cDNA Clone GWD (potato R1)

• Synonyms: R1 starch-granule-bound R1 protein , alpha-glucan water dikinase
• Plasmid: pGEX6P-1-GWD full-length
• Parent Plasmid: pGEX6P-1
• Expressed: GST-GWD full-length
• Genbank: Gene ID: 102577510 NM_001288123.1 → NP_001275052.1
• Tag: GST

Sequence of Insert

ggatccatgagtaattccttagggaataacttgctgtaccagggattcctaacctcaacagtgttggaacataaaagtagaatcagtcctccttgtgttggaggcaattctttgtttcaacaacaagtgatctcgaaatcacctttatcaactgagtttcgaggtaacaggttaaaggtgcagaaaaagaaaatacctatgggaaagaaccgtgctttttctagttctcctcatgctgtacttaccactgatacctcttctgagctagcagaaaagttcagtctagaagggaatattgagctacaggttgatgttaggcctcccacttcaggtgatgtgtcctttgtggattttcaagctacaaatggtagtgataaactgtttttgcactggggggcagtaaagttcggaaaagaaacatggtctcttcctaatgatcgtccagatgggaccaaagtgtacaagaacaaagcacttagaactccatttgttaaatctggctctaactccatcctgagactggagatacgggacactgctatcgaagctattgagtttctcatatacgatgaagcctacgataaatggataaagaataatggtggcaattttcgtgtcaaattgtcaagaaaagagatacgaggcccagatgtttcagttcctgaggagcttgtacagatccaatcatatttgaggtgggagaggaagggaaaacagaattacacccctgagaaagagaaggaggaatatgaggctgctcgaactgagctacaggaggaaatagctcgtggtgcttccatacaggacattcgagcaaggctaacaaaaactaatgataaaagtcaaagcaaagaagagcctcttcatgtaacaaagagtgaaatacctgatgaccttgcccaagcacaagcttacattaggtgggagaaagcaggaaagccgaactatcctccagaaaagcaaattgaagaactcgaagaagcaagaagagaattgcaacttgagcttgagaaaggcattacccttgatgagttgcggaaaaagattacaaaaggggagataaaaactaaggcggaaaagcacgtgaaaagaagctcttttgccgttgaaagaatccaaagaaagaagagagactttgggcagcttattaataagtatccttccagtcctgcagtacaagtacaaaaggtcttggaagaaccaccagccttatctaaaattaagctgtatgccaaggagaaggaggagcagattgatgatccgatccttaataaaaagatctttaaggtcgatgatggggagctactggtactggtagcaaagtcctctgggaagacaaaagtacatatagctacagatctgaatcagccaattactcttcactgggcattatccaaaagtcgtggagagtggatggtaccaccttcaagcatattgcctcctggatcaattattttagacaaggctgccgaaacacctttttccgccagttcttctgatggtctaacttctaaggtacaatctttggatatagtaattgaagatggcaattttgtggggatgccatttgttcttttgtctggtgaaaaatggattaagaaccaagggtcggatttctatgttgacttcagtgctgcatccaaattagcactcaaggctgctggggatggcagtggaactgcaaagtctttactggataaaatagcagatatggaaagtgaggctcagaagtcatttatgcaccggtttaatattgctgctgacttgatagaagatgccactagtgctggtgaacttggttttactggaattcttgtatggatgaggttcatggctacaaggcaactgatatggaacaaaaactataacgtaaaaccacgtgaaataagcaaggctcaggacagacttacagacttgttgcagaatgctttcaccagtcaccctcaataccgtgaaattttgcggatgattatgtcaactgttggacgtggaggtgaaggggatgtaggacagcgaattagggatgaaattttggtcatccagaggaaaaatgactgcaagggtggtatgatggaagaatggcatcagaaattgcataataatactagtcctgatgatgttgtgatctgtcaggcattgattgactacatcaagagtgattttgatcttggtgtttattggaaaaccctgaatgagaacggaataacaaaagagcgtcttttgagttatgaccgtgctatccattctgaaccgaattttagaggagatcaaaagaatggtcttttgcgtgatttaggtcactatatgagaacattgaaggctgttcattcaggtgcagatcttgagtctgctattgcaaactgcatgggctacaaaactgagggagaaggctttatggttggagtccagataaatcctgtatcaggcttgccatctggctttcagggcctcctccattttgtcttagaccatgtggaagataaaaatgtggaaactcttcttgagggattgctagaggctcgtgaggagcttaggcccttgcttctcaaaccaaacaaccgtctaaaggatctgctgtttttggacatagcacttgattctacagttagaacagcagtagaaaggggatatgaagaattgaacaacgctaatcctgagaaaatcatgtacttcatctccctcgttcttgaaaatctcgcactctctgtggacgataatgaagatcttgtttattgcttgaagggatggaatcaagctctttcaatgtccaatggtggagacaaccattgggctttatttgcaaaagctgtacttgacagaatccgtcttgcacttgcaagcaaggcagagtggtaccatcacttattgcagccatctgccgaatatctaggatcaatccttggggtggaccaatgggctttgaacatatttactgaagaaattatacgtgctggatcagcagcttcattatcctctcttcttaatagactcgatcccgtgcttcggaaaactgcaaatctaggaagttggcagattatcagtccagttgaagccgttggatatgttgtcgttgtggatgagttgctttcagttcagaatgaaatctacaagaagcccacgatcttagtagcaaactctgttaaaggagaggaggaaattcctgatggtgctgttgccctgataacaccagacatgccagatgttctttcacatgtttctgttcgagctagaaatgggaaggtttgctttgctacatgctttgatcccaatatattggctgacctccaagcaaaggaaggaaggattttgctcttaaagcctacaccttcagacataatctatagtgaggtgaatgagattgagctccaaagttcaagtaacttggtagaagctgaaacttcagcaacacttagattggtgaaaaagcaatttggtggttgttacgcaatatcagcagatgaattcacaagtgaaatggttggagctaaatcacgtaatattgcatatctgaaaggaaaagtgccttcctcggtgggaattcctacgtcagtagctcttccatttggagtctttgagaaagtactttcagacgacataaatcagggagtggcaaaagagttgcaaattctgacgaaaaaactatctgaaggagacttcagcgctcttggtgaaattcgcacaacgattttagatctttcagcaccagctcaattggtcaaagagctgaaggaaaagatgcagggttctggcatgccttggcctggtgatgaaggtccaaagcggtgggaacaagcatggatggccataaaaaaggtgtgggcttcaaaatggaatgagagagcatacttcagcacaaggaaggtgaaactggatcatgactatctgtgcatggctgtccttgttcaagaaataataaatgctgattatgcatttgtcattcacacaaccaacccatcttccggagacgactcagaaatatatgccgaggtggtcaggggccttggggaaacacttgttggagcttacccaggacgtgctttgagttttatctgcaagaaaaaggatctcaactctCctcaagtgttaggttacccaagcaaaccgatcggccttttcataaaaagatctatcatcttccgatctgattccaatggggaagatttggaaggttatgccggtgctggcctctacgacagtgtaccaatggatgaggaggaaaaagttgtaattgattactcttccgacccattgataactgatggtaacttccgccagacaatcctgtccaacattgctcgtgctggacatgctatcgaggagctatatggctctcctcaagacatcgagggtgtagtgagggatggaaagatttatgtcgttcagacaagacctcagatgtgagcggccgc

