LOCUS       Exported                8958 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pLH5000, complete sequence.
ACCESSION   AY234327
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8958)
  AUTHORS   Hausmann L, Toepfer R.
  TITLE     Development of Plasmid Vectors
  JOURNAL   (in) Brauer,D., Roebbelen,G. and Toepfer,R. (Eds.); BIOENGINEERING 
            OF CUSTOM-TAILORED RAPE VARIETIES: 155-172; GPZ e. V., Von 
            Sieboldstr. 8, Goettingen, Germany (1999)
REFERENCE   2  (bases 1 to 8958)
  AUTHORS   Hausmann L, Toepfer R.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2003) Institute for Grapevine Breeding 
            Geilweilerhof, Siebeldingen 76833, Germany
REFERENCE   3  (bases 1 to 8958)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8958
                     /organism="Binary vector pLH5000"
                     /mol_type="other DNA"
                     /db_xref="taxon:226212"
     regulatory      6..424
                     /regulatory_class="promoter"
                     /note="promoter/enhancer sequence from the 35S gene of 
                     Cauliflower mosaic virus in GenBank Accession Numbers 
                     X05868 and V00140"
     promoter        80..424
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      360..363
                     /regulatory_class="CAAT_signal"
     regulatory      392..398
                     /regulatory_class="TATA_box"
     misc_feature    423
                     /label=putative transcription start site
                     /note="putative transcription start site"
     gene            439..1051
                     /gene="dhfr"
                     /gene_synonym="mtx"
                     /label=dhfr
                     /note="methotrexate resistance gene from Mus musculus; 
                     similar to GenBank Accession Number V00734"
     CDS             446..1009
                     /codon_start=1
                     /gene="dhfr"
                     /gene_synonym="mtx"
                     /product="dihydrofolate reductase"
                     /label=dhfr
                     /note="confers resistance to methotrexate"
                     /protein_id="AAO85345.1"
                     /translation="MVRPLNCIVAVSQNMGIGKNGDRPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     regulatory      1060..1259
                     /regulatory_class="terminator"
                     /note="terminator sequence from the 35S gene of Cauliflower
                     mosaic virus in GenBank Accession Numbers X05868 and 
                     V00140"
     misc_difference 1098
                     /label=compared to GenBank Accession Number X05868
                     /note="compared to GenBank Accession Number X05868"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      1225..1240
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    1255
                     /label=putative transcription stop site
                     /note="putative transcription stop site"
     misc_feature    1286..1387
                     /note="multiple cloning site from vector pBlueSfi BA in 
                     GenBank Accession Number AF327875"
     regulatory      1293..1302
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1388..1990
                     /note="right border region from nopaline-type Ti plasmid 
                     C58 of Agrobacterium tumefaciens in GenBank Accession 
                     Number X07435"
     misc_difference 1413
                     /replace="a"
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_feature    1432..1456
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1433..1456
                     /label=right flanking T-DNA border
                     /note="right flanking T-DNA border"
     repeat_region   1480..1492
                     /label=repeat C
                     /note="repeat C"
     misc_difference 1486
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   1515..1526
                     /label=repeat C
                     /note="repeat C"
     repeat_region   1540..1550
                     /label=repeat B
                     /note="repeat B"
     repeat_region   1567..1577
                     /label=repeat B
                     /note="repeat B"
     misc_difference 1572
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 1583
                     /replace="a"
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_feature    complement(1600..1990)
                     /note="3 sequence of the ornithine cyclodeaminase gene 
                     ocd; non-functional"
     misc_difference 1776
                     /replace="t"
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     rep_origin      1991..3155
                     /note="ColE1 origin of replication sequence of pBR322; 
                     similar to GenBank Accession Number J01749; serves as 
                     origin of replication in Escherichia coli host"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     rep_origin      2101..2689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_difference 2248
                     /replace="c"
                     /label=compared to GenBank Accession Number J01749
                     /note="compared to GenBank Accession Number J01749"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 2787..2788
                     /replace="gta"
                     /label=compared to GenBank Accession Number J01749
                     /note="compared to GenBank Accession Number J01749"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_feature    2874..3014
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    2967
                     /label=nic/bom site
                     /note="nic/bom site"
     rep_origin      3171..6942
                     /note="origin of replication and partitioning region from 
                     plasmid pVS1 of Pseudomonas; similar to GenBank Accession 
                     Number U10487; serves as origin of replication in 
                     Agrobacterium tumefaciens host"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      3302..3307
                     /regulatory_class="minus_35_signal"
     regulatory      3328..3333
                     /regulatory_class="minus_10_signal"
     regulatory      3350..3354
                     /regulatory_class="ribosome_binding_site"
     CDS             3364..4050
