Coding

Part:BBa_K174000:Design

Designed by: The Newcastle 2009 iGEM team   Group: iGEM09_Newcastle   (2009-09-26)
Revision as of 19:04, 26 September 2009 by Goksel (Talk | contribs) (→‎Construction)

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

Treat Plasmid part BBa_J04450 (pSB1AT3) and our construct with EcoRI and SpeI

Newcastle 2009 sspB 1.png

Ligation of the backbone and the sspB insert

Newcastle 2009 sspB 2.png