Coding
Part:BBa_K174000:Design
Designed by: The Newcastle 2009 iGEM team Group: iGEM09_Newcastle (2009-09-26)
498 bp long sspB CDS is amplified by PCR with dangling end primers with EcoRI-NotI-XbaI restriction site at 5’ and SpeI at 3’ and inserted into a Biobrick compatible vector. The sequence is taken from E. coli strain DH5 alpha. Genbank accession number for E. coli MG1655 strain is NC_000913.2
Forward primer used: GATCTG-GAATTCGCGGCCGCTTCTAG-ATGGATTTGTCACAGCTAAC (Clamp sequence - Standard Biobrick prefix - first 20 base from the Biobrick)
Reverse primer used: TGTGAC-ACTAGTA-TTACTTCACAACGCGTAATGC (Clamp sequence - SpeI site - last 21 base from the Biobrick)
Construction
Plasmid part BBa_J04450 (pSB1AT3) and our construct were treated with EcoRI and SpeI
The backbone and the sspB insert were then ligated
