LOCUS AY562546 8255 bp DNA circular SYN 05-APR-2004 DEFINITION Binary Vector pORE_R3, complete sequence. ACCESSION AY562546 VERSION AY562546.1 GI:45862391 KEYWORDS . SOURCE Binary vector pORE_R3 ORGANISM Binary vector pORE_R3 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 8255) AUTHORS Coutu,C.M., Brandle,J., Brown,K., Simmonds,J., Brown,D., Mici,B. and Hegedus,D.D. TITLE pORE: a new series of binary vectors with a removable selectable marker and 21 site MCS JOURNAL Unpublished REFERENCE 2 (bases 1 to 8255) AUTHORS Coutu,C.M., Brandle,J., Brown,K., Simmonds,J., Brown,D., Mici,B. and Hegedus,D.D. TITLE Direct Submission JOURNAL Submitted (01-MAR-2004) Molecular Genetics, Agriculture and Agrifood Canada, 107 Science Place, Saskatoon, Saskatchewan S7N 0X2, Canada FEATURES Location/Qualifiers source 1..8255 /organism="Binary vector pORE_R3" /mol_type="other DNA" /db_xref="taxon:267423" /lab_host="Agrobacterium tumefaciens" terminator 1..253 /note="NOS terminator; polyadenylation signal from nopaline synthase gene of the Agrobacterium Ti plasmid; derived from pBI121 in GenBank Accession Number AF485783" misc_recomb 260..307 /note="FRT site; excision of selectable marker cassette from transformed plants using FLP recombinase" misc_feature 356..3610 /note="derived from pCB301 in GenBank Accession Number AF139061; used SDM to remove NotI, XhoI, EcoRV and SfiI sites" misc_feature 356..516 /note="T-DNA left border" rep_origin 523..1152 /note="RK2 oriV" CDS 1364..2158 /note="neomycin-kanamycin phosphotransferase type III" /codon_start=1 /transl_table=11 /product="NptIII" /protein_id="AAS78971.1" /db_xref="GI:45862392" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLK MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEY EDEQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWE EDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKW YDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" CDS 2457..3605 /note="derived from plasmid RK2" /codon_start=1 /transl_table=11 /product="TrfA" /protein_id="AAS78972.1" /db_xref="GI:45862393" /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVG SMVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKK RKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALF TTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPI TFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLD RGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKL ETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" rep_origin 3663..4543 /note="ColE1; high copy number origin of replication" misc_feature 4549..4711 /note="T-DNA right border" primer_bind 4724..4746 /note="M13/pUC forward" /PCR_conditions="60 degree C annealing temperature" misc_feature 4747..4868 /note="multiple cloning site; selected unique sites: SfoI-SacII-HindIII-NheI-MfeI-PspOMI-AclI-XhoI-XbaI-StuI- BamHI-SmaI-EcoRI-SalI-NotI-ClaI-PstI-SacI" CDS 4874..5593 /note="small, modified green fluorescent protein; removed ClaI, MfeI and NcoI sites; derived from psmGFP in GenBank Accession Number U70495" /codon_start=1 /transl_table=11 /product="smGFP" /protein_id="AAS78973.1" /db_xref="GI:45862394" /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT LKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKN GIKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH MVLLEFVTAAGITHGMDELYK" primer_bind complement(5616..5638) /note="M13/pUC reverse" /PCR_conditions="60 degree C annealing temperature" terminator 5645..5897 /note="NOS terminator; polyadenylation signal from nopaline synthase gene of the Agrobacterium Ti plasmid; derived from pBI121 in GenBank Accession Number AF485783" misc_recomb 5904..5951 /note="FRT site; excision of selectable marker cassette from transformed plants using FLP recombinase" promoter 5968..7444 /note="At4g15440 promoter; hydroperoxide lyase promoter; constitutive; high level expression of transgenes in dicots; active in Arabidopsis, tobacco, cauliflower, soybean, alfalfa, peach, white spruce, wheat, and grape; in Arabidopsis, expression has been detected in all tissues but the petals, style, and pollen; removed ClaI, SfoI, XhoI and MfeI sites" CDS 7453..8247 /note="neomycin phosphotransferase II; removed SfoI, PstI, and NcoI sites; repaired reduction-in-function mutation; derived from pBI121 in GenBank Accession Number AF485783" /codon_start=1 /transl_table=11 /product="NptII" /protein_id="AAS78974.1" /db_xref="GI:45862395" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" ORIGIN 1 gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg 61 atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc 121 atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac 181 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct 241 atgttactag atccctaggg aagttcctat tccgaagttc ctattctctg aaaagtatag 301 gaacttcttt gcgtattggg cgctcttggc ctttttggcc accggtcgta cggttaaaac 361 caccccagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt 421 ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca 481 caaaatcacc actcgataca ggcagcccat cagtccacta gacgctcacc gggctggttg 541 ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa acgccgtcga 601 agccgtgtgc gagacaccgc agccgccggc gttgtggata cctcgcggaa aacttggccc 661 tcactgacag atgaggggcg gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt 721 tgacagatga ggggcaggct cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa 781 tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat gatgtggaca agcctgggga 841 taagtgccct gcggtattga cacttgaggg gcgcgactac tgacagatga ggggcgcgat 901 ccttgacact tgaggggcag agtgctgaca gatgaggggc gcacctattg acatttgagg 961 ggctgtccac aggcagaaaa tccagcattt gcaagggttt ccgcccgttt ttcggccacc 1021 gctaacctgt cttttaacct gcttttaaac caatatttat aaaccttgtt tttaaccagg 1081 gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc 1141 tcccggcccg ctctcgcgtt ggcagcatca cccataattg tggtttcaaa atcggctccg 1201 tcgatactat gttatacgcc aactttgaaa acaactttga aaaagctgtt ttctggtatt 1261 taaggtttta gaatgcaagg aacagtgaat tggagttcgt cttgttataa ttagcttctt 1321 ggggtatctt taaatactgt agaaaagagg aaggaaataa taaatggcta aaatgagaat 1381 atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat 1441 gtctcctgct aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac 1501 ggacagccgg tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg 1561 gctggaagga aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag 1621 caatctgctc atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca 1681 aagccctgaa aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga 1741 catatcggat tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt 1801 actgaataac gatctggccg atgtggattg cgaaaactgg gaagaagaca ctccatttaa 1861 agatccgcgc gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt 1921 ttcccacggc gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt 1981 tattgatctt gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg 2041 gtcgatcagg gaggatattg gggaagaaca gtatgtcgag ctattttttg acttactggg 2101 gatcaagcct gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagta 2161 cctagatgtg gcgcaacgat gccggcgaca agcaggagcg caccgacttc ttccgcatca 2221 agtgttttgg ctctcaggcc gaggcccacg gcaagtattt gggcaagggg tcgctggtat 2281 tcgtgcaggg caagattcgg aataccaagt acgagaagga cggccagacg gtctacggga 2341 ccgacttcat tgccgataag gtggattatc tggacaccaa ggcaccaggc gggtcaaatc 2401 aggaataagg gcacattgcc ccggcgtgag tcggggcaat cccgcaagga gggtgaatga 2461 atcggacgtt tgaccggaag gcatacaggc aagaactgat cgacgcgggg ttttccgccg 2521 aggatgccga aaccatcgca agccgcaccg tcatgcgtgc gccccgcgaa accttccagt 2581 ccgtcggctc gatggtccag caagctacgg ccaagatcga gcgcgacagc gtgcaactgg 2641 ctccccctgc cctgcccgcg ccatcggccg ccgtggagcg ttcgcgtcgt ctcgaacagg 2701 aggcggcagg tttggcgaag tcgatgacca tcgacacgcg aggaactatg acgaccaaga 2761 agcgaaaaac cgccggcgag gacctggcaa aacaggtcag cgaggccaag caagccgcgt 2821 tgctgaaaca cacgaagcag cagatcaagg aaatgcagct ttccttgttc gatattgcgc 2881 cgtggccgga cacgatgcga gcgatgccaa acgacacggc ccgctctgcc ctgttcacca 2941 cgcgcaacaa gaaaatcccg cgcgaggcgc tgcaaaacaa ggtcattttc cacgtcaaca 3001 aggacgtgaa gatcacctac accggcgtcg agctgcgggc cgacgatgac gaactggtgt 3061 ggcagcaggt gttggagtac gcgaagcgca cccctatcgg cgagccgatc accttcacgt 3121 tctacgagct ttgccaggac ctgggctggt cgatcaatgg ccggtattac acgaaggccg 3181 aggaatgcct gtcgcgccta caggcgacgg cgatgggctt cacgtccgac cgcgttgggc 3241 acctggaatc ggtgtcgctg ctgcaccgct tccgcgtcct ggaccgtggc aagaaaacgt 3301 cccgttgcca ggtcctgatc gacgaggaaa tcgtcgtgct gtttgctggc gaccactaca 3361 cgaaattcat atgggagaag taccgcaagc tgtcgccgac ggcccgacgg atgttcgact 3421 atttcagctc gcaccgggag ccgtacccgc tcaagctgga aaccttccgc ctcatgtgcg 3481 gatcggattc cacccgcgtg aagaagtggc gcgagcaggt cggcgaagcc tgcgaagagt 3541 tgcgaggcag cggcctggtg gaacacgcct gggtcaatga tgacctggtg cattgcaaac 3601 gctagggcct tgtggggtca gttccggctg ggggttcagc agccagcgct ttactgagat 3661 cctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 3721 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 3781 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 3841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 3901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 