LOCUS V10 7969 bp ds-DNA circular 18-JUN-2010 DEFINITION Binary Vector pORE_E4, complete sequence. ACCESSION AY562537 VERSION AY562537.1 GI:45862352 KEYWORDS . SOURCE Binary vector pORE_E4 ORGANISM Binary vector pORE_E4 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 8029) 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and Hegedus,D.D. TITLE pORE: a new series of binary vectors with a removable selectable marker and 21 site MCS JOURNAL Unpublished REFERENCE 2 (bases 1 to 8029) AUTHORS Coutu,C.M., Brandle,J., Brown,K., Simmonds,J., Brown,D., Mici,B. and Hegedus,D.D. TITLE Direct Submission JOURNAL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 COMMENT COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers source join(1..4760,5323..7969) /organism="Binary vector pORE_E4" /mol_type="other DNA" /db_xref="taxon:267414" /lab_host="Agrobacterium tumefaciens" /label=source:Binary vector pORE_E4 /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink 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" /label=NOS terminator; polyadenylation signal from nopali /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_recomb 260..307 /note="FRT site; excision of selectable marker cassette from transformed plants using FLP recombinase" /label=FRT site; excision of selectable marker cassette f /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_feature 356..3610 /note="derived from pCB301 in GenBank Accession Number AF139061; used SDM to remove NotI, XhoI, EcoRV and SfiI sites" /label=derived from pCB301 in GenBank Accession Number AF /ApEinfo_fwdcolor=#0922ff /ApEinfo_revcolor=green misc_feature 356..516 /note="T-DNA left border" /label=T-DNA left border /ApEinfo_fwdcolor=#0922ff /ApEinfo_revcolor=green rep_origin 523..1152 /note="RK2 oriV" /label=RK2 oriV /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink CDS 1364..2158 /note="neomycin-kanamycin phosphotransferase type III" /codon_start=1 /transl_table=11 /product="NptIII" /protein_id="AAS78941.1" /db_xref="GI:45862353" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLK MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEY EDEQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWE EDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKW YDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" /label=NptIII /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink CDS 2457..3605 /note="derived from plasmid RK2" /codon_start=1 /transl_table=11 /product="TrfA" /protein_id="AAS78942.1" /db_xref="GI:45862354" /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVG SMVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKK RKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALF TTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPI TFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLD RGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKL ETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" /label=TrfA /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink rep_origin 3663..4543 /note="ColE1; high copy number origin of replication" /label=ColE1; high copy number origin of replication /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_feature 4549..4711 /note="T-DNA right border" /label=T-DNA right border /ApEinfo_fwdcolor=#0922ff /ApEinfo_revcolor=green primer_bind 4724..4746 /note="M13/pUC forward" /PCR_conditions="60 degree C annealing temperature" /label=M13/pUC forward /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_feature 4747..4760 /note="multiple cloning site 1; selected unique sites: SfoI-SacII-HindIII-NheI-MfeI-PspOMI-AclI-XhoI" /label=multiple cloning site 1; selected unique sites: Sf /ApEinfo_fwdcolor=#0922ff /ApEinfo_revcolor=green misc_feature 5323..5329 /note="multiple cloning site 2; selected unique sites: XbaI-StuI-BamHI-SmaI-EcoRI-SalI-NotI-ClaI-PstI-SacI-KpnI- SpeI-EcoRV" /label=multiple cloning site 2; selected unique sites: Xb /ApEinfo_fwdcolor=#0922ff /ApEinfo_revcolor=green primer_bind complement(5330..5352) /note="M13/pUC reverse" /PCR_conditions="60 degree C annealing temperature" /label=M13/pUC reverse /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink terminator 5359..5611 /note="NOS terminator; polyadenylation signal from nopaline synthase gene of the Agrobacterium Ti plasmid; derived from pBI121 in GenBank Accession Number AF485783" /label=NOS terminator; polyadenylation signal from nopali(1) /ApEinfo_label=NOS terminator; polyadenylation signal from nopali /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_recomb 5618..5665 /note="FRT site; excision of