Part:BBa_K4730000:Design

Level 1 MHETase plasmid

Assembly Compatibility:

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

  Illegal BamHI site found at 1175
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1010
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 992
  Illegal SapI.rc site found at 1291
  

Design Notes

We had to switch to an inducible promoter instead of our original T7 because having the gene expressed constantly added stress to the cells.

Source

ATG CCT ATC ATT CGT AGT ACC ACT GCA TTG GCA GCG ATC GCC TCC GGG TTC TTT CTG ACG GGG GTT GAT CAC GCG GAT GCC GCT GTT GCG GGA GAT TTC CCA AAC CAG TTC TTA TCC TGT TTT GAT GCG GCG AAT TTG ACC GAA ATT GAG CTG CCC GCA GAT GTC CAA GGC TTC CGC TTA ATT GAA ATT GCA GAA CAT GCC GGC GAT AAA GGC ATG CCT GCA CAT TGT GAA ATT GTC GGA GCT ATC AAT GAC CGC ATC TCT CCT GTG GAT GGT CAA CAT TAC TCA ATT AAA TTC CGC TTG CGC CTG CCG CAA GAT TGG AAT GGC CGC TTC TAC ATG GAA GGA GGT GGT GGA TCA AAC GGT GTC CTG AAG GAT GCG ATG GGC CCC ACC GGC TTG AAT CAG GAA GAC TCC GCT CTG GAA CGC GGC TTC GCC GTT GTC ACT ACT GAC AGC GGC CAC GAC AAC GAC ACA AAC TCA GAC TCA AAC GCC TCT GGG CGC TCA GCT TTC GGC ATG GAC CCA CAG GCT CGC CTG GAC TTC GGT TAC ATG TCT TAT GAC ATT GTT ACA CGT GTT GGC AAA GCA ATC GTC GAA AAG TAT TAC GGC GCG GCT CCG GAA AAG AGT TAT TTT ATC GGG TGT AGC GAA GGG GGT CGT GAA GCG GCT TTG ATG ACT CAG CGC TAC CCA GAC TTG TAC GAC GGC GTC GTG GCA GGT GCA CCA GGT ATC CAT TTC TCG TAT TCT GCT GCG TAT GCA CCA TTC CTT CTT CGT ATT TTC GGT AAT CTT GCA GAA ACT CGT AAT CAG TCT GGG CCC GAC GGA ATC CCT CTG TTA AAT AAA TTA TAC TCA GAT AAT GAT GTT CAA CTT ATC GCC GAT GCG GTA GTA GGG GCA TGT GAC GCC CTT GAC GGG CTT GAG GAT CGC ATG AGC AAT AAT ATC GAG GCG TGC ACA ACG GTC ACA GTC CTG CCC CGT TTA CGT GCT TTG ACA TGT AGT GGG GCG AAG GAA GAA GGA TGT TTG TTA GAA GAT CAG ATC GAT GCT TTC GTT GCG GGA ATG GCA GGG CCG GTA ACG TCC GAC GGT ACG CGT TTG TAT CCT GGT CAC CCA TGG GAT CCC GGA ATT GGA GGT CGT ATC GGG GAC AGT GTC AAT GAT GGA TTT CGT TCC TGG TGG TTC GGC TCG TAC GAT TCG GAC CAA AAT AAC GCA CGT AAA GTC ACC CTG AGC ACT CCG CAA CAT GCC ATG CTT TGG CAA ACG CCT CCG GTT CCA TTA CGT CCA GAT GAA TAC GTA CGC TTT GAG ATG AAT TTC AAC ATT GAT GAA ACG CCT GCG CTG GCT TAT GCA ACG ACT GAT TTA TAT CCC GTG AGT TCA GCC GAA CTT GGT AAC GCT GAT TCG CCA GAC TTG TCA GAT TTC GCA TCG CGC GGG GGG AAG TTA GTC ATC TAT CAC GGG GCA GCC GAT GCA GCC TTC TCA GCA TTA GAT ACG ATC AAA TAC TGG AAT GCC GTG AAT GAA ACA GCT GAC GGT CAA GCG GCG GAT TTT GCT CGT TTG TTC ATT ATT CCC GGG ATG AAC CAT TGC CAA GGT GGT CCC GCC ACA GAT GAC GTT GAT TTA TTG ACT CCG CTT ATG GCG TGG GTT GAG GAT GAT TTG CCA ATC GAA CGT TTG GAG GCT ACG GTA TCT AAC CCG GAT TAT TTT GGT GGC AAG AAC TTG AGT CGC CCC CTT TGC CCG TAT CCC CTG TAT GCC GAG TAT GAC GGA GAA GGC GAT CCA AGT AGT GCT GAG TCC TTC ACC TGC GTT GCT AAC TGA