Difference between revisions of "Part:BBa J61032"

Revision as of 21:22, 10 November 2006

Alkaline phosphatase [phoA]

Alkaline phosphatase coding sequence (no rbs, promoter, or terminator present) from Citrobacter. This bacterium's phoA was chosen due simply to the absense of Biobrick restriction sites. There may be some neutral point mutations relative to the stated sequence, but the construct is fully functional. This part exists in plasmid pSB1A2 (pSB1A2-Bca1032), and in J61003 (pBca1020-Bca1032) under the control of a Tet promoter and RBS b0034.

Alkaline phosphatase is used to report gene expression activity. When expressed in the periplasm, PhoA can be assayed by a PNP (para-nitrophenol phosphate) hydrolysis assay--an assay very similar to a standard Miller Assay for LacZ activity. The pre-sequence necessary for periplasmic targetting is encoded within this basic part.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 310
Illegal NgoMIV site found at 757
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_J61032 short</partinfo>
-Alkaline phosphatase coding sequence (no rbs, promoter, or terminator present) from Citrobacter.  This bacterium's phoA was chosen due simply to the absense of Biobrick restriction sites.  There may be some neutral point mutations relative to the stated sequence, but the construct is fully functional.  This part exists in plasmid pSB1A2 (pSB1A2-Bca1032) and in J61003 (pBca1020-Bca1032) under the control of a Tet promoter and RBS b0034.
+Alkaline phosphatase coding sequence (no rbs, promoter, or terminator present) from Citrobacter.  This bacterium's phoA was chosen due simply to the absense of Biobrick restriction sites.  There may be some neutral point mutations relative to the stated sequence, but the construct is fully functional.  This part exists in plasmid pSB1A2 (pSB1A2-Bca1032), and in J61003 (pBca1020-Bca1032) under the control of a Tet promoter and RBS b0034.
-Alkaline phosphatase is used to report gene expression activity.  When expressed in the cytoplasm, PhoA can be assayed by [[Arkin_JCA_PNPAssay | a PNP (para-nitrophenol phosphate) hydrolysis assay]]--an assay very similar to a standard Miller Assay for LacZ activity.
+Alkaline phosphatase is used to report gene expression activity.  When expressed in the periplasm, PhoA can be assayed by [[Arkin_JCA_PNPAssay | a PNP (para-nitrophenol phosphate) hydrolysis assay]]--an assay very similar to a standard Miller Assay for LacZ activity.  The pre-sequence necessary for periplasmic targetting is encoded within this basic part.
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