LOCUS       Exported                9146 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pLH9500, complete sequence.
ACCESSION   AF458479
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9146)
  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; Gesellschaft fuer 
            Pflanzenzuechtung e. V., Von Sieboldstr. 8, Goettingen, Germany 
            (1999)
REFERENCE   2  (bases 1 to 9146)
  AUTHORS   Hausmann L, Toepfer R.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2001) Institute for Grapevine Breeding 
            Geilweilerhof, Siebeldingen 76833, Germany
REFERENCE   3  (bases 1 to 9146)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9146
                     /organism="Binary vector pLH9500"
                     /plasmid="pLH9500"
                     /mol_type="other DNA"
                     /db_xref="taxon:182805"
     misc_feature    6..424
                     /note="promoter/enhancer sequence from the 35S gene of 
                     Cauliflower mosaic virus: GenBank Accession Number X05868, 
                     M31331"
     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"
     regulatory      433..442
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     gene            439..1249
                     /gene="nptII"
                     /label=nptII
                     /note="neomycin/kanamycin resistance gene of Tn5 from 
                     Escherichia coli; similar to GenBank Accession Number 
                     U00004"
     CDS             439..1242
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /note="NPTII; confers resistance to kanamycin and neomycin"
                     /protein_id="AAL78963.1"
                     /translation="MGAWIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             451..1242
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      1256..1455
                     /regulatory_class="terminator"
                     /note="terminator sequence from the 35S gene of Cauliflower
                     mosaic virus; GenBank Accession Number: X05868, M31331"
     misc_difference 1289
                     /label=compared to GenBank Accession Number X05868
                     /note="compared to GenBank Accession Number X05868"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      1421..1436
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    1451
                     /label=putative transcription stop site
                     /note="putative transcription stop site"
     misc_feature    1474..1575
                     /note="multiple cloning site from vector pBlueSfi AB; 
                     GenBank Accession Number: AF327874"
                     /citation="[1]"
     regulatory      1545..1554
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1576..2178
                     /note="right border region from nopaline-type Ti plasmid 
                     C58 of Agrobacterium tumefaciens; GenBank Accession Number:
                     X07435"
     misc_difference 1601
                     /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    1620..1644
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1621..1644
                     /label=right flanking T-DNA border
                     /note="right flanking T-DNA border"
     repeat_region   1668..1680
                     /label=repeat C
                     /note="repeat C"
     misc_difference 1674
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   1703..1714
                     /label=repeat C
                     /note="repeat C"
     repeat_region   1728..1738
                     /label=repeat B
                     /note="repeat B"
     repeat_region   1755..1765
                     /label=repeat B
                     /note="repeat B"
     misc_difference 1760
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 1771
                     /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    1788..2178
                     /note="3 sequence of the ornithine cyclodeaminase gene ocd
                     (non-functional)"
     misc_difference 1964
                     /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      2179..3343
                     /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      2289..2877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_difference 2436
                     /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 2975..2976
                     /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    3062..3202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    3155
                     /label=nic/bom site
                     /note="nic/bom site"
     rep_origin      3359..7130
                     /note="origin of replication and partitioning region from 
                     plasmid pVS1 of Pseudomonas (Itoh et al. 1984, Plasmid 11: 
                     206-220); similar to GenBank Accession Number U10487; 
                     serves as origin of replication in Agrobacterium 
                     tumefaciens host"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      3490..3495
                     /regulatory_class="minus_35_signal"
     regulatory      3516..3521
                     /regulatory_class="minus_10_signal"
     regulatory      3538..3542
                     /regulatory_class="ribosome_binding_site"
     CDS             3552..4238
                     /codon_start=1
                     /product="resolvase"
                     /label=resolvase
                     /note="resolvase-like protein encoded by parR of plasmid 
                     pVS1; similar to Tn3 resolvase (GenBank Accession Number 
                     V00613), Tn917 resolvase (GenBank Accession Number M11180) 
                     and ParA (RK2) (GenBank Accession Number L27758)"
                     /protein_id="AAL78966.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     misc_difference 3971
