Part:BBa_J133653

aadA Cassette + Flanked by TLTerminators (missing RBS)

Streptomycin/Spectinomcin antibiotic resistance cassette with bi-directional 6-frame translational termination. It's an Aminoglycoside 3"-Adenylyltransferase (aadA1) gene generator driven with constitutive P(Lac)IQ promoter, B0034 RBS, flanked with bidirectional translational terminators BBa_J133422 (prefix) and BBa_B0042 (postfix). The part was originally designed with an B0034 RBS, however sequencing the physical construct revealed that the slow growing surviving clone was missing the RBS and missing one of the flanking BioBrick scar. Instead of trying to reconstruct the part with B0034 RBS, the wt Endy 5'UTR (strong ribosome binding) was placed between the P(Lac)IQ promoter and the aadA1 CDS instead, removing the BioBrick scar between the promoter and CDS all together.

Sequencing also revealed that there is an E81V mutation in the aadA1 CDS, resulting from a GAG to GTG codon difference from its source plasmid pRL1383a (GenBank AF403426.1).

The DNA sequence here is updated to reflect the physical construct, lacking the RBS. See part BBa_J133654 for the version with 5UTR (which includes an Bicistronic RBS designed by Drew Endy).

This is an intermediate construct to be flanked with transcriptional terminators.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 703
  
• 1000
  COMPATIBLE WITH RFC[1000]