LOCUS       Exported                9393 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pLH6500, complete sequence.
ACCESSION   AY234329
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9393)
  AUTHORS   Hausmann L, Toepfer R.
  TITLE     Development of Plasmid Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9393)
  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 9393)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9393
                     /organism="Binary vector pLH6500"
                     /mol_type="other DNA"
                     /db_xref="taxon:226214"
     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            438..1492
                     /gene="hph"
                     /gene_synonym="hpt; hyg"
                     /label=hph
                     /note="hygromycin resistance gene from Escherichia coli; 
                     similar to GenBank Accession Number K01193"
     misc_difference 447
                     /gene="hph"
                     /gene_synonym="hpt; hyg"
                     /replace="g"
                     /label=compared to GenBank accesion Number K01193
                     /note="compared to GenBank accesion Number K01193"
     CDS             452..1477
                     /codon_start=1
                     /gene="hph"
                     /gene_synonym="hpt; hyg"
                     /product="hygromycin phosphotransferase"
                     /label=hph
                     /note="confers resistance to hygromycin"
                     /protein_id="AAO85355.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      1503..1702
                     /regulatory_class="terminator"
                     /note="terminator sequence from the 35S gene of Cauliflower
                     mosaic virus in GenBank Accession Numbers X05868 and 
                     V00140"
     misc_difference 1541
                     /label=compared to GenBank Accession Number X05868
                     /note="compared to GenBank Accession Number X05868"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      1668..1683
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    1698
                     /label=putative transcription stop site
                     /note="putative transcription stop site"
     misc_feature    1721..1822
                     /note="multiple cloning site from vector pBlueSfi AB in 
                     GenBank Accession Number AF327874"
     regulatory      1792..1801
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1823..2425
                     /note="right border region from nopaline-type Ti plasmid 
                     C58 of Agrobacterium tumefaciens in GenBank Accession 
                     Number X07435"
     misc_difference 1848
                     /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    1867..1891
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1868..1891
                     /label=right flanking T-DNA border
                     /note="right flanking T-DNA border"
     repeat_region   1915..1927
                     /label=repeat C
                     /note="repeat C"
     misc_difference 1921
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   1950..1961
                     /label=repeat C
                     /note="repeat C"
     repeat_region   1975..1985
                     /label=repeat B
                     /note="repeat B"
     repeat_region   2002..2012
                     /label=repeat B
                     /note="repeat B"
     misc_difference 2007
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 2018
                     /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(2035..2425)
                     /note="3 sequence of the ornithine cyclodeaminase gene 
                     ocd; non-functional"
     misc_difference 2211
                     /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      2426..3590
                     /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      2536..3124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_difference 2683
                     /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 3222..3223
                     /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    3309..3449
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    3402
                     /label=nic/bom site
                     /note="nic/bom site"
     rep_origin      3606..7377
                     /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      3737..3742
                     /regulatory_class="minus_35_signal"
     regulatory      3763..3768
                     /regulatory_class="minus_10_signal"
     regulatory      3785..3789
                     /regulatory_class="ribosome_binding_site"
     CDS             3799..4485
                     /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="AAO85356.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     misc_difference 4218
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     stem_loop       4629..4661
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_difference 4637
                     /replace="g"
                     /note="nucleotide changed to conserve palindromic sequence 
                     following SfiI restriction site-destruction"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 4652
                     /replace="c"
                     /note="original nucleotide changed to destroy two SfiI 
                     restriction sites"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      4719..4724
                     /regulatory_class="minus_35_signal"
     regulatory      4740..4745
                     /regulatory_class="minus_10_signal"
     regulatory      4773..4780
                     /regulatory_class="ribosome_binding_site"
     CDS             4784..5413
                     /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="AAO85357.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5680..5692
                     /note="identical to the korB operator binding site of 
                     plasmid RK2 in GenBank Accession Number L27758.1"
     CDS             5740..6915
                     /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="AAO85358.1"
                     /translation="MPHVWRNGRLARRKRLGCLPALQWHWNPQARGIGVSGRKPSGPVQ
                     IGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKP
                     PKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGP
                     AGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKD
                     DLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELAD
                     WMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSA
                     GEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             5842..6915
                     /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 5846
                     /label=compared to GenBank Accession Number U10487
                     /note="compared to GenBank Accession Number U10487"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 5919
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 6240
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   6907..6921
                     /label=repeat D
                     /note="repeat D"
     stem_loop       6929..6951
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_feature    6958..7377
                     /note="parC region; contains putative cis-acting sites for 
                     replication, partition and incompatibility"
     rep_origin      6981..7175
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     protein_bind    6981..6989
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   6992..7006
                     /label=repeat D
                     /note="repeat D"
     repeat_region   7013..7027
                     /label=repeat D
                     /note="repeat D"
     repeat_region   7034..7048
                     /label=repeat D
                     /note="repeat D"
     repeat_region   7055..7069
                     /label=repeat D
                     /note="repeat D"
     misc_feature    7078..7156
                     /label=AT-rich region
                     /note="AT-rich region"
     protein_bind    7168..7176
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   7272..7285
                     /label=repeat E
                     /rpt_unit_range=7272 .. 7278
                     /note="repeat E"
     misc_feature    7385..8789
                     /label=similar to GenBank Accession Number M35007
                     /note="similar to GenBank Accession Number M35007"
     regulatory      7516..7521
                     /regulatory_class="minus_35_signal"
     regulatory      7539..7544
                     /regulatory_class="minus_10_signal"
     CDS             7561..8571
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /label=aadA
                     /note="confers resistance to streptomycin and 
                     spectinomycin"
                     /protein_id="AAO85359.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            7561..8571
                     /gene="aadA"
                     /label=aadA
                     /note="different start codons are described in the 
                     literature"
     CDS             7780..8571
                     /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       8574..8631
                     /note="palindromic sequence; putative termination signal"
     misc_feature    8790..9391
                     /note="left border region from octopine-type Ti plasmid of 
                     Agrobacterium tumefaciens; similar to GenBank Accession 
                     Number X00493"
     misc_difference 8808..8809
                     /replace="ggtacc"
                     /note="compared to GenBank Accession Number X00493; removed
                     to destroy a KpnI restriction site"
     misc_feature    9091..9115
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    9092..9115
                     /label=left flanking T-DNA border
                     /note="left flanking T-DNA border"