Difference between revisions of "Part:BBa K562009"

 
Line 1: Line 1:
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K562009 short</partinfo>
 
<partinfo>BBa_K562009 short</partinfo>
  
This is a composite part comprising a constitutive promoter, which is the tatABCD promoter from E. coli K-12, driving production of the PduA, PduB, PduT, PduU and PduN proteins from Salmonella enterica serovar Typhimurium LT2 (identical to part BBa_K562008) AND proteins PduJ and PduK. These are all components of the Salmonella bacterial microcompartment (BMC). The PduB, PduU, PduN and PduK gene products all carry C-terminal 6-His affinity/epitope tags. Production of the PduB, PduU, PduN and PduK proteins has been verified by Western immunoblotting (anti-penta-His) and production of PduA, PduB, PduT, PduU, PduN, PduJ and PduK has been confirmed by 35S-Met labelling. The proteins have also been purified from E. coli by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-ABTUNJK in the Sargent Laboratory, Dundee, UK.
+
This is a composite part comprising a constitutive promoter, which is the <i>tatABCD</i> promoter from <i>E. coli</i> K-12, driving production of the PduA, PduB, PduT, PduU and PduN proteins from Salmonella enterica serovar Typhimurium LT2 (identical to part BBa_K562008) AND proteins PduJ and PduK. These are all components of the Salmonella bacterial microcompartment (BMC). The PduB, PduU, PduN and PduK gene products all carry C-terminal 6-His affinity/epitope tags. Production of the PduB, PduU, PduN and PduK proteins has been verified by Western immunoblotting (anti-penta-His) and production of PduA, PduB, PduT, PduU, PduN, PduJ and PduK has been confirmed by 35S-Met labelling. The proteins have also been purified from E. coli by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-ABTUNJK in the Sargent Laboratory, Dundee, UK.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 13:39, 21 September 2011

Salty_PduABTUNJK BMC Components

This is a composite part comprising a constitutive promoter, which is the tatABCD promoter from E. coli K-12, driving production of the PduA, PduB, PduT, PduU and PduN proteins from Salmonella enterica serovar Typhimurium LT2 (identical to part BBa_K562008) AND proteins PduJ and PduK. These are all components of the Salmonella bacterial microcompartment (BMC). The PduB, PduU, PduN and PduK gene products all carry C-terminal 6-His affinity/epitope tags. Production of the PduB, PduU, PduN and PduK proteins has been verified by Western immunoblotting (anti-penta-His) and production of PduA, PduB, PduT, PduU, PduN, PduJ and PduK has been confirmed by 35S-Met labelling. The proteins have also been purified from E. coli by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-ABTUNJK in the Sargent Laboratory, Dundee, UK.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1898
    Illegal BamHI site found at 3058
    Illegal BamHI site found at 3197
    Illegal BamHI site found at 3291
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1441
    Illegal NgoMIV site found at 2685
    Illegal AgeI site found at 1426
    Illegal AgeI site found at 2040
    Illegal AgeI site found at 2952
    Illegal AgeI site found at 2988
  • 1000
    COMPATIBLE WITH RFC[1000]