Difference between revisions of "Part:BBa K731700"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K731700 short</partinfo>
 
<partinfo>BBa_K731700 short</partinfo>
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We used this part to test terminators: the two subsequent proteins allow to quantify the termination efficiency as the ratio between Venus's and Cherry's fluorescence.
 
We used this part to test terminators: the two subsequent proteins allow to quantify the termination efficiency as the ratio between Venus's and Cherry's fluorescence.
 
The vector is pET21b, that contains lacI, t7 promoter and ampicillin resistance gene.
 
The vector is pET21b, that contains lacI, t7 promoter and ampicillin resistance gene.
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The two fluorescent proteins have partially overlapping emission spectra, thus the fluorimetric measures should be taken with these parameters, to avoid interference between the two:
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The two fluorescent proteins have partially overlapping emission spectra, thus, to avoid unwanted interferences, the fluorimetric measures should be taken with these parameters:
 
Venus excitation: 485nm, Venus emission: 528nm
 
Venus excitation: 485nm, Venus emission: 528nm
 
Cherry excitation: 528nm, Cherry emission: 615nm
 
Cherry excitation: 528nm, Cherry emission: 615nm

Revision as of 20:00, 23 August 2012

Platform for terminators analysis under the control of T7 promoter

We used this part to test terminators: the two subsequent proteins allow to quantify the termination efficiency as the ratio between Venus's and Cherry's fluorescence. The vector is pET21b, that contains lacI, t7 promoter and ampicillin resistance gene. The two fluorescent proteins have partially overlapping emission spectra, thus, to avoid unwanted interferences, the fluorimetric measures should be taken with these parameters: Venus excitation: 485nm, Venus emission: 528nm Cherry excitation: 528nm, Cherry emission: 615nm We inserted the prefix-suffix linker between the two proteins, to allow terminator characterization.

Sequence and Features


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 6850
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 6856
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6850
    Illegal BglII site found at 6011
    Illegal BamHI site found at 6832
    Illegal XhoI site found at 753
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 6850
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 6850
    Plasmid lacks a suffix.
    Illegal XbaI site found at 6865
    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 6800
  • 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