Template:SBB10 ConstructionFiles ZhenH sbb31

sbb31: CA42 origin of replication Source: pEC22-CA42 Target Sequence: agcacttcagcgcgccgtagcatcgataaacattacgggatggggcgaaactgccatctgttcgaaatgacgcgcaaatgggcttacagggcgattcgtcagggctggccagcattctcacagtggcttgatgccgtgattcagcgtgtcgaaatgtacaacgcatcgcttcccgttccgctttcacctcctgaatgtcgggctattggcaagagtattgcgaaatacacgcacaggaacttcacggcggaaactttcgcacagtatgtggctgatacgcacacgccagaaatacaggccaagagaggcaggaaaggtggcatcgctaaaggcgaagcctacgatgacaagcgtttcatggcgctatgtatgctggagaatggatattctcagaaagctattgcggcgatgttggaggtttctactcgaaccattcgaaactggaaaagcggaaaatagcctatatcagataacagcgcctttctggcgtttttttgagcagtaggtcttttgccg Vector: pBjk2741 Short description: oriCA42 Genbank reference: D30056.1 Family: Origin of Replication

PCR ZZH001 and ZZH002 on pEC22-CA42 (547 bp, EcoRI/BamHI) Sub into pBca9523-Bca1144           (EcoRI/BamHI, 2472+910, L) Product is pBca9523-sbb31         {oriCA42}

ZZH001  Forward oligo for cloning of oriCA42 ccataGAATTCatgAGATCTagcacttcagcgcgccgtag ZZh002  Reverse oligo for cloning of oriCA42 ctgatGGATCCcggcaaaagacctactgctc



JCA Notes
PCR ZZH001 and ZZH002 on pEC22-CA42 (547 bp, EcoRI/BamHI) Sub into pBjk2741-Bca1144           (EcoRI/BamHI, 2472+910, L) Product is pBjk2741-sbb31         {oriCA42}
 * Correct, except should clone into pBjk2741-Bca1144

ZZH001  Forward oligo for cloning of oriCA42 ccataGAATTCatgAGATCTagcacttcagcgcgccgtag ZZh002  Reverse oligo for cloning of oriCA42 ctgatGGATCCcggcaaaagacctactgctc