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PVRL1 vector (V002225#)

Basic Vector Information

Basic Vector Information
Vector Name PVRL1 Antibiotic Resistance Gentamycin
Length 7276 bp Type Cloning vector
Source Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.

PVRL1 vector Vector Map

aacC1 class 1 integron promoter beta-lactamase promoter beta-galactosidase LacZalpha operator lac promoter CAP binding site ColE1-like origin of replication putative MobA/MobL gene predicted promoter putative MobA/MobL protein putative PaaA2-like antitoxin minimal origin of replication for Acinetobacter sp. putative RCR protein gene predicted promoter putative RCR protein bidirectional Rho-independent transcriptional terminator putative MobA/MobL protein putative MobA/MobL gene predicted promoter putative nickase-like gene predicted promoter putative nickase-like protein

PVRL1 vector Sequence

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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"

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