LOCUS       40924_46113        7276 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PVRL1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7276)
  AUTHORS   Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.
  TITLE     New shuttle vectors for gene cloning and expression in 
            multidrug-resistant Acinetobacter species
  JOURNAL   Antimicrob. Agents Chemother. (2018) In press
REFERENCE   2  (bases 1 to 7276)
  AUTHORS   Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-NOV-2017) Department of Science, Roma Tre University, 
            Viale Marconi 446, Rome, Roma 00146, Italia
REFERENCE   3  (bases 1 to 7276)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7276)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob.
            Agents Chemother. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-NOV-2017) Department of Science, Roma Tre University, Viale 
            Marconi 446, Rome, Roma 00146, Italia"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(530..1060)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1249..1277)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(1295..1399)
                     /label=AmpR promoter
     primer_bind     1638..1654
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1664..1682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1691..1798
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(1811..1829)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1850..1866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1874..1890)
                     /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(1898..1928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1943..1964)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2252..2840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      3164..3193
                     /label=putative MobA/MobL gene predicted promoter
                     /note="putative MobA/MobL gene predicted promoter"
                     /regulatory_class="promoter"
     CDS             3317..3877
                     /codon_start=1
                     /product="putative MobA/MobL protein"
                     /label=putative MobA/MobL protein
                     /note="contains MobA/MobL family domain; may be involved in
                     plasmid mobilization"
                     /protein_id="AUJ88093.1"
                     /translation="MLTRWDNDQSSLSIYHREIKEEQQEQQRKQQQIERETKQKQEDDK
                     KKRQDYERYLSKYGYVKDAEYDHISDHVASDISMFTRLQAQAWASNDMQEYIRCSKQKY
                     EQISDRIAYERDLKKIGQLSRILDKDRQALADILPQLMPYYEQMQQAVNKRKILLENER
                     QPSFKPRYDQEQPKPKKDNDLTF"
     regulatory      3861..3888
                     /label=putative antitoxin-toxin module-predicted promoter
                     /note="putative antitoxin-toxin module-predicted promoter"
                     /regulatory_class="promoter"
     CDS             3922..4215
                     /codon_start=1
                     /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="AUJ88095.1"
                     /translation="MTEATFTFRVDHDLKQEFSSLAKTVDRSGAQLIRDFMRDFVKKQQ
                     EAADYDKWFKQQVQIGLNEANAGKLIPHEDVKAEFAARRAATLAKLAAKNED"
     CDS             4205..4474
                     /codon_start=1
                     /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="AUJ88096.1"
                     /translation="MKIEWTETARQDRRNIYDYLEERNPIAAIEIDDLIEEKTDLLVDN
                     RLMGRTGRQKDTRELVIHPHYVVVYDITDIIRILRVLHTSQEWS"
     regulatory      4238..5574
                     /note="minimal origin of replication for Acinetobacter sp."
                     /regulatory_class="replication_regulatory_region"
     regulatory      5747..5773
                     /label=putative RCR protein gene predicted promoter
                     /note="putative RCR protein gene predicted promoter"
                     /regulatory_class="promoter"
     CDS             5826..6101
                     /codon_start=1
                     /product="putative RCR protein"
                     /label=putative RCR protein
                     /note="contains a domain belonging to RepB proteins; may be
                     involved in rolling circle replication (RCR)"
                     /protein_id="AUJ88097.1"
                     /translation="MIYYMGVKDMKDPNDQKTQDMLKEPPKTNAERQKAYREKRKSLDS
                     QRLEVFIDKGVSDMLADMVGAAGESQKAILTALIEKEYKRLYAVKK"
     regulatory      6150..6170
                     /note="bidirectional Rho-independent transcriptional
                     terminator"
                     /regulatory_class="terminator"
     CDS             complement(6189..6458)
                     /codon_start=1
                     /product="putative MobA/MobL protein"
                     /label=putative MobA/MobL protein
                     /note="contains MobA/MobL family domain; may be involved in
