LOCUS V12_with_biobrick 8203 bp ds-DNA circular 18-JUN-2010 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and Hegedus,D.D. TITLE pORE: a new series of binary vectors with a removable selectable marker and 21 site MCS JOURNAL Unpublished REFERENCE 2 (bases 1 to 8255) AUTHORS Coutu,C.M., Brandle,J., Brown,K., Simmonds,J., Brown,D., Mici,B. and Hegedus,D.D. TITLE Direct Submission JOURNAL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 COMMENT COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers source join(1..4760,5553..8203) /organism="Binary vector pORE_R3" /mol_type="other DNA" /db_xref="taxon:267423" /lab_host="Agrobacterium tumefaciens" /label=source:Binary vector pORE_R3 /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=pink /ApEinfo_revcolor=pink misc_feature 356..516 /note="T-DNA left border" /label=T-DNA left border /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink 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="AAS78971.1" /db_xref="GI:45862392" /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="AAS78972.1" /db_xref="GI:45862393" /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=pink /ApEinfo_revcolor=pink 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; selected unique sites: SfoI-SacII-HindIII-NheI-MfeI-PspOMI-AclI-XhoI-XbaI-StuI- BamHI-SmaI-EcoRI-SalI-NotI-ClaI-PstI-SacI" /label=multiple cloning site; selected unique sites: SfoI /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink primer_bind complement(5564..5586) /note="M13/pUC reverse" /PCR_conditions="60 degree C annealing temperature" /label=M13/pUC reverse /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink terminator 5593..5845 /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 5852..5899 /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 5916..7392 /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=pink /ApEinfo_revcolor=pink CDS 7401..8195 /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" /label=NptII /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink misc_feature 4766..4814 /label=biobrick cloning site /ApEinfo_fwdcolor=#14ff00 /ApEinfo_revcolor=green gene 4822..5552 /label=smgfp reporter gene /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green primer_bind 4588..4607 /label=sequencing forward primer binding site /ApEinfo_fwdcolor=#efff00 /ApEinfo_revcolor=green misc_feature 5092..5111 /label=squencing forward primer binding site /ApEinfo_fwdcolor=#fffe00 /ApEinfo_revcolor=green misc_feature 4909..4928 /label=sequencing reverse primer binding site /ApEinfo_fwdcolor=#f9ff00 /ApEinfo_revcolor=green misc_feature 5427..5446 /label=sequencing reverse primer binding site(1) /ApEinfo_label=sequencing reverse primer binding site /ApEinfo_fwdcolor=#fff700 /ApEinfo_revcolor=green misc_feature 5926..5945 /label=sequencing reverse primer binding site(2) /ApEinfo_label=sequencing reverse primer binding site /ApEinfo_fwdcolor=#edff00 /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 AGCTTGAATT CGCGGCCGCT TCTAGAATTA GAGGACTAGT 4801 AGCGGCCGCT GCAGAGAGCT CATGGCGAGT AAAGGAGAAG AACTTTTCAC TGGAGTTGTC 4861 CCAATTCTTG TTGAATTAGA TGGTGATGTT AATGGGCACA AATTTTCTGT CAGTGGAGAG 4921 GGTGAAGGTG ATGCAACATA CGGAAAACTT ACCCTTAAAT TTATTTGCAC TACTGGAAAA 4981 CTACCTGTTC CTTGGCCAAC ACTTGTCACT ACTTTCTCTT ATGGTGTTCA ATGCTTTTCA 5041 AGATACCCAG ATCATATGAA GCGGCACGAC TTCTTCAAGA GCGCCATGCC TGAGGGATAC 5101 GTGCAGGAGA GGACCATCTC TTTCAAGGAC GACGGGAACT ACAAGACACG TGCTGAAGTC 5161 