Difference between revisions of "Part:pSB1A10"

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[[Image:ScreeningPlasmid1.0.PNG|600px|left|thumb|'''Design of Screening Plasmid 1.0:''' We are using the Pbad arabinose-inducible induction system [1] as a tunable input.  GFP is a measure of input and RFP is a measure of output.  A Biobricks cloning site enables easy insertion of any Biobricks part.  RNase E sites create independence between the mRNA stability of the device being screened and the mRNA stability of the fluorescent proteins.  In particular, we suspect mRFP1 contains internal RNaseE cut sites and have added a hairpin 5’ of the coding region to slow degradation by RNase E. [2]
 
[[Image:ScreeningPlasmid1.0.PNG|600px|left|thumb|'''Design of Screening Plasmid 1.0:''' We are using the Pbad arabinose-inducible induction system [1] as a tunable input.  GFP is a measure of input and RFP is a measure of output.  A Biobricks cloning site enables easy insertion of any Biobricks part.  RNase E sites create independence between the mRNA stability of the device being screened and the mRNA stability of the fluorescent proteins.  In particular, we suspect mRFP1 contains internal RNaseE cut sites and have added a hairpin 5’ of the coding region to slow degradation by RNase E. [2]
 
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 09:27, 26 May 2006

Screening Plasmid v1.0

This plasmid can be used to do a rough characterization of the Input/Output curve for a PoPS-based device. Additionally, it can be used for screening of part or device libaries via FACS. More information can be [http://openwetware.org/index.php?title=Endy:Screening_plasmid_1.0 found here.]

Design of Screening Plasmid 1.0: We are using the Pbad arabinose-inducible induction system [1] as a tunable input. GFP is a measure of input and RFP is a measure of output. A Biobricks cloning site enables easy insertion of any Biobricks part. RNase E sites create independence between the mRNA stability of the device being screened and the mRNA stability of the fluorescent proteins. In particular, we suspect mRFP1 contains internal RNaseE cut sites and have added a hairpin 5’ of the coding region to slow degradation by RNase E. [2]


Usage and Biology

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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 5242
    Illegal NheI site found at 4443
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 5248
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5242
    Illegal BglII site found at 89
    Illegal BamHI site found at 4382
    Illegal XhoI site found at 67
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 5242
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 5242
    Plasmid lacks a suffix.
    Illegal XbaI site found at 5257
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal AgeI site found at 674
    Illegal AgeI site found at 786
    Illegal AgeI site found at 4217
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2197
    Illegal BsaI.rc site found at 5118
    Illegal SapI site found at 4199

Functional Parameters

chassis-NA-
copies100-300
mcsBioBrick
originpMB1
resistanceA

References

[1] Khlebnikov et al,Modulation of gene expression from the arabinose-inducible araBAD promoter. J Ind Microbiol Biotechnol. 2002 Jul;29(1):34-7.

[2] Effect of gene location, mRNA secondary structures, and RNase sites on expression of two genes in an engineered operon. Biotechnol Bioeng. 2002 Dec 30;80(7):762-76.