                     plasmid mobilization"
                     /protein_id="AUJ88098.1"
                     /translation="MVKKTAMELGQMLDEEKEKLERQEKARKDLADAVVKGREQKQARS
                     DDAKRKILIGAYLLKKHDNDIKKLVEQNPDFIGYIRENDKHLFS"
     regulatory      complement(6491..6516)
                     /label=putative MobA/MobL gene predicted promoter
                     /note="putative MobA/MobL gene predicted promoter"
                     /regulatory_class="promoter"
     regulatory      6627..6653
                     /label=putative nickase-like gene predicted promoter
                     /note="putative nickase-like gene predicted promoter"
                     /regulatory_class="promoter"
     CDS             6743..7216
                     /codon_start=1
                     /product="putative nickase-like protein"
                     /label=putative nickase-like protein
                     /note="may be involved in rolling circle replication (RCR)"
                     /protein_id="AUJ88099.1"
                     /translation="MKMAIYHCEMQNISRSDGRSIVACAAYRAGEKLYCDTYGKEQDYT
                     KKTGIEYTQIFAPTGASPDMLDRQTLWNRVEQSELKKNGDIKQEARLAKEVEIALPHEL
                     DKTQRQALVTELCQSLVKAYGVAVDVAIHAPHVHGGRRKKPHAHIRHRRQATC"
ORIGIN
        1 cccctcatgt gcatggggga agtgatgaaa agaaacccat cacgcccaca tactaatgac
       61 aacaagaagc cttacaccgc aagggctagg gaagaaagtc agggagctag atcagaactc
      121 aacgctgaaa aaaatcaggg aaaaagtggc ggatctgacg aacgtccacc tagagcgagc
      181 agggcttgag gtgagagttg atcaccgcag tcataaggat cgtggactag acatcgagcc
      241 gacattcaag gagggacttg aagccaccca cgccaagcgc aggggggagg aactggagat
      301 cgtcaagcgt aatgatgaaa tcaaaaagcg caataaaaaa gctgaaaggc ttgatatgtc
      361 tatcattgag acgagcaaca gagtagagca attgacactt gaacagcagc aacagcagca
      421 ggggctgcat acttggtctg acaatcgatg cgaattggcc gcggcgttgt gacaatttac
      481 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
      541 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
      601 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
      661 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
      721 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
      781 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
      841 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
      901 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
      961 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     1021 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     1081 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     1141 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     1201 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     1261 gaacaggctt atgtcaattc ggttgaatac tcatactctt cctttttcaa tattattgaa
     1321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1441 ttattatcat gacattaacc tataaaaata ggcgtatcac gaccatggtt acaatttcca
     1501 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     1561 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     1621 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     1681 gggcgaattg gagctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct
     1741 gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc ccggtaccca
     1801 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     1861 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1921 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1981 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2041 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2101 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2161 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2221 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2281 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2341 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2401 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2461 ggtatctctg ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2521 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2581 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2641 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2701 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2761 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2821 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2881 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2941 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3001 gtgatccccc ctggaggatg ccgtagacca agcccccgca gtctatgcca ccgcacaaca
     3061 ggcactatac gaccaccaac aacaccgcca aacgcttata gatgtgtatt tgattgaagt
     3121 aaaaacggat gagaagcgag cagagcatag attttacaag gctttgcaag acaatatcga
     3181 ccgcttgcag cttgataaag tgatcaaaga gggacgagca gctacagcac tgctaaggga
     3241 gatcggagag ccactgccgc caccagtgaa agtgaaaaca ggttggttta gtgctgatta
     3301 ttacggcttt gaggacatgc tcacacgttg ggataatgat caatctagcc tttccattta
     3361 tcacagagag ataaaagagg agcaacagga acagcaacga aaacaacagc agatagaaag