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     stem_loop       4382..4414
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_difference 4390
                     /replace="g"
                     /note="nucleotide changed to conserve palindromic sequence 
                     following SfiI restriction site-destruction"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 4405
                     /replace="c"
                     /note="original nucleotide changed to destroy two SfiI 
                     restriction sites"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      4472..4477
                     /regulatory_class="minus_35_signal"
     regulatory      4493..4498
                     /regulatory_class="minus_10_signal"
     regulatory      4526..4533
                     /regulatory_class="ribosome_binding_site"
     CDS             4537..5166
                     /codon_start=1
                     /product="ParA"
                     /label=ParA
                     /note="partitioning protein ParA encoded by parA of plasmid
                     pVS1; similar to SopA (F plasmid) (GenBank Accession Number
                     X04619), IncC (RK2) (GenBank Accession Number L27758), and 
                     ParA (P1) (GenBank Accession Number X02954)"
                     /protein_id="AAL78964.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5433..5445
                     /note="identical to the korB operator binding site of 
                     plasmid RK2 (Pansegrau et al. 1994, J. Mol. Biol. 239: 
                     623-663); GenBank Accession Number L27758"
     CDS             5493..6668
                     /codon_start=1
                     /product="ParB"
                     /label=ParB
                     /note="partitioning protein ParB encoded by parB of plasmid
                     pVS1; similar to SopB (F plasmid) (GenBank Accession Number
                     X04619), KorB (RK2) (GenBank Accession Number L27758), and 
                     ParB (P1) (GenBank Accession Number X02954)"
                     /protein_id="AAL78965.1"
                     /translation="MPHVWRNGRLARRKRLGCLPALQWHWNPQARGIGVSGRKPSGPVQ
                     IGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKP
                     PKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGP
                     AGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKD
                     DLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELAD
                     WMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSA
                     GEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             5595..6668
                     /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 5599
                     /label=compared to GenBank Accession Number U10487
                     /note="compared to GenBank Accession Number U10487"
     misc_difference 5672
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 5993
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   6660..6674
                     /label=repeat D
                     /note="repeat D"
     stem_loop       6682..6704
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_feature    6711..7130
                     /note="parC region; contains putative cis-acting sites for 
                     replication, partition and incompatibility"
     rep_origin      6734..6928
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     protein_bind    complement(6734..6742)
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   6745..6759
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6766..6780
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6787..6801
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6808..6822
                     /label=repeat D
                     /note="repeat D"
     misc_feature    6831..6909
                     /label=AT-rich region
                     /note="AT-rich region"
     protein_bind    complement(6921..6929)
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   7025..7031
                     /label=repeat E
                     /note="repeat E"
     repeat_region   7032..7038
                     /label=repeat E
                     /note="repeat E"
     misc_feature    7138..8542
                     /label=similar to GenBank Accession Number M35007
                     /note="similar to GenBank Accession Number M35007"
     regulatory      7269..7274
                     /regulatory_class="minus_35_signal"
     regulatory      7292..7297
                     /regulatory_class="minus_10_signal"
     CDS             7314..8324
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /function="confers resistance to streptomycin and 
                     spectinomycin"
                     /label=aadA
                     /note="different start codons are described in the 
                     literature"
                     /protein_id="AAL78962.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            7314..8324
                     /gene="aadA"
                     /label=aadA
     CDS             7533..8324
                     /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       8327..8384
                     /note="palindromic sequence; putative termination signal"
     misc_feature    8543..9144
                     /note="left border region from octopine-type Ti plasmid of 
                     Agrobacterium tumefaciens; similar to GenBank Accession 
                     Number X00493"
     misc_difference 8561..8562
                     /replace="ggtacc"
                     /note="compared to GenBank Accession Number X00493; removed
                     to destroy a KpnI restriction site"
     misc_feature    8844..8868
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    8845..8868
                     /label=left flanking T-DNA border
                     /note="left flanking T-DNA border"