Plasmid

Part:BBa_K1074001

Designed by: Changlong Zhao   Group: iGEM13_USTC_CHINA   (2013-09-09)

PHT43

Usage and Biology

PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. It is based on the strong σA-dependent promoter preceding the groE operon of B. subtilis which has been converted into an efficiently controllable (IPTGinducible)promoter by addition of the lac operator. An efficient Shine-Dalgarno (SD) sequence as well as a multiple cloning site (BamH I, Xba I, Aat II, Sma I) were also inserted. To obtain secretion of recombinant proteins, the coding region for the signal peptide of the amyQ gene encoding an α-amylase was fused to the SD sequence.

Vector Map of pHT43

USTC_China_iGEM13_BBa_K1074001_1.png USTC_China_iGEM13_BBa_K1074001_2.png

Reference

Bacillus subtilis Expression Vectors> MoBiTec GmbH 2012


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1255
    Illegal EcoRI site found at 3507
    Illegal XbaI site found at 2911
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1255
    Illegal EcoRI site found at 3507
    Illegal NheI site found at 1091
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1255
    Illegal EcoRI site found at 3507
    Illegal BglII site found at 2978
    Illegal BglII site found at 6059
    Illegal BamHI site found at 2905
    Illegal XhoI site found at 6063
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1255
    Illegal EcoRI site found at 3507
    Illegal XbaI site found at 2911
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1255
    Illegal EcoRI site found at 3507
    Illegal XbaI site found at 2911
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 6954
    Illegal SapI.rc site found at 4131
    Illegal SapI.rc site found at 8036


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