BBa K243033"

 
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<partinfo>BBa_K243033 short</partinfo>
 
<partinfo>BBa_K243033 short</partinfo>
  
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This is our expression vector. The plasmid has the restriction sites XbaI NgoMIV_part_AgeI_SpeI_NotI_PstI
 
  
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This is our main E. coli protein expression vector. To insert DNA fragments, the plasmid possesses the restriction sites XbaI NgoMIV and AgeI_SpeI_NotI_PstI. The IPTG-inducable plasmid vector contains the pBR322 origin of replication and a Shine-Dalgarno-Sequence.
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[[Image:Freiburg09 pEx.JPG]]
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===Usage and Biology===
 
===Usage and Biology===
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We used this vector containing an ampicillin resistance gene for the expression of single constructs such as the His-FluA-Split-Fok_i (BBa_K243010)construct. Simultaneous expression of the other Fok construct, His-Dig-Split-Fok_a (BBa_K243036) could be assured using the vector pJS419 since it confers chloramphenicol resistance to the host cell. This enables a positive selection of bacterial strains such as XL1-blue (Stratagene) cotransformed with both plasmids.
  
 
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>'''Sequence and Features'''</span>
 
<partinfo>BBa_K243033 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K243033 SequenceAndFeatures</partinfo>
  

Latest revision as of 19:36, 23 October 2009

pEX


This is our main E. coli protein expression vector. To insert DNA fragments, the plasmid possesses the restriction sites XbaI NgoMIV and AgeI_SpeI_NotI_PstI. The IPTG-inducable plasmid vector contains the pBR322 origin of replication and a Shine-Dalgarno-Sequence.


Freiburg09 pEx.JPG


Usage and Biology

We used this vector containing an ampicillin resistance gene for the expression of single constructs such as the His-FluA-Split-Fok_i (BBa_K243010)construct. Simultaneous expression of the other Fok construct, His-Dig-Split-Fok_a (BBa_K243036) could be assured using the vector pJS419 since it confers chloramphenicol resistance to the host cell. This enables a positive selection of bacterial strains such as XL1-blue (Stratagene) cotransformed with both plasmids.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal suffix found in sequence at 2734
    Illegal XbaI site found at 1523
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 2735
    Illegal PstI site found at 2749
    Illegal NotI site found at 2742
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1808
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal suffix found in sequence at 2735
    Illegal XbaI site found at 1523
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal suffix found in sequence at 2725
    Illegal XbaI site found at 1523
    Illegal NgoMIV site found at 1531
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3800
    Illegal SapI.rc site found at 4799