LOCUS       Exported                8722 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PVRL2, complete sequence.
ACCESSION   MG551985
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8722
                     /organism="Cloning vector PVRL2"
                     /mol_type="other DNA"
                     /note="Acinetobacter sp. - Escherichia coli shuttle vector"
                     /db_xref="taxon:2067738"
     primer_bind     2..18
                     /note="KS primer, pBluescript II universal sequencing 
                     primer"
     primer_bind     complement(52..68)
                     /note="SK primer, pBluescript II universal sequencing 
                     primer"
     regulatory      complement(101..119)
                     /regulatory_class="promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(101..119)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(129..145)
                     /label=M13 forward, universal sequencing primer
                     /note="M13 forward, universal sequencing primer"
     regulatory      421..507
                     /regulatory_class="terminator"
                     /note="transcriptional terminator T1 from the E. coli rrnB 
                     gene"
     terminator      421..507
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      599..626
                     /regulatory_class="terminator"
                     /note="transcriptional terminator T2 from the E. coli rrnB 
                     gene"
     terminator      599..626
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      723..827
                     /regulatory_class="promoter"
                     /note="beta-lactamase promoter"
     promoter        723..827
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      845..873
                     /regulatory_class="promoter"
                     /note="class 1 integron promoter"
     promoter        845..873
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1062..1595
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin 3-N-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance protein"
                     /protein_id="AUJ88100.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            1062..1595
                     /gene="aacC1"
                     /label=aacC1
     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)
                     /regulatory_class="promoter"
                     /note="putative nickase-like gene predicted promoter"
     regulatory      2882..2907
                     /regulatory_class="promoter"
                     /note="putative MobA/MobL gene predicted 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
                     /regulatory_class="terminator"
                     /note="bidirectional Rho-independent transcriptional 
                     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)
                     /regulatory_class="promoter"
                     /note="putative RCR protein gene predicted promoter"
     regulatory      3824..5160
                     /regulatory_class="replication_regulatory_region"
                     /note="minimal origin of replication for Acinetobacter sp."
     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)
                     /regulatory_class="promoter"
                     /note="putative antitoxin-toxin module predicted 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)
                     /regulatory_class="promoter"
                     /note="putative MobA/MobL gene predicted promoter"
     regulatory      6558..7146
                     /regulatory_class="replication_regulatory_region"
                     /note="ColE1-like origin of replication"
     rep_origin      6558..7146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7503..8381)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose dependent transcriptional regulator"
                     /label=araC
                     /protein_id="AUJ88107.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(7503..8381)
                     /gene="araC"
                     /label=araC
     regulatory      8408..8692
                     /regulatory_class="promoter"
                     /note="PBAD promoter"
     promoter        8408..8692
                     /gene="araBAD"
                     /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
                     /regulatory_class="ribosome_binding_site"
                     /note="modified ribosome binding site"