                     /codon_start=1
                     /product="resolvase"
                     /label=resolvase
                     /note="resolvase-like protein encoded by parR of plasmid 
                     pVS1; similar to Tn3 resolvase in GenBank Accession Number 
                     V00613, Tn917 resolvase in GenBank Accession Number M11180,
                     and RK2 ParA in GenBank Accession Number L27758"
                     /protein_id="AAO85346.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     misc_difference 3783
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     stem_loop       4194..4226
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_difference 4202
                     /replace="g"
                     /note="nucleotide changed to conserve palindromic sequence 
                     following SfiI restriction site-destruction"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 4217
                     /replace="c"
                     /note="original nucleotide changed to destroy two SfiI 
                     restriction sites"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      4284..4289
                     /regulatory_class="minus_35_signal"
     regulatory      4305..4310
                     /regulatory_class="minus_10_signal"
     regulatory      4338..4345
                     /regulatory_class="ribosome_binding_site"
     CDS             4349..4978
                     /codon_start=1
                     /product="ParA"
                     /label=ParA
                     /note="partitioning protein ParA encoded by parA of plasmid
                     pVS1; similar to F plasmid SopA in GenBank Accession Number
                     X04619, RK2 IncC in GenBank Accession Number L27758, and P1
                     ParA in GenBank Accession Number X02954"
                     /protein_id="AAO85347.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5245..5257
                     /note="identical to the korB operator binding site of 
                     plasmid RK2 in GenBank Accession Number L27758.1"
     CDS             5305..6480
                     /codon_start=1
                     /product="ParB"
                     /label=ParB
                     /note="partitioning protein ParB encoded by parB of plasmid
                     pVS1; similar to F plasmid SopB in GenBank Accession Number
                     X04619, RK2 KorB in GenBank Accession Number L27758, and P1
                     ParB in GenBank Accession Number X02954"
                     /protein_id="AAO85348.1"
                     /translation="MPHVWRNGRLARRKRLGCLPALQWHWNPQARGIGVSGRKPSGPVQ
                     IGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKP
                     PKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGP
                     AGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKD
                     DLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELAD
                     WMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSA
                     GEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             5407..6480
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     misc_difference 5411
                     /label=compared to GenBank Accession Number U10487
                     /note="compared to GenBank Accession Number U10487"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 5484
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 5805
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   6472..6486
                     /label=repeat D
                     /note="repeat D"
     stem_loop       6494..6516
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_feature    6523..6942
                     /note="parC region; contains putative cis-acting sites for 
                     replication, partition and incompatibility"
     rep_origin      6546..6740
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     protein_bind    6546..6554
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   6557..6571
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6578..6592
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6599..6613
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6620..6634
                     /label=repeat D
                     /note="repeat D"
     misc_feature    6643..6721
                     /label=AT-rich region
                     /note="AT-rich region"
     protein_bind    6733..6741
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   6837..6850
                     /label=repeat E
                     /rpt_unit_range=6837 .. 6843
                     /note="repeat E"
     misc_feature    6950..8354
                     /label=similar to GenBank Accession Number M35007
                     /note="similar to GenBank Accession Number M35007"
     regulatory      7081..7086
                     /regulatory_class="minus_35_signal"
     regulatory      7104..7109
                     /regulatory_class="minus_10_signal"
     CDS             7126..8136
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /label=aadA
                     /note="confers resistance to streptomycin and 
                     spectinomycin"
                     /protein_id="AAO85349.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            7126..8136
                     /gene="aadA"
                     /label=aadA
                     /note="different start codons are described in the 
                     literature"
     CDS             7345..8136
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     stem_loop       8139..8196
                     /note="palindromic sequence; putative termination signal"
     misc_feature    8355..8956
                     /note="left border region from octopine-type Ti plasmid of 
                     Agrobacterium tumefaciens; similar to GenBank Accession 
                     Number X00493"
     misc_difference 8373..8374
                     /replace="ggtacc"
                     /note="compared to GenBank Accession Number X00493; removed
                     to destroy a KpnI restriction site"
     misc_feature    8656..8680
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    8657..8680
                     /label=left flanking T-DNA border
                     /note="left flanking T-DNA border"