3961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4081 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4261 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 4321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 4441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 4501 tggtcatgag attatcaaaa aggatcttca cctagatcct tttggatctc ctgtggttgg 4561 catgcacata caaatggacg aacggataaa ccttttcacg cccttttaaa tatccgatta 4621 ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc aaacactgat 4681 agtttaaact gaaggcggga aacgacaatc tgctagtgga tctcccagtc acgacgttgt 4741 aaaacgggcg ccccgcggaa agcttgctag ccaattgggg cccaacgttc tcgagttttt 4801 ctagaaggcc ttggatccac ccgggagaat tcgtcgactt tgcggccgca tcgatactgc 4861 aggagctcca aaaatggcga gtaaaggaga agaacttttc actggagttg tcccaattct 4921 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg 4981 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt 5041 tccttggcca acacttgtca ctactttctc ttatggtgtt caatgctttt caagataccc 5101 agatcatatg aagcggcacg acttcttcaa gagcgccatg cctgagggat acgtgcagga 5161 gaggaccatc tctttcaagg acgacgggaa ctacaagaca cgtgctgaag tcaagtttga 5221 gggagacacc ctcgtcaaca ggatcgagct taagggaatt gatttcaagg aggacggaaa 5281 catcctcggc cacaagttgg aatacaacta caactcccac aacgtataca tcacggcaga 5341 caaacaaaag aatggaatca aagctaactt caaaattaga cacaacattg aagatggaag 5401 cgttcaacta gcagaccatt atcaacaaaa tactcctatt ggcgatggcc ctgtcctttt 5461 accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag 5521 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga 5581 actatacaaa taaggtacca aaaactagtg atatccctgt gtgaaattgt tatccgctac 5641 gcgtgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct 5701 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta 5761 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta 5821 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc 5881 atctatgtta ctagatccca tgggaagttc ctattccgaa gttcctattc tctgaaaagt 5941 ataggaactt cagcgatcgc agacgtcaac gtggatactt ggcagtggtt acttggcttt 6001 tcctttattt tcttttggac ggaagcggtg gttactttgt cacacattta aaaaaacacg 6061 tgtttctcac ttttttctat tcccgtcaca aacaatttta agaaagatcc atctatcgtg 6121 atctttctat caaacaaaag aaaaaaggtc ttcatagtaa cgctacaaca tcaaatatgt 6181 ggttgctctg acatcagtcg ggaaaataag gatatggcgg cattggccac atctattggg 6241 gtcccaactt cctttcacaa aaaaattaaa ttgggtgtcc caacttttat ctttgatata 6301 gtgacatgag tatcgggagc attggacaat ggataaaatg agaactaaaa aaattctggt 6361 taatttttga tcattgttat ttaaaaggtt attttatcta taatctaccc atattgatca 6421 gttttattta aatttgttta gctaccgctc cacgagagag atcctcatct taaaaatgga 6481 atatggaaat tacacacgac cccaaaagta tattttttct ctggagaatg ctatttagag 6541 ctttgactat atggtctgaa ttagaaagac gggaaataaa atctgctaag tgatataagc 6601 tctaagtagg cgatgtgtga tggagaacac cttttcttta acagtcttca tgttttacag 6661 attcgcgaac ttcgaatatc cctatacggt ctgtctaacc ctcgtgtgtc ttttgagtcc 6721 aagataaagg ccattattga gtaacataga catgctggaa tccaaccatt gaagtcacaa 6781 ctgtccatgt agattctttg gagaatctga aaagtcttaa taaaggtggt gtttcaaaga 6841 aaacaaaaca aatgagttaa gaaaaaaaaa tatcatgtag tggtcgagta ttatgttatt 6901 tattgtgtag ctaccaatct ttattcttta aatctgacat aaaatgctac aaacttttta 6961 cctcgtctat agccccaaaa aacctaacca cggttctaaa accacacaca gtgattttgg 7021 ttgacgacaa tgcctctcct tcctcaaaac gatttattta cattttttaa atcaaatgtt 7081 acattttata ccataattaa gtctttttac agaatactta gatggaagag atgtataaaa 7141 aaggaggaaa ttgtaaaaaa catatttcga tcaattaaac caggattcat aaaaatataa 7201 gtatatatat aaatgatgtt tcgtttagcg atgaacttca ctcatatgat aatacttaac 7261 aatataagta cataaaaaat aaaataaaat taattgttta cgaaaagtct acaaatactg 7321 catgtataat taatgttctc tttatttatt tatttatacc ttaccaagat atatctataa 7381 ccgcatagaa atagaaggcg aagagataat ttccaaaaac aagaaaaacc tctaagctca 7441 aaagggccgg ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 7501 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 7561 ttccggctgt cagcgcaggg gaggccggtt ctttttgtca agaccgacct gtccggtgcc 7621 ctgaatgaac ttcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 7681 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 7741 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 7801 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 7861 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 7921 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 7981 cgcatgcccg acggcgagga tctcgtcgtg actcatggcg atgcctgctt gccgaatatc 8041 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 8101 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 8161 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 8221 tatcgccttc ttgacgagtt cttctgaggc gcgcc //