LOCUS       62056_14365        7545 bp DNA     circular SYN 20-JUL-2019
DEFINITION  Shuttle vector pLPV2Z, complete sequence.
ACCESSION   MK681507
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7545)
  AUTHORS   Visca P.
  TITLE     New shuttle vectors for real-time gene expression analysis in 
            multidrug-resistant Acinetobacter species: in vitro and in vivo 
            applications
  JOURNAL   Appl. Environ. Microbiol. (2019) In press
REFERENCE   2  (bases 1 to 7545)
  AUTHORS   Lucidi M, Prencipe E, Visca P.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-2019) Department of Science, Roma Tre University, 
            Viale Marconi 446, Rome, Roma 00146, Italia
REFERENCE   3  (bases 1 to 7545)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol. (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-MAR-2019) Department of Science, Roma Tre University, Viale 
            Marconi 446, Rome, Roma 00146, Italia"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      194..782
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        974..992
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1022..1038
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1072..1088)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1097..1107
                     /label=stop codons in all the three reading frames
                     /note="stop codons in all the three reading frames"
     regulatory      1107..1117
                     /label=optimized RBS
                     /note="optimized RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1124..4195
                     /label=lacZ
                     /note="beta-galactosidase"
     regulatory      4306..4334
                     /label=ble gene predicted promoter
                     /note="ble gene predicted promoter"
                     /regulatory_class="promoter"
     CDS             4379..4750
                     /label=BleoR
                     /note="antibiotic-binding protein"
     promoter        4927..5031
                     /label=AmpR promoter
     promoter        5049..5077
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5266..5796
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     regulatory      5959..5986
                     /label=putative antitoxin-toxin module-predicted promoter
                     /note="putative antitoxin-toxin module-predicted promoter"
                     /regulatory_class="promoter"
     CDS             6020..6313
                     /codon_start=1
                     /transl_table=11
                     /product="putative PaaA2-like antitoxin"
                     /label=putative PaaA2-like antitoxin
                     /note="putative component of an antitoxin-toxin system
                     involved in plasmid stability and maintenance"
                     /protein_id="QDL88245.1"
                     /translation="MTEATFTFRVDHDLKQEFSSLAKTVDRSGAQLIRDFMRDFVKKQQ
                     EAADYDKWFKQQVQIGLNEANAGKLIPHEDVKAEFAARRAATLAKLAAKNED"
     CDS             6303..6572
                     /codon_start=1
                     /transl_table=11
                     /product="putative ParE2-like toxin"
                     /label=putative ParE2-like toxin
                     /note="putative component of an antitoxin-toxin system
                     involved in plasmid stability and maintenance"
                     /protein_id="QDL88246.1"
                     /translation="MKIEWTETARQDRRNIYDYLEERNPIAAIEIDDLIEEKTDLLVDN
                     RLMGRTGRQKDTRELVIHPHYVVVYDITDIIRILRVLHTSQEWS"
     regulatory      6573..7544
                     /note="minimal origin of replication for Acinetobacter
                     spp."