     3421 agaaactaaa caaaaacagg aagatgacaa aaagaaacgg caggactatg aaagatattt
     3481 atcaaaatac ggctatgtca aagatgctga gtatgaccat atcagcgacc atgttgcttc
     3541 agacatttcc atgtttacac gtcttcaggc gcaggcatgg gcaagtaacg atatgcaaga
     3601 gtacataagg tgttcaaagc aaaaatacga gcagatcagc gacagaatag cctatgagcg
     3661 tgacttgaaa aaaatcgggc aattgagccg tatcctggat aaagacagac aggcacttgc
     3721 tgatatttta ccgcagttga tgccttacta tgagcagatg caacaggctg ttaataaacg
     3781 gaaaatttta cttgagaatg agcgacagcc atcatttaaa ccacgttatg accaagaaca
     3841 gcctaagccg aaaaaagaca atgacctaac tttttaatgt gttacaatag cattcaaatg
     3901 taacacattg gagtgaaaaa catgacagaa gcgactttta cctttcgggt agaccatgac
     3961 ctaaagcaag aattttcaag ccttgctaag actgttgatc ggtcgggggc gcagctaatc
     4021 cgtgacttca tgcgtgactt tgttaagaag caacaggaag ccgcagacta tgacaaatgg
     4081 ttcaagcagc aggtacagat cggcttaaat gaagccaatg caggcaagtt gattccacat
     4141 gaggacgtaa aggcagagtt tgccgctaga cgtgctgcaa cattggcgaa actggcagcg
     4201 aagaatgaag attgagtgga ctgaaacagc tcgccaagat cgtagaaata tctatgatta
     4261 tcttgaagaa cgcaacccta tagcagctat tgaaattgat gatttaattg aagaaaagac
     4321 agatttactt gttgataatc gactgatggg gcgcacaggc agacagaaag atactaggga
     4381 gttagtgata catccgcatt atgtggttgt atatgacatc actgatataa tacggatact
     4441 cagagtgcta cacacatcgc aggagtggtc atgacttact catgtacttt ggattattta
     4501 gtgttataaa atcctgattt ataaattttt tttgttaaaa aagataaaag ccccttgcaa
     4561 ttgcttgggg ctttaccgta atttatgggg tacagatctt cgatactgac atatcggcaa
     4621 tcgaaagcat taaggtttga cgaccgctaa tgatttcacc acaggggctt aatgtacctg
     4681 tcttaaattc taaggtttta actcgctttg tcaagcatag accccaaaaa tttagccaat
     4741 gtctgtaact caatctgtcc atgtgtgggt gatgaggtac agtgacgcta gcacacatcg
     4801 gaaaaacgct attactaggg gaactgaaca gagtagcgga cgcaatgagt agtcatttaa
     4861 ttggcggtta tgagcgtgtt caggcggtgc tatcaatcgt aatcataaca gtggcagctt
     4921 gatacagtga tgtcatccct gatgcgaaag cgaccgaccg acggtacatc gaatgggaat
     4981 actttagggt gatttttaag aatcgctcta gggtgagtat ttcccattca gctctgctcc
     5041 ctccctctgg tactttaatc aaaagcacta ctaaacatat gtttttaaat aaaaaatatt
     5101 gatatagaga taatattagt aagaataatt aaacaattga atatagataa atcattgtta
     5161 aataaagatt aattattaaa atgaatgtat acttatatat aaatcaatga tttaaaatat
     5221 ttgataaaga aaacttttca aaaaaaatat aattgagatt gtgtcatttc ggtcaattct
     5281 taatatgttc cacgcaagtt ttagctatgg tgctaaacag aaatttgctg aaaaagaact
     5341 tttcactgaa ctggttaaaa tgtaagcagc ctgagagccg ccaaaaattt taaaaacaaa
     5401 ccgccttaat catcttcaaa aaatacctct aaaacctcac catttgcgtt ttaagaccca
     5461 tatttcatcc tgcccttatg ttcccatgct gatagctata aagtgtctgt aatcgcttcc
     5521 tatgacgttc taggctgttg ataacttttg gaacaacgca aaatgttaaa atccgatcat
     5581 tttttaacct agttattttc gttacagggt aactttggta gtattatttc aatatttagt
     5641 tgttacagga taacaaaata tatgttacag ggtaattaat ttagcgttac aggataacta
     5701 tagttacagg gtaacgttaa atagttatcc tgtaatttaa aaatagttgc aggatagcat
     5761 aaatttttac aaataaattt tataaaaagt tatcctgtaa ctaaaaacaa tgttatcctg
     5821 taactatgat atactatatg ggagttaaag acatgaaaga cccgaacgat cagaaaacac
     5881 aggacatgct aaaagagccg ccaaaaacca atgccgaacg acaaaaagca tatcgtgaaa
     5941 aacgcaaaag ccttgatagt caacgtttgg aggtgtttat agataagggt gtgtcggata
     6001 tgcttgcgga catggtgggc gcagcagggg agagccaaaa agccattttg accgcattga
     6061 ttgagaaaga gtataagcgg ctgtatgcag tgaaaaaata gcaagctaaa aaagtacatt
     6121 gattcaaaaa gtaatgcaga taaaaaagaa accctcgatt tatgagggtt ttttataact
     6181 actaatgcct atgaaaataa gtgcttatca ttttctctaa tgtacccaat aaaatcgggg
     6241 ttttgttcca caagtttctt gatgtcatta tcatgtttct tcagtaagta agccccgatc
     6301 aggatcttgc gttttgcatc gtctgacctt gcctgttttt gctctcgacc tttcaccaca
     6361 gcgtcagcta gatcctttct tgccttttcc tgacgttcca atttttcctt ttcttcatcg
     6421 agcatttgac ccaattccat agctgttttt ttgaccattt ttaatgcctg tgtctcaacc
     6481 tattttccga agtatgaacc atcatcaatg tgaaaagtgt tctacttcaa aaactcgact
     6541 tacatcaaga tccgagcgca gcgagcaaaa tcaaaaaaca aagtcaaagc cttattgctc
     6601 ttgcttttgc ttttagcctc gcagagttcc cgaagggcgc acttacgcaa aatttttgct
     6661 acgccaaatt ttgcaagtac ggtcagggaa accccgacac cccaaccgcc caaaacttgg
     6721 ggcggctgtt aataaacagg tgatgaaaat ggcaatttac cattgtgaaa tgcagaacat
     6781 ttcgaggtca gatggtcgct caatcgtggc atgtgcagca taccgagcag gcgaaaaatt
     6841 gtactgtgat acgtacggaa aagagcagga ctacaccaaa aaaacaggca ttgaatacac
     6901 ccaaattttt gccccaactg gggcaagtcc tgacatgtta gatcgtcaaa ccctatggaa
     6961 tcgggtagag caatccgaac taaaaaagaa cggtgacatc aaacaggagg caagattagc
     7021 aaaggaagta gagatagcat tgccgcatga actggataag acacagcgtc aggcacttgt
     7081 taccgagttg tgccagtcct tagttaaagc ctatggagtg gcggtggacg tagcgatcca
     7141 tgcccctcat gtgcatgggg gaaggagaaa gaaacctcac gcccacataa gacacaggcg
     7201 tcaggcaact tgttaaccga gttgtgccag tccttagtta aagcctatgg agtggcggtg
     7261 gacgtagcga tccatg
//
