Difference between revisions of "Part:BBa K731710:Design"

 
 
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<partinfo>BBa_K731710 short</partinfo>
 
<partinfo>BBa_K731710 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The presence of the promoter cannot be tested with screening digestion, as it has more or less the same T7 promoter`s length. Anyway the screening in amplification strains is much more easy: the right colonies will be intensely red.  
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When using and modifying this construct it is important to remember that the two proteins have strong C terminal homology. This should be taken into consideration when designing primers.  
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===Source===
 
===Source===
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The part was built starting from K731700, which is a modified version of a pET21b plasmid where mCherry and A206K Venus were inserted with a 20 bp spacer between the 2 proteins.
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The Ptac promoter has been inserted with a PCR, the sequence of the promoter has been taken from literature,
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K731700  was mutated to replace the T7 terminator with a tac promoter.
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while the original plamid is pET21b
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===References===
 
===References===

Latest revision as of 00:02, 31 August 2012

Platform for terminators analysis under the control of tac promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6865
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 6871
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6865
    Illegal BglII site found at 6011
    Illegal BamHI site found at 6847
    Illegal XhoI site found at 753
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 6865
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 6865
    Plasmid lacks a suffix.
    Illegal XbaI site found at 6880
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 714
    Illegal NgoMIV site found at 1051
    Illegal NgoMIV site found at 4231
    Illegal NgoMIV site found at 4391
    Illegal NgoMIV site found at 5979
    Illegal AgeI site found at 6815
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 2228
    Illegal SapI.rc site found at 3310


Design Notes

When using and modifying this construct it is important to remember that the two proteins have strong C terminal homology. This should be taken into consideration when designing primers.

Source

The part was built starting from K731700, which is a modified version of a pET21b plasmid where mCherry and A206K Venus were inserted with a 20 bp spacer between the 2 proteins. K731700 was mutated to replace the T7 terminator with a tac promoter.

References