                     /regulatory_class="replication_regulatory_region"
ORIGIN
        1 tgcatgtcta cggcatcctc caggggggat cacaagttta ctcatatata ctttagattg
       61 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      121 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      181 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      241 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      301 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      361 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      421 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      481 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      541 tggagcgaac gacctacacc gaacagagat acctacagcg tgagctatga gaaagcgcca
      601 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      661 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      721 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      781 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      841 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      901 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      961 aagagcgccc agcaattaac cctcactaaa gggaacaaaa gctgggtacc gggccccccc
     1021 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
     1081 gttctagagc ggccgctaag taagtaaaga ggagaaatta agcatgacca tgattacgga
     1141 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     1201 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1261 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     1321 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     1381 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta
     1441 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     1501 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
     1561 cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg gtcggttacg gccaggacag
     1621 tcgtttgccg tctgaatttg acctgagcgc atttttacgc gccggagaaa accgcctcgc
     1681 ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa gatcaggata tgtggcggat
     1741 gagcggcatt ttccgtgacg tctcgttgct gcataaaccg actacacaaa tcagcgattt
     1801 ccatgttgcc actcgcttta atgatgattt cagccgcgct gtactggagg ctgaagttca
     1861 gatgtgcggc gagttgcgtg actacctacg ggtaacagtt tctttatggc agggtgaaac
     1921 gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta
     1981 tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat
     2041 cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc gacggcacgc tgattgaagc
     2101 agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa
     2161 cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag catcatcctc tgcatggtca
     2221 ggtcatggat gagcagacga tggtgcagga tatcctgctg atgaagcaga acaactttaa
     2281 cgccgtgcgc tgttcgcatt atccgaacca tccgctgtgg tacacgctgt gcgaccgcta
     2341 cggcctgtat gtggtggatg aagccaatat tgaaacccac ggcatggtgc caatgaatcg
     2401 tctgaccgat gatccgcgct ggctaccggc gatgagcgaa cgcgtaacgc gaatggtgca
     2461 gcgcgatcgt aatcacccga gtgtgatcat ctggtcgctg gggaatgaat caggccacgg
     2521 cgctaatcac gacgcgctgt atcgctggat caaatctgtc gatccttccc gcccggtgca
     2581 gtatgaaggc ggcggagccg acaccacggc caccgatatt atttgcccga tgtacgcgcg
     2641 cgtggatgaa gaccagccct tcccggctgt gccgaaatgg tccatcaaaa aatggctttc
     2701 gctacctgga gagacgcgcc cgctgatcct ttgcgaatac gcccacgcga tgggtaacag
     2761 tcttggcggt ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg
     2821 cttcgtctgg gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg
     2881 gtcggcttac ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg
     2941 tctggtcttt gccgaccgca cgccgcatcc agcgctgacg gaagcaaaac accagcagca
     3001 gtttttccag ttccgtttat ccgggcaaac catcgaagtg accagcgaat acctgttccg
     3061 tcatagcgat aacgagctcc tgcactggat ggtggcgctg gatggtaagc cgctggcaag
     3121 cggtgaagtg cctctggatg tcgctccaca aggtaaacag ttgattgaac tgcctgaact
     3181 accgcagccg gagagcgccg ggcaactctg gctcacagta cgcgtagtgc aaccgaacgc
     3241 gaccgcatgg tcagaagccg ggcacatcag cgcctggcag cagtggcgtc tggcggaaaa
     3301 cctcagtgtg acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat
     3361 ggatttttgc atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct
     3421 ttcacagatg tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac
     3481 ccgtgcaccg ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc
     3541 ctgggtcgaa cgctggaagg cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg
     3601 cacggcagat acacttgctg atgcggtgct gattacgacc gctcacgcgt ggcagcatca
     3661 ggggaaaacc ttatttatca gccggaaaac ctaccggatt gatggtagtg gtcaaatggc
     3721 gattaccgtt gatgttgaag tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa
