Difference between revisions of "Part:pSB1A10"

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.]

Schematic 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

-- Please enter your experience with this part here --

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-
copies	100-300
mcs	BioBrick
origin	pMB1
resistance	A

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.

@@ Line 4: / Line 4: @@
 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.]
-[[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|'''Schematic 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]
 ]]
 <br style="clear:both" />