LOCUS       Exported                8732 bp ds-DNA    circular SYN 11-1-2016
DEFINITION  Agrobacterium binary vector for plant transformation, with 
            spectinomycin- and gentamycin-resistance genes. The MCS is reversed 
            in pPZP221.
ACCESSION   U10463
VERSION     .
KEYWORDS    pPZP222
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     The GenBank sequence was corrected by inserting a G at position 
            2249.
FEATURES             Location/Qualifiers
     source          1..8732
                     /organism="synthetic DNA construct"
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
     CDS             1187..1816
                     /codon_start=1
                     /product="stability protein from Pseudomonas plasmid pVS1"
                     /note="pVS1 StaA"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2245..3318
                     /codon_start=1
                     /product="replication protein from Pseudomonas plasmid 
                     pVS1"
                     /note="pVS1 RepA"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3384..3578
                     /note="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3922..4062
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4248..4836)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5080..5871)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase"
                     /note="SmR"
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6399..6423
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    6521..6695
                     /note="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6730..7263)
                     /codon_start=1
                     /gene="aacC1"
                     /allele="
                     "
                     /product="gentamycin acetyltransferase"
                     /note="GmR"
                     /note="confers resistance to gentamycin"
                     /translation="MVRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7323..8001)
                     /note="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             complement(8087..8320)
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     primer_bind     8230..8246
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8250..8306
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC18 multiple cloning site"
     primer_bind     complement(8316..8332)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8340..8356
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     misc_feature    8595..8619
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"