Composite

Part:BBa_K4268013:Design

Designed by: J. Aubrey, J. Alvarenga, L. Buchanan, J. Reyes, D. Yashinski     Group: iGEM22_SUNY_Oneonta   (2022-09-30)


Tail Tubular Protein B Transcriptional Unit


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2366
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The goal of our team's project, Cyanospectre is to create a ghost phage that could be used by synthetic biologists to make genetic engineering of Cyanobacteria easier. We envision that with modification of this virus, it could be used to immobilize target cyanobacteria, or as a viral vector for the delivery of large circuits into a Cyanobacterial chassis.


We selected a cyanophage, S-TIP37 as the model from which the ghost phage could be built. This phage’s natural host is Synechococcus sp WH8109, a marine Cyanobacteria that has been grown in the lab. We selected this phage based on 1) it has a host that can be grown in the lab, 2) the phage has a relatively small genome making capsid proteins easier to identify, and 3) being a T7-like phage, the structure of its neck region is more simple than that of T4-like phages, making the construction of the ghost phage simpler. We envision that with future modeling, the tail fibers of this ghost phage could be modified to make the virus capable of attaching to and delivering capsid contents to a variety of Salt-water Synechococcus sp strains employed in synthetic biology as chassis, such as Synechococcus sp PCC 11901, UTEX 2973, PCC 7942 or 7002.

Level 0 parts BBa_R0181, BBa_B0034, BBa_K4268005 (Tail-Tubular Protein B), and BBa_B0015 are cloned together to form a Level 1 Transcriptional Unit. According to Type IIS Assembly, there will be 5' to 3' fusion sites between each basic part. The sequence provided contains the fusion sites along with spacer nucleotides that were added for the RBS reading frame.

Source

The source of this sequence is from the S-TIP37 genome (Gene ID 54998412 from the NCBI reference assembly NC_048026.1) (HOT80_gp35 tail tubular protein B [Synechococcus T7-like Phage S-TIP37] - Gene - NCBI, n.d.)

The source of the promoter is a T7 phage Consensus sequence from iGEM's Repository.

The source of the RBS and double terminator are from the group Antiquity on iGEM's Repository.

References

The S-TIP37 genome was found on GenomeNet

U.S. National Library of Medicine. (n.d.). Hot80_gp35 tail tubular protein B [Synechococcus T7-like phage S-tip37] - gene - NCBI. National Center for Biotechnology Information. From https://www.ncbi.nlm.nih.gov/gene/54998412