Part:BBa_K1758330:Design

Lead repressor under control of constitutive promoter and strong RBS

Assembly Compatibility:

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

  Illegal NheI site found at 465
  Illegal NheI site found at 488
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

We used the codon optimized, synthesized Sequence of pbrR from iGEM Team Brown 2007 ( BBa_I721003) under the control of a constitutive Primer( BBa_I721003). We designed the used primes, in specially split primers, for Gibson Assembly to ligate the constitutive Promoter and pbrR with pSB1C3. For this aim we designed four primers for the generation of homologous overlaps between the synthesized constitutive promoter+ pbrR and the pSB1C3:

cm_rev

TATACGCAAGGCGACAAG

pSB1C3_kPrm_fwd

CTAGGACTGAGCTAGCTGTCAATACTAGTAGCGGCCGCTGCA

pSB1C3_rcnR_rev

GAAAGTCCTTGATTCGTACATCAAATGACTCTAGAAGCGGCCGCGAAT

cm_fwd

CGGCATCAGCACCTTGTC

Source

Synthesized, codon optimized gene, designed after ( BBa_I721003) ralstonia metallidurans synthesized by IDT