     3781 ctgccagctg gcgcaggtag cagagcgggt aaactggctc ggattagggc cgcaagaaaa
     3841 ctatcccgac cgccttactg ccgcctgttt tgaccgctgg gatctgccat tgtcagacat
     3901 gtataccccg tacgtcttcc cgagcgaaaa cggtctgcgc tgcgggacgc gcgaattgaa
     3961 ttatggccca caccagtggc gcggcgactt ccagttcaac atcagccgct acagtcaaca
     4021 gcaactgatg gaaaccagcc atcgccatct gctgcacgcg gaagaaggca catggctgaa
     4081 tatcgacggt ttccatatgg ggattggtgg cgacgactcc tggagcccgt cagtatcggc
     4141 ggaattccag ctgagcgccg gtcgctacca ttaccagttg gtctggtgtc aaaaataact
     4201 ccatggttct ccttacgcat ctgtgcggta tttcacaccg catacaggtg gcacttttcg
     4261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4321 gctcatgaga caataaccct gataaatgct tcaataatag cacgtgagga gggccaccat
     4381 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
     4441 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
     4501 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
     4561 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
     4621 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
     4681 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
     4741 ggagcaggac tgacacatgc ataccaagta tgcagcccct gctgctgttg ctgctgttca
     4801 agtgtcaatt gctctactct gttgctcgtc tcaatgacca tggtcgtgat acgcctattt
     4861 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4921 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4981 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5041 caaccgaatt gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct
     5101 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     5161 tcatggcttg ttatgactgt ttttttgtac agtctatgcc tcgggcatcc aagcagcaag
     5221 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagc
     5281 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc
     5341 atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat
     5401 cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat
     5461 tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa
     5521 gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag
     5581 atctatatct atgatctcgc agtctccggc gagcaccgga ggcagggcat tgccaccgcg
     5641 ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa
     5701 gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa
     5761 gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca agccgagatc
     5821 ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac gccgcggcca attcgcatcg
     5881 attgtcagac caagtatgca gcccctgctg ctgttgctgc tgatgcatac caagaacagc
     5941 ctaagccgaa aaaagacaat gacctaactt tttaatgtgt tacaatagca ttcaaatgta
     6001 acacattgga gtgaaaaaca tgacagaagc gacttttacc tttcgggtag accatgacct
     6061 aaagcaagaa ttttcaagcc ttgctaagac tgttgatcgg tcgggggcgc agctaatccg
     6121 tgacttcatg cgtgactttg ttaagaagca acaggaagcc gcagactatg acaaatggtt
     6181 caagcagcag gtacagatcg gcttaaatga agccaatgca ggcaagttga ttccacatga
     6241 ggacgtaaag gcagagtttg ccgctagacg tgctgcaaca ttggcgaaac tggcagcgaa
     6301 gaatgaagat tgagtggact gaaacagctc gccaagatcg tagaaatatc tatgattatc
     6361 ttgaagaacg caaccctata gcagctattg aaattgatga tttaattgaa gaaaagacag
     6421 atttacttgt tgataatcga ctgatggggc gcacaggcag acagaaagat actagggagt
     6481 tagtgataca tccgcattat gtggttgtat atgacatcac tgatataata cggatactca
     6541 gagtgctaca cacatcgcag gagtggtcat gacttactca tgtactttgg attatttagt
     6601 gttataaaat cctgatttat aaattttttt tgttaaaaaa gataaaagcc ccttgcaatt
     6661 gcttggggct ttaccgtaat ttatggggta cagatcttcg atactgacat atcggcaatc
     6721 gaaagcatta aggtttgacg accgctaatg atttcaccac aggggcttaa tgtacctgtc
     6781 ttaaattcta aggttttaac tcgctttgtc aagcatagac cccaaaaatt tagccaatgt
     6841 ctgtaactca atctgtccat gtgtgggtga tgaggtacag tgacgctagc acacatcgga
     6901 aaaacgctat tactagggga actgaacaga gtagcggacg caatgagtag tcatttaatt
     6961 ggcggttatg agcgtgttca ggcggtgcta tcaatcgtaa tcataacagt ggcagcttga
     7021 tacagtgatg tcatccctga tgcgaaagcg accgaccgac ggtacatcga atgggaatac
     7081 tttagggtga tttttaagaa tcgctctagg gtgagtattt cccattcagc tctgctccct
     7141 ccctctggta ctttaatcaa aagcactact aaacatatgt ttttaaataa aaaatattga
     7201 tatagagata atattagtaa gaataattaa acaattgaat atagataaat cattgttaaa
     7261 taaagattaa ttattaaaat gaatgtatac ttatatataa atcaatgatt taaaatattt
     7321 gataaagaaa acttttcaaa aaaaatataa ttgagattgt gtcatttcgg tcaattctta
     7381 atatgttcca cgcaagtttt agctatggtg ctaaacagaa atttgctgaa aaagaacttt
     7441 tcactgaact ggttaaaatg taagcagcct gagagccgcc aaaaatttta aaaacaaacc
     7501 gccttaatca tcttcaaaaa atacctctaa aacctcacca tttga
//
