Team:UCL/Lab AzoReducateCloning Portal

BioBrick and Primer Design

Two Azoreductase enzymes will be cloned from Bacilus subtilis (strain 168) and Pseudomonas Aeruginosa (PA01) using DNA PCR amplification.

Suitable forward and reverse primers were designed for each enzymes using the following strategy.

The protein coding sequences for each enzyme were obtained from ?.

ATG TCT ACA GTT TTA TTT GTA AAA TCA AGC GAC CGT ACA GCT GAA GAA GGC GTT TCA ACT AAA CTT TAC GAA
GCT TTC TTA GCT GCT TAT AAA GAA AAC AAC CCT AAT GAT GAA GTG GTT GAA TTA GAT CTT CAT AAG GAA AAC
CTT CCT TAC CTT GGC AGA GAT ATG ATT AAC GGA ACA TTT AAA GCA GGT CAA GGA ATG GAA ATG ACA GAA GAT
GAG AAA AAA CAA GCA GCA ATT GCT GAC AAA TAT CTG AAC CAG TTT GTA AAA GCT GAC AAA GTT GTT TTC GCA
TTC CCG CTT TGG AAC TTC ACA GTG CCA GCA GTG CTT CAT ACT TAT GTT GAT TAT CTG TCT CGC GCA GGC GTT
ACA TTC AAA TAC ACA CAA GAA GGA CCA GTC GGT TTA ATG GGC GGC AAA AAA GTT GCG CTT CTT AAC GCT CGC
GGC GGT GTC TAC TCA GAA GGA CCA ATG GCT GCA CTT GAA ATG TCA TTA AAC TTC ATG AAA ACA GTT CTT GGT
TTC TGG GGT GTT CAA GAC TTG CAC ACA GTT GTC ATC GAA GGA CAT AAC GCA GCA CCT GAT CAA GCG CAA GAA
ATC GTT GAA AAA GGT TTA CAA GAA GCA AAA GAT CTT GCT GCA AAA TTC TAA

The standard BioBrick prefix for protein coding sequences and suffix sequences used in Standard Assembly and belonging to BioBrick RFC 10 were appended to the forward and reverse primers respectively.

BioBrick CDS Prefix

5' GAATTCGCGGCCGCTTCTAG '3

BioBrick Suffix

5' TACTAGTAGCGGCCGCTGCAG '3