Part:BBa_K851004

aminoglycoside antibiotic resistance Sm+ Spc+

The Ω Cassette provides resistance to Spectinomycin and Streptomycin; it was obtained from plasmid pHP45Ω [1] as described in [2].

The aminoglycoside antibiotic resistance gene (aadA +)[3,4] of pHP45 was originally carried on a 1.7-kb PvuII-HindIII fragment from the R100.1 plasmid[5]. Plasmid pHP45Ω was constructed in Pierre Prentki and Henry M. Krisch, 1984 [5] as stated in NCBI[6] with this fragment. The addA sequence can be found in [7, 8 ], it is confirmed to be the one contained in pHP45Ω[9] and the complete sequence of pHP45Ω plasmid can be found in [10, 11].

Hawaii 2008 iGEM team [12] attempted before to construct a similar cassette [13] but didn’t succeed. For iGEM UNAM Genomics México 2012 project [14], the Ω Cassette was used in the design of an OR logic gate[15] using a recently described new type of communication system between Bacillus Subtilis cells called Nanotubes[16].

REFERENCES

[1] http://www.shigen.nig.ac.jp/ecoli/strain/cvectorDetailAction.do?cvectorId=147
[2] http://2012.igem.org/Team:UNAM_Genomics_Mexico/Notebook/ANDSugar
[3] http://www.ihop-net.org/UniPub/iHOP/bng/174343.html
[4] http://www.wikigenes.org/e/gene/e/1238687.html
[5] Pierre Prentki and Henry M. Krisch (1984). In vitro insertional mutagenesis with a selectable DNA fragment. Gene, 29:303-313
[6] http://www.ncbi.nlm.nih.gov/nuccore/K02163
[7] http://www.uniprot.org/uniprot/P0AG05
[8] http://www.ncbi.nlm.nih.gov/sites/entrez?db=gene&term=1446561
[9] Nancye C. Clark, Ørjan Olsvik, Jana M. Swenson, Carol A. Spiegel and Fred C. Tenover(1999) Detection of a Streptomycin/Spectinomycin Adenylyltransferase Gene ( aadA) in Enterococcus faecalis. Antimicrob. Agents Chemother. 43(1):157.
[10] http://www.ncbi.nlm.nih.gov/nuccore/207855?report=fasta
[11] Pierre Prentki, Anna Bind and Andrée Epstein (1991). Plasmid vectors for selecting ISI-promoted deletions in cloned DNA: sequence analysis of the omega interposon. Gene, 103:17-23.
[12] http://2008.igem.org/Team:Hawaii
[13] http://2008.igem.org/Team:Hawaii/Construction_of_Omega_Interposon_BioBrick
[14] http://2012.igem.org/Team:UNAM_Genomics_Mexico
[15] Link con la página de descripción general del sistema o el de la parte del OR
[16] Dubey GP, Ben-Yehuda S. (2011) Intercellular nanotubes mediate bacterial communication. Cell.;144(4) :590-600

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 5
  Illegal BamHI site found at 2060
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1524
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 499