selectable marker cassette from transformed plants using FLP recombinase" /label=FRT site; excision of selectable marker cassette f(1) /ApEinfo_label=FRT site; excision of selectable marker cassette f /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink promoter 5682..7158 /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" /label=At4g15440 promoter; hydroperoxide lyase promoter; /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green CDS 7167..7961 /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="AAS78943.1" /db_xref="GI:45862355" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" /label=NptII /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_feature 5269..5318 /label=biobrick MCS /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green promoter 4761..5268 /label=peNTCUP2 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green 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 AGCTTGGGAT CTTCTGCAAG CATCTCTATT TCCTGAAGGT 4801 CTAACCTCGA AGATTTAAGA TTTAATTACG TTTATAATTA CAAAATTGAT TCTAGTATCT 4861 TTAATTTAAT GCTTATACAT TATTAATTAA TTTAGTACTT TCAATTTGTT TTCAGAAATT 4921 ATTTTACTAT TTTTTATAAA ATAAAAGGGA GAAAATGGCT ATTTAAATAC TAGCCTATTT 4981 TATTTCAATT TTAGCTTAAA ATCAGCCCCA ATTAGCCCCA ATTTCAAATT CAAATGGTCC 5041 AGCCCAATTC CTAAATAACC CACCCCTAAC CCGCCCGGTT TCCCCTTTTG ATCCATGCAG 5101 TCAACGCCCA GAATTTCCCT ATATAATTTT TTAATTCCCA AACACCCCTA ACTCTATCCC 5161 ATTTCTCACC AACCGCCACA TAGATCTATC CTCTTATCTC TCAAACTCTC TCGAACCTTC 5221 CCCTAACCCT AGCAGCCTCT CATCATCCTC ACCTCAAAAC CCACCGGAGA ATTCGCGGCC 5281 GCTTCTAGAA TTAGAGGACT AGTAGCGGCC GCTGCAGGct agtgatatcc ctgtgtgaaa 5341 ttgttatccg ctacgcgtga tcgttcaaac atttggcaat aaagtttctt aagattgaat 5401 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta 5461 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg 5521 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta 5581 tcgcgcgcgg tgtcatctat gttactagat cccatgggaa gttcctattc cgaagttcct 5641 attctctgaa aagtatagga acttcagcga tcgcagacgt caacgtggat acttggcagt 5701 ggttacttgg cttttccttt attttctttt ggacggaagc ggtggttact ttgtcacaca 5761 tttaaaaaaa cacgtgtttc tcactttttt ctattcccgt cacaaacaat tttaagaaag 5821 atccatctat cgtgatcttt ctatcaaaca aaagaaaaaa ggtcttcata gtaacgctac 5881 aacatcaaat atgtggttgc tctgacatca gtcgggaaaa taaggatatg gcggcattgg 5941 ccacatctat tggggtccca acttcctttc acaaaaaaat taaattgggt gtcccaactt 6001 ttatctttga tatagtgaca tgagtatcgg gagcattgga caatggataa aatgagaact 6061 aaaaaaattc tggttaattt ttgatcattg ttatttaaaa ggttatttta tctataatct 6121 acccatattg atcagtttta tttaaatttg tttagctacc gctccacgag agagatcctc 6181 atcttaaaaa tggaatatgg aaattacaca cgaccccaaa agtatatttt ttctctggag 6241 aatgctattt agagctttga ctatatggtc tgaattagaa agacgggaaa taaaatctgc 6301 taagtgatat aagctctaag taggcgatgt gtgatggaga acaccttttc tttaacagtc 6361 ttcatgtttt acagattcgc gaacttcgaa tatccctata cggtctgtct aaccctcgtg 6421 tgtcttttga gtccaagata aaggccatta ttgagtaaca tagacatgct ggaatccaac 6481 cattgaagtc acaactgtcc atgtagattc tttggagaat ctgaaaagtc ttaataaagg 6541 tggtgtttca aagaaaacaa aacaaatgag ttaagaaaaa aaaatatcat gtagtggtcg 6601 agtattatgt tatttattgt gtagctacca atctttattc tttaaatctg acataaaatg 6661 ctacaaactt tttacctcgt ctatagcccc aaaaaaccta accacggttc taaaaccaca 6721 cacagtgatt ttggttgacg acaatgcctc tccttcctca aaacgattta tttacatttt 6781 ttaaatcaaa tgttacattt tataccataa ttaagtcttt ttacagaata cttagatgga 6841 agagatgtat aaaaaaggag gaaattgtaa aaaacatatt tcgatcaatt aaaccaggat 6901 tcataaaaat ataagtatat atataaatga tgtttcgttt agcgatgaac ttcactcata 6961 tgataatact taacaatata agtacataaa aaataaaata aaattaattg tttacgaaaa 7021 gtctacaaat actgcatgta taattaatgt tctctttatt tatttattta taccttacca 7081 agatatatct ataaccgcat agaaatagaa ggcgaagaga taatttccaa aaacaagaaa 7141 aacctctaag ctcaaaaggg ccggccatga ttgaacaaga tggattgcac gcaggttctc 7201 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 7261 ctgatgccgc cgtgttccgg ctgtcagcgc aggggaggcc ggttcttttt gtcaagaccg 7321 acctgtccgg tgccctgaat gaacttcaag acgaggcagc gcggctatcg tggctggcca 7381 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 7441 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 7501 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 7561 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 7621 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 7681 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgactcat ggcgatgcct 7741 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 7801 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 7861 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 7921 agcgcatcgc cttctatcgc cttcttgacg agttcttctg aggcgcgcc //