LOCUS       40924_46118        8722 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PVRL2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8722)
  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 8722)
  AUTHORS   Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-NOV-2017) Department of Science, Roma Tre University, 
            Viale Marconi 446, Rome, Roma 00146, Italia
REFERENCE   3  (bases 1 to 8722)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8722)
  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 
            (20-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..8722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..18
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(52..68)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(101..119)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(129..145)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      421..507
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      599..626
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        723..827
                     /label=AmpR promoter
     promoter        845..873
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1062..1592
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     CDS             complement(2182..2655)
                     /codon_start=1
                     /product="putative nickase-like protein"
                     /label=putative nickase-like protein
                     /note="may be involved in rolling circle replication (RCR)"
                     /protein_id="AUJ88101.1"
                     /translation="MKMAIYHCEMQNISRSDGRSIVACAAYRAGEKLYCDTYGKEQDYT
                     KKTGIEYTQIFAPTGASPDMLDRQTLWNRVEQSELKKNGDIKQEARLAKEVEIALPHEL
                     DKTQRQALVTELCQSLVKAYGVAVDVAIHAPHVHGGRRKKPHAHIRHRRQATC"
     regulatory      complement(2745..2771)
                     /label=putative nickase-like gene predicted promoter
                     /note="putative nickase-like gene predicted promoter"
                     /regulatory_class="promoter"
     regulatory      2882..2907
                     /label=putative MobA/MobL gene predicted promoter
                     /note="putative MobA/MobL gene predicted promoter"
                     /regulatory_class="promoter"
     CDS             2940..3209
                     /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="AUJ88102.1"
                     /translation="MVKKTAMELGQMLDEEKEKLERQEKARKDLADAVVKGREQKQARS
                     DDAKRKILIGAYLLKKHDNDIKKLVEQNPDFIGYIRENDKHLFS"
     regulatory      3228..3248
                     /note="bidirectional Rho-independent transcriptional
                     terminator"
                     /regulatory_class="terminator"
     CDS             complement(3297..3572)
                     /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="AUJ88103.1"
                     /translation="MIYYMGVKDMKDPNDQKTQDMLKEPPKTNAERQKAYREKRKSLDS
                     QRLEVFIDKGVSDMLADMVGAAGESQKAILTALIEKEYKRLYAVKK"
     regulatory      complement(3625..3651)
                     /label=putative RCR protein gene predicted promoter
                     /note="putative RCR protein gene predicted promoter"
                     /regulatory_class="promoter"
     regulatory      3824..5160
                     /note="minimal origin of replication for Acinetobacter sp."
                     /regulatory_class="replication_regulatory_region"
     CDS             complement(4924..5193)
                     /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="AUJ88104.1"
                     /translation="MKIEWTETARQDRRNIYDYLEERNPIAAIEIDDLIEEKTDLLVDN
                     RLMGRTGRQKDTRELVIHPHYVVVYDITDIIRILRVLHTSQEWS"
     CDS             complement(5183..5476)
                     /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="AUJ88105.1"
                     /translation="MTEATFTFRVDHDLKQEFSSLAKTVDRSGAQLIRDFMRDFVKKQQ
                     EAADYDKWFKQQVQIGLNEANAGKLIPHEDVKAEFAARRAATLAKLAAKNED"
     regulatory      complement(5510..5537)
                     /label=putative antitoxin-toxin module predicted promoter
                     /note="putative antitoxin-toxin module predicted promoter"
                     /regulatory_class="promoter"
     CDS             complement(5521..6081)
                     /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="AUJ88106.1"
                     /translation="MLTRWDNDQSSLSIYHREIKEEQQEQQRKQQQIERETKQKQEDDK
                     KKRQDYERYLSKYGYVKDAEYDHISDHVASDISMFTRLQAQAWASNDMQEYIRCSKQKY
                     EQISDRIAYERDLKKIGQLSRILDKDRQALADILPQLMPYYEQMQQAVNKRKILLENER
                     QPSFKPRYDQEQPKPKKDNDLTF"
     regulatory      complement(6205..6234)
                     /label=putative MobA/MobL gene predicted promoter
                     /note="putative MobA/MobL gene predicted promoter"
                     /regulatory_class="promoter"
     rep_origin      6558..7146
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7506..8381)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        8408..8692
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      8713..8717
