LOCUS       40924_35843        8732 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Agrobacterium binary vector for plant transformation, with 
            spectinomycin- and gentamycin-resistance genes. The MCS is reversed 
            in pPZP221.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8732)
  AUTHORS   Hajdukiewicz P, Svab Z, Maliga P.
  TITLE     The small, versatile pPZP family of Agrobacterium binary vectors for
            plant transformation.
  JOURNAL   Plant Mol. Biol. 1994;25:989-94.
  PUBMED    7919218
REFERENCE   2  (bases 1 to 8732)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8732)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol. 
            Biol."; date: "1994"; volume: "25"; pages: "989-94"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     The GenBank sequence was corrected by inserting a G at position 
            2249.
FEATURES             Location/Qualifiers
     source          1..8732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1187..1813
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2245..3315
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3384..3578
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3922..4062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4248..4836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5083..5871)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6399..6423
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(6521..6695)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6733..7263)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MVRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7323..8001)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     primer_bind     8230..8246
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(8250..8306)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8316..8332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8340..8356)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(8364..8394)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8409..8430)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    8595..8619
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag
       61 ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg
      121 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac
      181 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc
      241 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc
      301 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc
      361 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg
      421 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc
      481 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg
      541 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc
      601 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg
      661 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg
      721 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa
      781 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc
      841 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg
      901 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg
      961 ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg
     1021 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
     1081 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
     1141 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
     1201 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
     1261 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
     1321 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     1381 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     1441 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     1501 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     1561 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     1621 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     1681 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     1741 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     1801 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     1861 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     1921 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     1981 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     2041 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     2101 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     2161 aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg
     2221 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
     2281 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     2341 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     2401 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
     2461 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     2521 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     2581 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     2641 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     2701 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     2761 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     2821 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     2881 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     2941 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     3001 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     3061 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
     3121 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
     3181 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
     3241 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
     3301 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
     3361 aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
     3421 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
     3481 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
     3541 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
     3601 gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
     3661 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
     3721 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc
     3781 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3841 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3901 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     3961 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     4021 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     4081 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4141 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4201 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4261 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4321 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4381 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4441 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4501 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4561 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4621 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4681 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4741 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4801 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4861 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcatgatata
     4921 tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa taataaaagc agacttgacc
     4981 tgatagtttg gctgtgagca attatgtgct tagtgcatct aatcgcttga gttaacgccg
     5041 gcgaagcggc gtcggcttga acgaatttct agctagacat tatttgccga ctaccttggt
     5101 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     5161 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     5221 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     5281 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     5341 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     5401 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     5461 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     5521 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     5581 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     5641 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     5701 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     5761 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     5821 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccccca tgatgtttaa
     5881 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     5941 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     6001 aaaacatgtc ataacaagaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     6061 cggtcaaggt tctggaccag ttgcgtgacg gcagttacgc tacttgcatt acagcttacg
     6121 aaccgaacga ggcttatgtc cactgggttc gtgcccgaat tgatcacagg cagcaacgct
     6181 ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc
     6241 agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca
     6301 acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt
     6361 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
     6421 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gtactgaatt
     6481 aacgccgaat taattcgggg gatctatagg gactttaggt gatctggatt ttagtactgg
     6541 attttggttt taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat
     6601 actaagggtt tcttatatgc tcaacacgtg agcgaaaccc tataagaacc ctaattccct
     6661 tatctgggaa ctactcacac attattatgg agaaactcga tgtcgatcga ctctagctag
     6721 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
     6781 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
     6841 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
     6901 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
     6961 ctgctcaaac ctgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     7021 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     7081 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     7141 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     7201 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtac
     7261 catggtcgat cgacagatct gcgaatcgag agagatagat ttgtagagag agactggtga
     7321 tttcagcgtg tcctctccaa atgaaatgaa cttccttata tagaggaagg gtcttgcgaa
     7381 ggatagtggg attgtgcgtc atcccttacg tcagtggaga tatcacatca atccacttgc
     7441 tttgaagacg tggttggaac gtcttctttt tccacgatgc tcctcgtggg tgggggtcca
     7501 tctttgggac cactgtcggc agaggcatct tgaacgatag cctttccttt atcgcaatga
     7561 tggcatttgt aggtgccacc ttccttttct actgtccttt tgatgaagtg acagatagct
     7621 gggcaatgga atccgaggag gtttcccgat attacccttt gttgaaaagt ctcaatagcc
     7681 ctttggtctt ctgagactgt atctttgata ttcttggagt agacgagagt gtcgtgctcc
     7741 accatgttat cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc
     7801 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga
     7861 acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact
     7921 gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt
     7981 accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc
     8041 ttggagtaga cgagagtgtc gtgctccacc atgttggcaa gctgctctag cattcgccat
     8101 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8161 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     8221 cacgacgttg taaaacgacg gccagtgcca agcttgcatg cctgcaggtc gactctagag
     8281 gatccccggg taccgagctc gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     8341 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     8401 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     8461 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     8521 ttgcgtattg gctagagcag cttgagcttg gatcagattg tcgtttcccg ccttcagttt
     8581 aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
     8641 tagaataacg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     8701 catgccaacc acagggttcc cctcgggatc aa
//
