Part:BBa_K896002:Design

Dsr (sulfite reductase)

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1636
  Illegal EcoRI site found at 4648
  Illegal PstI site found at 826
  Illegal PstI site found at 3838
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1636
  Illegal EcoRI site found at 4648
  Illegal PstI site found at 826
  Illegal PstI site found at 3838
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1636
  Illegal EcoRI site found at 4648
  Illegal BglII site found at 1091
  Illegal BglII site found at 4103
  Illegal BamHI site found at 3007
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1636
  Illegal EcoRI site found at 4648
  Illegal PstI site found at 826
  Illegal PstI site found at 3838
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1636
  Illegal EcoRI site found at 4648
  Illegal PstI site found at 826
  Illegal PstI site found at 3838
  Illegal NgoMIV site found at 1741
  Illegal NgoMIV site found at 4753
  Illegal AgeI site found at 406
  Illegal AgeI site found at 1261
  Illegal AgeI site found at 3418
  Illegal AgeI site found at 4273
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

1. Endogenious EcoRI sites are at 1636bp and 4143bp, and PstI sites are at 826bp and 3833bp.

2. Endogenious BglII sites are at 1091bp and 4098bp, and NgoMIV sites are at 1741bp and 4748bp.

3. Endogenious AgeI sites are at 406bp, 1261bp, 3413bp, 4268bp.

4. We used BamHI(at 3008bp) to combine DsrI and DsrII gene, and produced Dsr gene.

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

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

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

8. Insert: Dsr, cut with EcoRI and XbaI

Source

Wildtype sulfite reductase which is originatal from Desulfovibrio desulfuricans w/o engineered.