Part:BBa_K1172902:Design

alanine racemase (''alr'') under the control of the P_''tac'' promoter

Assembly Compatibility:

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

  Illegal NheI site found at 385
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 309
  Illegal BamHI site found at 1011
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 427
  Illegal AgeI site found at 727
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 184
  

Design Notes

Amplification with Gibson-primer:

• Amplification of the alanin racemase (alr) under the control of the P_tac promoter:
  

pSB1C3_ptac_alr (70 bp)

pSB1C3_ptac_alr (70 bp)
5'GTTGACAATTAATCATCGGCTCGTATAATACTAGAGAAAGAGGAGAAATACTAGATGCAAGCGGCAACTGTTGTG-3'

pSB1C3_alr_rev (41 bp)

5'-CTGCAGCGGCCGCTACTAGTATTAATCCACGTATTTCATCG-3'

• Amplification of the pSB1C3-Backbone:
  

pSB1C3_alr_suf (41 bp)

5'-CGATGAAATACGTGGATTAATACTAGTAGCGGCCGCTGCAG-3'

pSB1C3_ptac_alr_pre (41 bp)

5'-CTTTCACACATTATACGAGCCGATGATTAATTGTCAACCTAGAAGCGGCCGCGAATT-3'

Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].

Source

• Genomic DNA Escherichia coli K-12
  • Isolation of the whole E. coli genome, see [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation here]