LOCUS       Exported                9401 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pLH6000, complete sequence.
ACCESSION   AY234328
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9401)
  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 9401)
  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 9401)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9401
                     /organism="Binary vector pLH6000"
                     /mol_type="other DNA"
                     /db_xref="taxon:226213"
     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="AAO85350.1"
                     /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    1729..1830
                     /note="multiple cloning site from vector pBlueSfi BA in 
                     GenBank Accession Number AF327875"
     regulatory      1736..1745
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1831..2433
                     /note="right border region from nopaline-type Ti plasmid 
                     C58 of Agrobacterium tumefaciens in GenBank Accession 
                     Number X07435"
     misc_difference 1856
                     /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    1875..1899
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1876..1899
                     /label=right flanking T-DNA border
                     /note="right flanking T-DNA border"
     repeat_region   1923..1935
                     /label=repeat C
                     /note="repeat C"
     misc_difference 1929
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   1958..1969
                     /label=repeat C
                     /note="repeat C"
     repeat_region   1983..1993
                     /label=repeat B
                     /note="repeat B"
     repeat_region   2010..2020
                     /label=repeat B
                     /note="repeat B"
     misc_difference 2015
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 2026
                     /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(2043..2433)
                     /note="3 sequence of the ornithine cyclodeaminase gene 
                     ocd; non-functional"
     misc_difference 2219
                     /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      2434..3598
                     /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      2544..3132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_difference 2691
                     /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 3230..3231
                     /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    3317..3457
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    3410
                     /label=nic/bom site
                     /note="nic/bom site"
     rep_origin      3614..7385
                     /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      3745..3750
                     /regulatory_class="minus_35_signal"
     regulatory      3771..3776
                     /regulatory_class="minus_10_signal"
     regulatory      3793..3797
                     /regulatory_class="ribosome_binding_site"
     CDS             3807..4493
                     /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="AAO85351.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     misc_difference 4226
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     stem_loop       4637..4669
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_difference 4645
                     /replace="g"
                     /note="nucleotide changed to conserve palindromic sequence 
                     following SfiI restriction site-destruction"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 4660
                     /replace="c"
                     /note="original nucleotide changed to destroy two SfiI 
                     restriction sites"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      4727..4732
                     /regulatory_class="minus_35_signal"
     regulatory      4748..4753
                     /regulatory_class="minus_10_signal"
     regulatory      4781..4788
                     /regulatory_class="ribosome_binding_site"
     CDS             4792..5421
                     /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="AAO85352.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5688..5700
                     /note="identical to the korB operator binding site of 
                     plasmid RK2 in GenBank Accession Number L27758.1"
     CDS             5748..6923
                     /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="AAO85353.1"
                     /translation="MPHVWRNGRLARRKRLGCLPALQWHWNPQARGIGVSGRKPSGPVQ
                     IGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKP
                     PKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGP
                     AGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKD
                     DLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELAD
                     WMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSA
                     GEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             5850..6923
                     /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 5854
                     /label=compared to GenBank Accession Number U10487
                     /note="compared to GenBank Accession Number U10487"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 5927
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 6248
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   6915..6929
                     /label=repeat D
                     /note="repeat D"
     stem_loop       6937..6959
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_feature    6966..7385
                     /note="parC region; contains putative cis-acting sites for 
                     replication, partition and incompatibility"
     rep_origin      6989..7183
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     protein_bind    6989..6997
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   7000..7014
                     /label=repeat D
                     /note="repeat D"
     repeat_region   7021..7035
                     /label=repeat D
                     /note="repeat D"
     repeat_region   7042..7056
                     /label=repeat D
                     /note="repeat D"
     repeat_region   7063..7077
                     /label=repeat D
                     /note="repeat D"
     misc_feature    7086..7164
                     /label=AT-rich region
                     /note="AT-rich region"
     protein_bind    7176..7184
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   7280..7293
                     /label=repeat E
                     /rpt_unit_range=7280 .. 7286
                     /note="repeat E"
     misc_feature    7393..8797
                     /label=similar to GenBank Accession Number M35007
                     /note="similar to GenBank Accession Number M35007"
     regulatory      7524..7529
                     /regulatory_class="minus_35_signal"
     regulatory      7547..7552
                     /regulatory_class="minus_10_signal"
     CDS             7569..8579
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /label=aadA
                     /note="confers resistance to streptomycin and 
                     spectinomycin"
                     /protein_id="AAO85354.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            7569..8579
                     /gene="aadA"
                     /label=aadA
                     /note="different start codons are described in the 
                     literature"
     CDS             7788..8579
                     /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       8582..8639
                     /note="palindromic sequence; putative termination signal"
     misc_feature    8798..9399
                     /note="left border region from octopine-type Ti plasmid of 
                     Agrobacterium tumefaciens; similar to GenBank Accession 
                     Number X00493"
     misc_difference 8816..8817
                     /replace="ggtacc"
                     /note="compared to GenBank Accession Number X00493; removed
                     to destroy a KpnI restriction site"
     misc_feature    9099..9123
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    9100..9123
                     /label=left flanking T-DNA border
                     /note="left flanking T-DNA border"