Reporter

Part:BBa_J61032

Designed by: John Anderson   Group: Arkin Lab   (2006-11-10)

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.

The reporter gene phoA encoding 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]


[edit]
Categories
//cds/enzyme
//function/reporter/color
Parameters
None