                     /label=modified ribosome binding site
                     /note="modified ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
ORIGIN
        1 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
       61 gttctagagc ggccgccacc gcggtggagc tccaattcgc cctatagtga gtcgtattac
      121 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      181 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      241 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaaccatgg
      301 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
      361 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
      421 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
      481 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
      541 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
      601 aaggccatcc tgacggatgg cctttttgcg tttccatggt cgtgatacgc ctatttttat
      661 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      841 cgaattgaca taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg
      901 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
      961 ggcttgttat gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     1021 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg
     1081 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
     1141 ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
     1201 tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc
     1261 tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg
     1321 ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct
     1381 atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca
     1441 tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag
     1501 attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga
     1561 tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct
     1621 tcccggccgc ggagttgttc ggtaaattgt cacaacgccg cggccaattc gcatcgattg
     1681 tcagaccaag tatgcagccc ctgctgctgt tgctgctgtt caagtgtcaa ttgctctact
     1741 ctgttgctcg tctcaatgat agacatatca agcctttcag cttttttatt gcgctttttg
     1801 atttcatcat tacgcttgac gatctccagt tcctcccccc tgcgcttggc gtgggtggct
     1861 tcaagtccct ccttgaatgt cggctcgatg tctagtccac gatccttatg actgcggtga
     1921 tcaactctca cctcaagccc tgctcgctct aggtggacgt tcgtcagatc cgccactttt
     1981 tccctgattt ttttcagcgt tgagttctga tctagctccc tgactttctt ccctagccct
     2041 tgcggtgtaa ggcttcttgt tgtcattagt atgtgggcgt gatgggtttc ttttcatcac
     2101 ttcccccatg cacatgaggg gcatggatcg ctacgtccac cgccactcca taggctttaa
     2161 ctaaggactg gcacaactcg gttaacaagt tgcctgacgc ctgtgtctta tgtgggcgtg
     2221 aggtttcttt ctccttcccc catgcacatg aggggcatgg atcgctacgt ccaccgccac
     2281 tccataggct ttaactaagg actggcacaa ctcggtaaca agtgcctgac gctgtgtctt
     2341 atccagttca tgcggcaatg ctatctctac ttcctttgct aatcttgcct cctgtttgat
     2401 gtcaccgttc ttttttagtt cggattgctc tacccgattc catagggttt gacgatctaa
     2461 catgtcagga cttgccccag ttggggcaaa aatttgggtg tattcaatgc ctgttttttt
     2521 ggtgtagtcc tgctcttttc cgtacgtatc acagtacaat ttttcgcctg ctcggtatgc
     2581 tgcacatgcc acgattgagc gaccatctga cctcgaaatg ttctgcattt cacaatggta
     2641 aattgccatt ttcatcacct gtttattaac agccgcccca agttttgggc ggttggggtg
     2701 tcggggtttc cctgaccgta cttgcaaaat ttggcgtagc aaaaattttg cgtaagtgcg
     2761 cccttcggga actctgcgag gctaaaagca aaagcaagag caataaggct ttgactttgt
     2821 tttttgattt tgctcgctgc gctcggatct tgatgtaagt cgagtttttg aagtagaaca
     2881 cttttcacat tgatgatggt tcatacttcg gaaaataggt tgagacacag gcattaaaaa
     2941 tggtcaaaaa aacagctatg gaattgggtc aaatgctcga tgaagaaaag gaaaaattgg
     3001 aacgtcagga aaaggcaaga aaggatctag ctgacgctgt ggtgaaaggt cgagagcaaa
     3061 aacaggcaag gtcagacgat gcaaaacgca agatcctgat cggggcttac ttactgaaga
     3121 aacatgataa tgacatcaag aaacttgtgg aacaaaaccc cgattttatt gggtacatta
     3181 gagaaaatga taagcactta ttttcatagg cattagtagt tataaaaaac cctcataaat
     3241 cgagggtttc ttttttatct gcattacttt ttgaatcaat gtactttttt agcttgctat
     3301 tttttcactg catacagccg cttatactct ttctcaatca atgcggtcaa aatggctttt
     3361 tggctctccc ctgctgcgcc caccatgtcc gcaagcatat ccgacacacc cttatctata
     3421 aacacctcca aacgttgact atcaaggctt ttgcgttttt cacgatatgc tttttgtcgt
     3481 tcggcattgg tttttggcgg ctcttttagc atgtcctgtg ttttctgatc gttcgggtct
     3541 ttcatgtctt taactcccat atagtatatc atagttacag gataacattg tttttagtta
     3601 caggataact ttttataaaa tttatttgta aaaatttatg ctatcctgca actattttta
     3661 aattacagga taactattta acgttaccct gtaactatag ttatcctgta acgctaaatt
     3721 aattaccctg taacatatat tttgttatcc tgtaacaact aaatattgaa ataatactac
     3781 caaagttacc ctgtaacgaa aataactagg ttaaaaaatg atcggatttt aacattttgc
     3841 gttgttccaa aagttatcaa cagcctagaa cgtcatagga agcgattaca gacactttat
     3901 agctatcagc atgggaacat aagggcagga tgaaatatgg gtcttaaaac gcaaatggtg
     3961 aggttttaga ggtatttttt gaagatgatt aaggcggttt gtttttaaaa tttttggcgg
     4021 ctctcaggct gcttacattt taaccagttc agtgaaaagt tctttttcag caaatttctg
     4081 tttagcacca tagctaaaac ttgcgtggaa catattaaga attgaccgaa atgacacaat
     4141 ctcaattata ttttttttga aaagttttct ttatcaaata ttttaaatca ttgatttata
     4201 tataagtata cattcatttt aataattaat ctttatttaa caatgattta tctatattca
