Generator

Part:BBa_K1807002:Design

Designed by: Ivan Gyulev   Group: iGEM15_York   (2015-09-17)
Revision as of 23:54, 18 September 2015 by Isgyulev (Talk | contribs) (→‎Design Notes)

Escherichia coli Polyphosphate Kinase Enzyme


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 396
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1750
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This protein generator device contains the Escherichia coli Polyphosphate Kinase 1 (PPK1) enzyme and has had its illegal Restriction sites removed during assembly.

The part was generated using the following primer pairs:

Fragment 1

Forward: 5' AGCACATACGAGAAAGAGGAGAAATACCCCATGGGTCAGGAAAAGCTATACATCGAAAAA 3'

Reverse: 5' TAGAGGCCGTCGAAcTCCTGATCGGCTTTC 3'

Fragment 2

Forward: 5' GAAAGCCGATCAGGAgTTcGACGGCCTCTA 3'

Reverse: 5' GAGCCTTTCGTTTTATTTGATGCCTGGCCCTTATTCAGGTTGTTCGAGTGATTTGATGT 3'


Fragment 1's For and Fragment 2's Rev Primers contain a point mutation that was designed to remove the illegal EcoRI restriction site.

Source

Escherichia coli BW25113

References