LOCUS       Exported                7276 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PVRL1, complete sequence.
ACCESSION   MG462882
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7276
                     /organism="Cloning vector PVRL1"
                     /mol_type="other DNA"
                     /note="Acinetobacter sp. - Escherichia coli shuttle vector"
                     /db_xref="taxon:2067737"
     CDS             complement(527..1060)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin 3-N-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance protein"
                     /protein_id="AUJ88094.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(527..1060)
                     /gene="aacC1"
                     /label=aacC1
     regulatory      complement(1249..1277)
                     /regulatory_class="promoter"
                     /note="class 1 integron promoter"
     promoter        complement(1249..1277)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     regulatory      complement(1295..1399)
                     /regulatory_class="promoter"
                     /note="beta-lactamase promoter"
     promoter        complement(1295..1399)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1489..1854)
                     /codon_start=1
                     /product="beta-galactosidase LacZalpha"
                     /label=beta-galactosidase LacZalpha
                     /note="allows blue/white screening in Escherichia coli 
                     DH5alpha"
                     /protein_id="AUJ88092.1"
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     primer_bind     1638..1654
                     /label=M13 forward, universal sequencing primer
                     /note="M13 forward, universal sequencing primer"
     regulatory      1664..1682
                     /regulatory_class="promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1664..1682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1691..1798
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1715..1731
                     /note="SK primer, pBluescript II universal sequencing 
                     primer"
     primer_bind     complement(1765..1781)
                     /note="KS primer, pBluescript II universal sequencing 
                     primer"
     regulatory      complement(1811..1829)
                     /regulatory_class="promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1811..1829)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1850..1866)
                     /label=M13 reverse, universal sequencing primer
                     /note="M13 reverse, universal sequencing primer"
     regulatory      complement(1874..1890)
                     /regulatory_class="other"
                     /note="operator; lac gene operator; lacI repressor binding 
                     site"
     protein_bind    1874..1890
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      complement(1898..1928)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(1898..1928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      complement(1943..1964)
                     /regulatory_class="promoter"
                     /note="CAP binding site"
     protein_bind    1943..1964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      complement(2252..2840)
                     /regulatory_class="replication_regulatory_region"
                     /note="ColE1-like origin of replication"
     rep_origin      complement(2252..2840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      3164..3193
                     /regulatory_class="promoter"
                     /note="putative MobA/MobL gene predicted 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
                     /regulatory_class="promoter"
                     /note="putative antitoxin-toxin module-predicted 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
                     /regulatory_class="replication_regulatory_region"
                     /note="minimal origin of replication for Acinetobacter sp."
     regulatory      5747..5773
                     /regulatory_class="promoter"
                     /note="putative RCR protein gene predicted 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
                     /regulatory_class="terminator"
                     /note="bidirectional Rho-independent transcriptional 
                     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)
                     /regulatory_class="promoter"
                     /note="putative MobA/MobL gene predicted promoter"
     regulatory      6627..6653
                     /regulatory_class="promoter"
                     /note="putative nickase-like gene predicted 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"