     4261 attgtttaat tattcttact aatattatct ctatatcaat attttttatt taaaaacata
     4321 tgtttagtag tgcttttgat taaagtacca gagggaggga gcagagctga atgggaaata
     4381 ctcaccctag agcgattctt aaaaatcacc ctaaagtatt cccattcgat gtaccgtcgg
     4441 tcggtcgctt tcgcatcagg gatgacatca ctgtatcaag ctgccactgt tatgattacg
     4501 attgatagca ccgcctgaac acgctcataa ccgccaatta aatgactact cattgcgtcc
     4561 gctactctgt tcagttcccc tagtaatagc gtttttccga tgtgtgctag cgtcactgta
     4621 cctcatcacc cacacatgga cagattgagt tacagacatt ggctaaattt ttggggtcta
     4681 tgcttgacaa agcgagttaa aaccttagaa tttaagacag gtacattaag cccctgtggt
     4741 gaaatcatta gcggtcgtca aaccttaatg ctttcgattg ccgatatgtc agtatcgaag
     4801 atctgtaccc cataaattac ggtaaagccc caagcaattg caaggggctt ttatcttttt
     4861 taacaaaaaa aatttataaa tcaggatttt ataacactaa ataatccaaa gtacatgagt
     4921 aagtcatgac cactcctgcg atgtgtgtag cactctgagt atccgtatta tatcagtgat
     4981 gtcatataca accacataat gcggatgtat cactaactcc ctagtatctt tctgtctgcc
     5041 tgtgcgcccc atcagtcgat tatcaacaag taaatctgtc ttttcttcaa ttaaatcatc
     5101 aatttcaata gctgctatag ggttgcgttc ttcaagataa tcatagatat ttctacgatc
     5161 ttggcgagct gtttcagtcc actcaatctt cattcttcgc tgccagtttc gccaatgttg
     5221 cagcacgtct agcggcaaac tctgccttta cgtcctcatg tggaatcaac ttgcctgcat
     5281 tggcttcatt taagccgatc tgtacctgct gcttgaacca tttgtcatag tctgcggctt
     5341 cctgttgctt cttaacaaag tcacgcatga agtcacggat tagctgcgcc cccgaccgat
     5401 caacagtctt agcaaggctt gaaaattctt gctttaggtc atggtctacc cgaaaggtaa
     5461 aagtcgcttc tgtcatgttt ttcactccaa tgtgttacat ttgaatgcta ttgtaacaca
     5521 ttaaaaagtt aggtcattgt cttttttcgg cttaggctgt tcttggtcat aacgtggttt
     5581 aaatgatggc tgtcgctcat tctcaagtaa aattttccgt ttattaacag cctgttgcat
     5641 ctgctcatag taaggcatca actgcggtaa aatatcagca agtgcctgtc tgtctttatc
     5701 caggatacgg ctcaattgcc cgattttttt caagtcacgc tcataggcta ttctgtcgct
     5761 gatctgctcg tatttttgct ttgaacacct tatgtactct tgcatatcgt tacttgccca
     5821 tgcctgcgcc tgaagacgtg taaacatgga aatgtctgaa gcaacatggt cgctgatatg
     5881 gtcatactca gcatctttga catagccgta ttttgataaa tatctttcat agtcctgccg
     5941 tttctttttg tcatcttcct gtttttgttt agtttctctt tctatctgct gttgttttcg
     6001 ttgctgttcc tgttgctcct cttttatctc tctgtgataa atggaaaggc tagattgatc
     6061 attatcccaa cgtgtgagca tgtcctcaaa gccgtaataa tcagcactaa accaacctgt
     6121 tttcactttc actggtggcg gcagtggctc tccgatctcc cttagcagtg ctgtagctgc
     6181 tcgtccctct ttgatcactt tatcaagctg caagcggtcg atattgtctt gcaaagcctt
     6241 gtaaaatcta tgctctgctc gcttctcatc cgtttttact tcaatcaaat acacatctat
     6301 aagcgtttgg cggtgttgtt ggtggtcgta tagtgcctgt tgtgcggtgg catagactgc
     6361 gggggcttgg tctacggcat cctccagggg ggatcacaag tttactcata tatactttag
     6421 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6481 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6541 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6601 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6661 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6721 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6781 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6841 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6901 agcttggagc gaacgaccta caccgaacag agatacctac agcgtgagct atgagaaagc
     6961 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7021 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7081 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7141 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7201 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7261 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7321 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7381 agctggcacg acaggtttcc cgactggaaa gatgcataat gtgcctgtca aatggacgaa
     7441 gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc
     7501 aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg
     7561 ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa
     7621 ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc
     7681 tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa
     7741 agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa
     7801 ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc
     7861 ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga
     7921 tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca
     7981 aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca
     8041 aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac
     8101 tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg
     8161 aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc
     8221 tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa
     8281 aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac
     8341 gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca
     8401 ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac
     8461 tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc
     8521 gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc
     8581 cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga
     8641 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt
     8701 ttttgggcta accttctgaa tc
//