AAGTTTGAGG GAGACACCCT CGTCAACAGG ATCGAGCTTA AGGGAATTGA TTTCAAGGAG 5221 GACGGAAACA TCCTCGGCCA CAAGTTGGAA TACAACTACA ACTCCCACAA CGTATACATC 5281 ACGGCAGACA AACAAAAGAA TGGAATCAAA GCTAACTTCA AAATTAGACA CAACATTGAA 5341 GATGGAAGCG TTCAACTAGC AGACCATTAT CAACAAAATA CTCCTATTGG CGATGGCCCT 5401 GTCCTTTTAC CAGACAACCA TTACCTGTCC ACACAATCTG CCCTTTCGAA AGATCCCAAC 5461 GAAAAGAGAG ACCACATGGT CCTTCTTGAG TTTGTAACAG CTGCTGGGAT TACACATGGC 5521 ATGGATGAAC TATACAAATA AGGTACCAAA AGctagtgat atccctgtgt gaaattgtta 5581 tccgctacgc gtgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt 5641 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt 5701 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta 5761 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc 5821 gcggtgtcat ctatgttact agatcccatg ggaagttcct attccgaagt tcctattctc 5881 tgaaaagtat aggaacttca gcgatcgcag acgtcaacgt ggatacttgg cagtggttac 5941 ttggcttttc ctttattttc ttttggacgg aagcggtggt tactttgtca cacatttaaa 6001 aaaacacgtg tttctcactt ttttctattc ccgtcacaaa caattttaag aaagatccat 6061 ctatcgtgat ctttctatca aacaaaagaa aaaaggtctt catagtaacg ctacaacatc 6121 aaatatgtgg ttgctctgac atcagtcggg aaaataagga tatggcggca ttggccacat 6181 ctattggggt cccaacttcc tttcacaaaa aaattaaatt gggtgtccca acttttatct 6241 ttgatatagt gacatgagta tcgggagcat tggacaatgg ataaaatgag aactaaaaaa 6301 attctggtta atttttgatc attgttattt aaaaggttat tttatctata atctacccat 6361 attgatcagt tttatttaaa tttgtttagc taccgctcca cgagagagat cctcatctta 6421 aaaatggaat atggaaatta cacacgaccc caaaagtata ttttttctct ggagaatgct 6481 atttagagct ttgactatat ggtctgaatt agaaagacgg gaaataaaat ctgctaagtg 6541 atataagctc taagtaggcg atgtgtgatg gagaacacct tttctttaac agtcttcatg 6601 ttttacagat tcgcgaactt cgaatatccc tatacggtct gtctaaccct cgtgtgtctt 6661 ttgagtccaa gataaaggcc attattgagt aacatagaca tgctggaatc caaccattga 6721 agtcacaact gtccatgtag attctttgga gaatctgaaa agtcttaata aaggtggtgt 6781 ttcaaagaaa acaaaacaaa tgagttaaga aaaaaaaata tcatgtagtg gtcgagtatt 6841 atgttattta ttgtgtagct accaatcttt attctttaaa tctgacataa aatgctacaa 6901 actttttacc tcgtctatag ccccaaaaaa cctaaccacg gttctaaaac cacacacagt 6961 gattttggtt gacgacaatg cctctccttc ctcaaaacga tttatttaca ttttttaaat 7021 caaatgttac attttatacc ataattaagt ctttttacag aatacttaga tggaagagat 7081 gtataaaaaa ggaggaaatt gtaaaaaaca tatttcgatc aattaaacca ggattcataa 7141 aaatataagt atatatataa atgatgtttc gtttagcgat gaacttcact catatgataa 7201 tacttaacaa tataagtaca taaaaaataa aataaaatta attgtttacg aaaagtctac 7261 aaatactgca tgtataatta atgttctctt tatttattta tttatacctt accaagatat 7321 atctataacc gcatagaaat agaaggcgaa gagataattt ccaaaaacaa gaaaaacctc 7381 taagctcaaa agggccggcc atgattgaac aagatggatt gcacgcaggt tctccggccg 7441 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 7501 ccgccgtgtt ccggctgtca gcgcagggga ggccggttct ttttgtcaag accgacctgt 7561 ccggtgccct gaatgaactt caagacgagg cagcgcggct atcgtggctg gccacgacgg 7621 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 7681 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 7741 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 7801 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 7861 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 7921 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac tcatggcgat gcctgcttgc 7981 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 8041 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 8101 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 8161 tcgccttcta tcgccttctt gacgagttct tctgaggcgc gcc //