Purpose: The purpose of this lab is to begin annotating NaopleonB, specifically gene 93.

Tools/Procedures:

Tools:

• DNA Master
• NCBI
• PhagesDB
• HHpred

Procedures:

1. A GeneMark complete sequence was performed on NapoleonB.
2. Using the information from DNA Master and Starterator, the start and stop were determined for gene 93.
3. A BLAST search was completed for gene 93 of NapoleonB on PhagesDB and NCBI.
4. The protein sequence for gene 93 was also run through HHpred.
5. Based on these results, the annotation for gene 93 was completed and added to Phage Notes for NapoleonB.
6. The following annotation was recorded: SSC:52622 – 52879, CP:Yes, SCS:Both, ST:SA, BLAST-Start:Aligns with Arthribacterphage Mudcat gp87 NCBI BLAST q1:s1 0.98 2E-53, Aligns with Arthrobacter phage Xenomorph gp88 PhagesDB BLAST q1:s1 1 3E-48, Gap:129bp gap, LO:Yes, RBS:Kibbler7 and Karlin Medium 2.802 -4.576 Yes, F:Nicotinamate Mononucleoside Transporter, SIF-BLAST:NKF, SIF-HHPred:Nicotinamide mononucleotide transporter supported by PFam gpNA PF04973.12 0.9302 , SIF-Syn:NKF
7. The annotations were saved.

Results:
The annotation for gene 93 of NapoleonB was completed. The function for this gene is not certain at this point. The HHpred search matched with the function Nicotinamide mononucleotide transporter, however, this function is not an official function on the SEA-PHAGES list.
Annotation:
SSC:52622 – 52879, CP:Yes, SCS:Both, ST:SA, BLAST-Start:Aligns with Arthribacterphage Mudcat gp87 NCBI BLAST q1:s1 0.98 2E-53, Aligns with Arthrobacter phage Xenomorph gp88 PhagesDB BLAST q1:s1 1 3E-48, Gap:129bp gap, LO:Yes, RBS:Kibbler7 and Karlin Medium 2.802 -4.576 Yes, F:Nicotinamate Mononucleoside Transporter, SIF-BLAST:NKF, SIF-HHPred:Nicotinamide mononucleotide transporter supported by PFam gpNA PF04973.12 0.9302 , SIF-Syn:NKF

Conclusion:
Annotations were completed for NapoleonB gene 93 in this lab, but the function of the gene is not yet determined. A possible function was found using HHpred, but as this function is not on the official function list of SEA-PHAGES, it cannot yet be called the function. Further investigation will need to be done to determine the function of this gene. Continuing annotations will lead to a better understanding of each of the resources used and will help in making more accurate calls throughout the annotation process.

Future Work:
Future work will include finishing annotations for NapoleonB, specifically genes 1-4. This will be done in the next lab.