Rationale:

To annotate genes 18, 19, and 20 of the phage NapoleonB.

Procedures:

1. DNA Master was opened with the fasta file for NapoleonB being selected.
2. NapoleonB’s sequence was auto-annotated.
3. The ORF screen was opened and the RBS values were opened as well.
4. Gene 18 was then annotated.
5. Steps 2-4 were repeated for genes 19 and 20 respectively.

Results:

Gene 18:

Gene 19:

Gene 20:

Gene 18:

SSC:14865 – 16796, CP:Yes, SCS:Both, ST:SS, BLAST-Start:Aligns with KeaneyLin gp NCBI BLAST q1:s1 0.99 0, Aligns with KeaneyLin gp18 PhagesDB BLAST q1:s1 0.99 0, Gap:95bp gap, LO:No, RBS:Kibbler7 and Karlin Medium 1.803 -5.081 No, F:minor tail protein, SIF-BLAST:Minor Tail Protein Supported by NCBI BLAST KeaneyLin gpNA AXH44156.1 0.99 0, Minor Tail Protein Supported by PhagesDB BLAST KeaneyLin gp18 0.99 0, GDSL-like Lipase/Acylhydrolase supported by a CD found from the pfam database gp pfam13472 NA 1.01E-11, , SIF-HHPred:NKF, SIF-Syn:minor tail protein, upstream gene is major tail protein, downstream gene is NKF, just like in phage Nason

Gene 19:

SSC:16816 – 17196, CP:Yes, SCS:BothGL, ST:SS, BLAST-Start:Aligns with KeaneyLin gp NCBI BLAST q1:s1 0.99 3E-86, Aligns with KeaneyLin gp19 PhagesDB BLAST q1:s1 0.98 6E-69, Gap:19bp gap, LO:Yes, RBS:Kibbler7 and Karlin Medium 2.381 -3.86 Yes, F:minor tail protein, SIF-BLAST:Minor Tail Protein Supported by NCBI BLAST KeaneyLin gpNA AXH44157.1 0.99 3E-86, Minor Tail Protein Supported by PhagesDB BLAST KeaneyLin gp19 0.98 6E-69, putative tail-component supported by a CD found from the pfam database gp pfam04883 NA 0.00924, , SIF-HHPred:NKF, SIF-Syn:NKF, upstream gene is minor tail protein, downstream gene is NKF, just like in phage Nason

Gene 20:

SSC:17193 – 21671, CP:Yes, SCS:Both, ST:SS, BLAST-Start:Aligns with Arcadia gp NCBI BLAST q1:s1 0.99 0, Aligns with Nason gp22 PhagesDB BLAST q1:s1 0.99 0, Gap:4bp overlap, LO:NA, RBS:Kibbler7 and Karlin Medium 2.535 -4.411 No, F:tape measure protein, SIF-BLAST:Tape Measure Protein Supported by NCBI BLAST Arcadia ASR79986.1 0.99 0, Tape Measure Protein Supported by PhagesDB BLAST Nason gp22 0.99 0, tape measure domain supported by a CD found from the TIGR database gp TIGR02675 NA 0.00000202, , SIF-HHPred:Phage-related tail protein supported by COG gpN/A COG5283 0.2708 1, SIF-Syn:NKF, upstream gene is NKF, downstream gene is NKF, just like in phage Nason

Conclusions:

It can be concluded that gene 18 codes for a minor tail protein as does gene 19. It can also be concluded that gene 20 codes for a tape measure protein. There were no significant challenges in annotating these genes.

Next Steps:

The next step will be to fix any errors found in the annotation of NapoleonB in order for it to be submitted to PhagesDB and subsequently NCBI.