Coding

Part:BBa_K4268000:Design

Designed by: J. Aubrey, J. Alvarenga, L. Buchanan, J. Reyes, D. Yashinski   Group: iGEM22_SUNY_Oneonta   (2022-07-15)
Revision as of 11:19, 23 September 2022 by Jazmine124 (Talk | contribs) (→‎Design Notes)


S-TIP37 Head-Tail Connector


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 346
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To determine which sequences would be used to code for the shell of the phage, a series of comparisons were made. SUNY_Oneonta iGEM had identified T7 and T4-like phages. The team chose to utilize T7-like phages due to their smaller genome compared to T4-like phages. The smaller genome would allow greater efficiency and ease in cloning the chosen species' genes.

Synechococcus sp PCC 11901, was decided as the desired cyanobacterial species that SUNY_Oneonta iGEM would like to focus on? S-TIP 37 is the infecting cyanophage of this cyanobacterial species. However, the genome has not undergone full sequencing and many of its genes remain hypothetical proteins. Thus, comparison to Syn-5, another T7-like phage that has been fully sequenced, was needed. Both S-TIP 37 and Syn-5 are structurally and genetically similar allowing the team to distinguish the genes needed for producing the phage shell.


Insert poster table comparison


Upon the isolation of the Head-Tail Connector sequence, the following edits were made:

  • Restriction sites were removed/altered to ensure RCF10 and RCF1000 compatibility
  • To complete Level 0 Golden Gate Assembly, the sequence contains SapI sites at the ends

Source

The source of this sequence is from the S-TIP37 genome (GenBanK: NC_048026.1)

References

The S-TIP37 genome was found on GenomeNet