LOCUS       Exported                9154 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pLH9000, complete sequence.
ACCESSION   AF458478
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9154)
  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 9154)
  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 9154)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9154
                     /organism="Binary vector pLH9000"
                     /plasmid="pLH9000"
                     /mol_type="other DNA"
                     /db_xref="taxon:182804"
     regulatory      6..424
                     /regulatory_class="promoter"
                     /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="AAL78958.1"
                     /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    1482..1583
                     /note="multiple cloning site from vector pBlueSfi BA; 
                     GenBank Accession Number: AF327875"
                     /citation="[1]"
     regulatory      1489..1498
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1584..2186
                     /note="right border region from nopaline-type Ti plasmid 
                     C58 of Agrobacterium tumefaciens; GenBank Accession Number:
                     X07435"
     misc_difference 1609
                     /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    1628..1652
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1629..1652
                     /label=right flanking T-DNA border
                     /note="right flanking T-DNA border"
     repeat_region   1676..1688
                     /label=repeat C
                     /note="repeat C"
     misc_difference 1682
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   1711..1722
                     /label=repeat C
                     /note="repeat C"
     repeat_region   1736..1746
                     /label=repeat B
                     /note="repeat B"
     repeat_region   1763..1773
                     /label=repeat B
                     /note="repeat B"
     misc_difference 1768
                     /label=compared to GenBank Accession Number X07435
                     /note="compared to GenBank Accession Number X07435"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 1779
                     /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    1796..2186
                     /note="3 sequence of the ornithine cyclodeaminase gene ocd
                     (non-functional)"
     misc_difference 1972
                     /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      2187..3351
                     /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      2297..2885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_difference 2444
                     /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 2983..2984
                     /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    3070..3210
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    3163
                     /label=nic/bom site
                     /note="nic/bom site"
     rep_origin      3367..7138
                     /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      3498..3503
                     /regulatory_class="minus_35_signal"
     regulatory      3524..3529
                     /regulatory_class="minus_10_signal"
     regulatory      3546..3550
                     /regulatory_class="ribosome_binding_site"
     CDS             3560..4246
                     /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="AAL78961.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     misc_difference 3979
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     stem_loop       4390..4422
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_difference 4398
                     /replace="g"
                     /note="nucleotide changed to conserve palindromic sequence 
                     following SfiI restriction site-destruction"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 4413
                     /replace="c"
                     /note="original nucleotide changed to destroy two SfiI 
                     restriction sites"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      4480..4485
                     /regulatory_class="minus_35_signal"
     regulatory      4501..4506
                     /regulatory_class="minus_10_signal"
     regulatory      4534..4541
                     /regulatory_class="ribosome_binding_site"
     CDS             4545..5174
                     /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="AAL78959.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5441..5453
                     /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             5501..6676
                     /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="AAL78960.1"
                     /translation="MPHVWRNGRLARRKRLGCLPALQWHWNPQARGIGVSGRKPSGPVQ
                     IGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKP
                     PKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGP
                     AGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKD
                     DLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELAD
                     WMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSA
                     GEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             5603..6676
                     /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 5607
                     /label=compared to GenBank Accession Number U10487
                     /note="compared to GenBank Accession Number U10487"
     misc_difference 5680
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 6001
                     /replace="c"
                     /note="original nucleotide changed to destroy a SfiI 
                     restriction site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     repeat_region   6668..6682
                     /label=repeat D
                     /note="repeat D"
     stem_loop       6690..6712
                     /label=palindromic sequence
                     /note="palindromic sequence"
     misc_feature    6719..7138
                     /note="parC region; contains putative cis-acting sites for 
                     replication, partition and incompatibility"
     rep_origin      6742..6936
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     protein_bind    complement(6742..6750)
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   6753..6767
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6774..6788
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6795..6809
                     /label=repeat D
                     /note="repeat D"
     repeat_region   6816..6830
                     /label=repeat D
                     /note="repeat D"
     misc_feature    6839..6917
                     /label=AT-rich region
                     /note="AT-rich region"
     protein_bind    complement(6929..6937)
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
                     /note="putative DnaA box"
     repeat_region   7033..7039
                     /label=repeat E
                     /note="repeat E"
     repeat_region   7040..7046
                     /label=repeat E
                     /note="repeat E"
     misc_feature    7146..8550
                     /label=similar to GenBank Accession Number M35007
                     /note="similar to GenBank Accession Number M35007"
     regulatory      7277..7282
                     /regulatory_class="minus_35_signal"
     regulatory      7300..7305
                     /regulatory_class="minus_10_signal"
     CDS             7322..8332
                     /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="AAL78957.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            7322..8332
                     /gene="aadA"
                     /label=aadA
     CDS             7541..8332
                     /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       8335..8392
                     /note="palindromic sequence; putative termination signal"
     misc_feature    8551..9152
                     /note="left border region from octopine-type Ti plasmid of 
                     Agrobacterium tumefaciens; similar to GenBank Accession 
                     Number X00493"
     misc_difference 8569..8570
                     /replace="ggtacc"
                     /note="compared to GenBank Accession Number X00493; removed
                     to destroy a KpnI restriction site"
     misc_feature    8852..8876
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    8853..8876
                     /label=left flanking T-DNA border
                     /note="left flanking T-DNA border"