Composite

Part:BBa_K4743006:Design

Designed by: Jen Hsien, Liu   Group: iGEM23_PTSH-Taiwan   (2023-09-16)


PnuC-GFP-Pm associated sequence


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1761
    Illegal XbaI site found at 484
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1761
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1761
    Illegal BglII site found at 88
    Illegal BamHI site found at 16
    Illegal XhoI site found at 682
    Illegal XhoI site found at 1408
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1761
    Illegal XbaI site found at 484
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1761
    Illegal XbaI site found at 484
    Illegal AgeI site found at 1774
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

[1]AAGGTACCGCAGGTGGGATCC is the forward primer of Adapter 1

TCACCGGTGCAGGTGGAATTC is the reverse primer of Adapter 2

Also, BamHI is included in adapter 1 and EcoRI is included in Adpater 2. These two site can be used for cloning

[2] We put GFP and Plasma membrane associated sequence intend to check if the transporter pnuC is on the plasma membrane.

Source

Salmonella enterica

References

1. Black, W.B., Aspacio, D., Bever, D. et al. Metabolic engineering of Escherichia coli for optimized biosynthesis of nicotinamide mononucleotide, a noncanonical redox cofactor. Microb Cell Fact 19, 150 (2020). https://doi.org/10.1186/s12934-020-01415-z

2. Popov-Čeleketić, D., Bianchi, F., Ruiz, S. J., Meutiawati, F., & Poolman, B. (2016). A Plasma Membrane Association Module in Yeast Amino Acid Transporters. The Journal of biological chemistry, 291(31), 16024–16037. https://doi.org/10.1074/jbc.M115.706770