9.2.2 CESG CESG_1 John Markley

Dept. of Biochemistry Univ. of Wisconsin Madison WI 53706

USA markley@nmrfam.wisc.edu The Center for Eukaryotic Structural Genomics CESG Principal Investigator CESG_2 George Phillips Jr.

Dept. of Biochemistry Univ. of Wisconsin Madison WI 53706

USA phillips@biochem.wisc.edu CESG CESG co-director Selection_1 Target Selection Selection of targets from eukaryotes. Provide detailed description of your protocol here... Cloning_1 Free Ligation Cloning Cloning into vector XXXXXX Provide detailed description of your protocol here... GO.22030 PSI 1 PSI 2 biomedical Human Metastasis community nominated Amidohydrolase requested by Frank Raushel, Texas University cloned TX1001 a similar sequence in mouse TX900 a similar sequence in rat ATGGCTTTTTCCGATGATCTCCCACCACCACCACGTGTGAACGATCATGTGAAGACGAGAAGAAACAGAAAAAGAAGAACGATCAAGACGAAGCAACTTGAAGAATTGATAAGCACCGCCACAAGAGCCGCTCATGTTGCAAGAGACAAAGGGTTTTACATAGTCTCTCCGGAAGCCATACAATGCGTTGAGATCCTTAGACATATGCGAACCTTACCTCTAAACGCTCGTCTTATTACCAAAACCGACGGTTTACGCGTTCTTCTATTTCTCTCTAAGAACGGTAACCCTAAAATCAGGTCTGAGTCAAAGGCCGTCATTGATCACTGGAAAAGCATTCTCCATACCAAAGTTCATTAA predicted dna dna full ORF MAFSDDLPPPPRVNDHVKTRRNRKRRTIKTKQLEELISTATRAAHVARDKGFYIVSPEAIQCVEILRHMRTLPLNARLITKTDGLRVLLFLSKNGNPKIRSESKAVIDHWKSILHTKVH predicted protein protein full ORF Amidohydrolase http://www.doe-mbi.ucla.edu/TB/PUBLIC/qs/qsearch.php?dowork=Rv2031c If you have something to say about this target say it now.. Arabidopsis thaliana PFAM PF00025 Genbank 189676547 BIG BIG_125 CESG_1 CESG_2 1 2008-01-16 CESG_1 cloned CESG selected 2005-07-19 CESG cloned 2005-07-25 AAACCCCAAGCAGCACGTCCCCTTGGATGAGTACAGCGCAAACCTGAGGGACATGGTGCAGTACCTGAGGTCTGTGGACGTCCCCAGGGAGCGGGTCATCCTTATCACGCCACCCCCGCTCTGCGAAGCCGCCTGGGAGAAGGAGTGCGTCCTGAAGGGTTGCAAATTAAACCGCCTGAACTCAGTTGTTGGTGAATATGCAAATGCGTGTTTACAAGTAGCCCGAGACTGTGGGACTGATGTCCTTGACCTGTGGACCCTGATGCAGAAGGACAGCCAGGACTTCTCATCCTATTTATCGGATGGCCTGCATTTATCACCAATGGGAAATGAGTTTCTGTTCTTAAACCTCTGTCCACTGCTGGACAAGAAGGTCTCATCTCTGCCGTGGCTGCTTCCTTACTGGAAGGATGTGGAGGAAGCAAAGCCCGAGCTGAGTCTGCTGGGAGATGGAGACTATTAG dna MPRLIRKGPGMENPVAVTIFFGANDSSLKDENPKQHVPLDEYSANLRDMVQYLRSVDVPRERVILITPPPLCEAAWEKECVLKGCKLNRLNSVVGEYANACLQVARDCGTDVLDLWTLMQKDSQDFSSYLSDGLHLSPMGNEFLFLNLCPLLDKKVSSLPWLLPYWKDVEEAKPELSLLGDGDY protein valine 7 replaced by proline five terminal residues cleaved Selection_1 selection host: Escherichia Coli strain: BL21magic cloning vector: pMCSG7 Cloning_1 cloning host: Escherichia Coli strain: BL21magic cloning vector: pMCSG7 2 2008-01-17 CESG_1 work stopped CESG selected 2005-07-19 CESG work stopped 2005-07-25 cloning failed poor transformation AAACCCCAAGCAGCACGTCCCCTTGGATGAGTACAGCGCAAACCTGAGGGACATGGTGCAGTACCTGAGGTCTGTGGACGTCCCCAGGGAGCGGGTCATCCTTATCACGCCACCCCCGCTCTGCGAAGCCGCCTGGGAGAAGGAGTGCGTCCTGAAGGGTTGCAAATTAAACCGCCTGAACTCAGTTGTTGGTGAATATGCAAATGCGTGTTTACAAGTAGCCCGAGACTGTGGGACTGATGTCCTTGACCTGTGGACCCTGATGCAGAAGGACAGCCAGGACTTCTCATCCTATTTATCGGATGGCCTGCATTTATCACCAATGGGAAATGAGTTTCTGTTCTTAAACCTCTGTCCACTGCTGGACAAGAAGGTCTCATCTCTGCCGTGGCTGCTTCCTTACTGGAAGGATGTGGAGGAAGCAAAGCCCGAGCTGAGTCTGCTGGGAGATGGAGACTATTAG dna MPRLIRKGPGMENPVAVTIFFGANDSSLKDENPKQHVPLDEYSANLRDMVQYLRSVDVPRERVILITPPPLCEAAWEKECVLKGCKLNRLNSVVGEYANACLQVARDCGTDVLDLWTLMQKDSQDFSSYLSDGLHLSPMGNEFLFLNLCPLLDKKVSSLPWLLPYWKDVEEAKPELSLLGDGDY protein valine 7 replaced by proline five terminal residues cleaved Selection_1 selection host: Escherichia Coli strain: BL21magic cloning vector: pMCSG7 Cloning_1 cloning