Part:BBa_I737020

PoPS->Agarase (Digests agar)

This part is Beta-Agarase cloned out of Pseudoalteromonas atlantica T6C from the ATCC. Pseudoalteromonas atlantica is a gram-negative, motile, chemoorganotroph which lives in sea water. We made a series of primers and cloned agarase using a TOPO cloning kit. We then cloned it into pSB1A2 using XbaI and SpeI after amplifying with primers that add the BioBrick prefix and suffix. Currently the gene has a EcorRI and a PstI site which limits how it can be assembled. It does however have the proper BioBrick prefix and suffix and can be assembled using XbaI and SpeI. Unfortunately this is not directional.

This part has been sequenced and confirmed to be the beta-agarase from Pseudoalteromonas atlantica T6C. The sequence (from genome of T6C) can be found here: http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=109896332&db=Nucleotide&from=3202543&to=3204984&view=gbwithparts&RID=J2F5YTKG01R

Sequencing results can be found on the University of Calgary iGEM 2007 Wiki.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1861
  Illegal PstI site found at 284
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1861
  Illegal NheI site found at 106
  Illegal NheI site found at 901
  Illegal PstI site found at 284
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1861
  Illegal XhoI site found at 1964
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1861
  Illegal PstI site found at 284
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1861
  Illegal PstI site found at 284
  Illegal AgeI site found at 368
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1319
  Illegal SapI.rc site found at 1717
  Illegal SapI.rc site found at 1795