Amino Acid Sequence

MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLEVLFQGPLGSMSNSLGNNLLYQGFLTSTVLEHKSRISPPCVGGNSLFQQQVISKSPLSTEFRGNRLKVQKKKIPMGKNRAFSSSPHAVLTTDTSSELAEKFSLEGNIELQVDVRPPTSGDVSFVDFQATNGSDKLFLHWGAVKFGKETWSLPNDRPDGTKVYKNKALRTPFVKSGSNSILRLEIRDTAIEAIEFLIYDEAYDKWIKNNGGNFRVKLSRKEIRGPDVSVPEELVQIQSYLRWERKGKQNYTPEKEKEEYEAARTELQEEIARGASIQDIRARLTKTNDKSQSKEEPLHVTKSEIPDDLAQAQAYIRWEKAGKPNYPPEKQIEELEEARRELQLELEKGITLDELRKKITKGEIKTKAEKHVKRSSFAVERIQRKKRDFGQLINKYPSSPAVQVQKVLEEPPALSKIKLYAKEKEEQIDDPILNKKIFKVDDGELLVLVAKSSGKTKVHIATDLNQPITLHWALSKSRGEWMVPPSSILPPGSIILDKAAETPFSASSSDGLTSKVQSLDIVIEDGNFVGMPFVLLSGEKWIKNQGSDFYVDFSAASKLALKAAGDGSGTAKSLLDKIADMESEAQKSFMHRFNIAADLIEDATSAGELGFTGILVWMRFMATRQLIWNKNYNVKPREISKAQDRLTDLLQNAFTSHPQYREILRMIMSTVGRGGEGDVGQRIRDEILVIQRKNDCKGGMMEEWHQKLHNNTSPDDVVICQALIDYIKSDFDLGVYWKTLNENGITKERLLSYDRAIHSEPNFRGDQKNGLLRDLGHYMRTLKAVHSGADLESAIANCMGYKTEGEGFMVGVQINPVSGLPSGFQGLLHFVLDHVEDKNVETLLEGLLEAREELRPLLLKPNNRLKDLLFLDIALDSTVRTAVERGYEELNNANPEKIMYFISLVLENLALSVDDNEDLVYCLKGWNQALSMSNGGDNHWALFAKAVLDRIRLALASKAEWYHHLLQPSAEYLGSILGVDQWALNIFTEEIIRAGSAASLSSLLNRLDPVLRKTANLGSWQIISPVEAVGYVVVVDELLSVQNEIYKKPTILVANSVKGEEEIPDGAVALITPDMPDVLSHVSVRARNGKVCFATCFDPNILADLQAKEGRILLLKPTPSDIIYSEVNEIELQSSSNLVEAETSATLRLVKKQFGGCYAISADEFTSEMVGAKSRNIAYLKGKVPSSVGIPTSVALPFGVFEKVLSDDINQGVAKELQILTKKLSEGDFSALGEIRTTILDLSAPAQLVKELKEKMQGSGMPWPGDEGPKRWEQAWMAIKKVWASKWNERAYFSTRKVKLDHDYLCMAVLVQEIINADYAFVIHTTNPSSGDDSEIYAEVVRGLGETLVGAYPGRALSFICKKKDLNSPQVLGYPSKPIGLFIKRSIIFRSDSNGEDLEGYAGAGLYDSVPMDEEEKVVIDYSSDPLITDGNFRQTILSNIARAGHAIEELYGSPQDIEGVVRDGKIYVVQTRPQM*

• Vector Type: Bacterial
• Antibiotic: Amp
• Full Snapgene Map: pGEX6P-1-GWD full-length726.54 KB Snapgene files can be opened using software available here: www.snapgene.com
• DU Number: DU71003
• Unit Source: Protein Phosphorylation and Ubiquitylation Unit
• Location: PPU
• MTA: Dundee
• Notes: R1 gene from Solanum tuberosum (potato). N-term GST for bacterial expression.; Chloroplastic. total RNA extracted from 3 week old Desiree (v) leaves.; Silent changes compared to NCBI ref at: ; Pro46 (CCA>CCT), Gly135 (GGG>GGA), Leu420 (CTA>CTT), Leu819 (TTG>TTA), Ser1011 (AGC>AGT), Ala1165 (GCT>GCA), Cys1293 (TGT>TGC), Tyr1339 (TAT>TAC), Asp1412 (GAT>GAC); ; non silent at E66G, K68N, R95G, L447I, A599T, T938I, E1034K, A1312T, T1356P, S1427N; (potatoes are highly heterozygous therefore likely to reflect cultivar variation); ; Asn inserted between Ile381 and Lys382 (all numbers here after Ile381 are cited according to the NCBI ref but will be +1 for this variant after position 381); ; ; NEB builder incorporating RT-PCR 1.7kb product comprising EcoR1 (nt1798)-EcoR1 (nt3534) into EcoR1 cut 65991
• Low Temp: No