Part:BBa_K896001:Design

CysI (Sulfite reductase)

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 655
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  Illegal NgoMIV site found at 394
  Illegal NgoMIV site found at 580
  Illegal NgoMIV site found at 722
  Illegal AgeI site found at 94
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 130
  Illegal BsaI site found at 1452
  

Design Notes

1. Endogenious EcoRI site is at 655bp, and PstI is at 121bp.

2. Instead of mutant ctting site, we created a smart pSB1C3 plasmid (MfeI-XbaI -pSB1C3-SbfI-SpeI)for CysI cloning.

3. Taking advantage of MfeI and EcoRI are compatible; SbfI and XbaI are compatible.

4. Vector: MfeI-XbaI -pSB1C3-SbfI-SpeI, cut with MfeI and SbfI

5. Insert: CysI, cut with EcoRI and XbaI

Source

Wildtype sulfite reductase which is originatal from Pseudomonas aeruginosa PAOI w/o engineered.

References

1.[http://www.wikigenes.org/e/gene/e/878581.html cysI - sulfite reductasePseudomonas aeruginosa PAO1.]

2.A novel reduced flavin mononucleotide-dependent methanesulfonate sulfonatase encoded by the sulfur-regulated msu operon of Pseudomonas aeruginosa. Kertesz, M.A., Schmidt-Larbig, K., Wüest, T. J. Bacteriol. (1999)

3.Regulation of the sulfate starvation response in Pseudomonas aeruginosa: role of cysteine biosynthetic intermediates. Hummerjohann, J., Küttel, E., Quadroni, M., Ragaller, J., Leisinger, T., Kertesz, M.A. Microbiology (Reading, Engl.) (1998)

4.Mapping of cysteine genes on the chromosome of Pseudomonas aeruginosa PAO. Brandt, R., Günther, E., Herrmann, H. Mol. Gen. Genet. (1984)