PVRL2 vector (V002224)

Basic Vector Information

      • Vector Name:
      • PVRL2
      • Antibiotic Resistance:
      • Gentamicin
      • Length:
      • 8722 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.

PVRL2 vector Vector Map

PVRL28722 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400KS primer, pBluescript II universal sequencing primerSK primer, pBluescript II universal sequencing primerpromoter for bacteriophage T7 RNA polymeraseM13 forward, universal sequencing primertranscriptional terminator T1 from the E. coli rrnB genetranscriptional terminator T2 from the E. coli rrnB genebeta-lactamase promoterclass 1 integron promoteraacC1putative nickase-like proteinputative nickase-like gene predicted promoterputative MobA/MobL gene predicted promoterputative MobA/MobL proteinbidirectional Rho-independent transcriptional terminatorputative RCR proteinputative RCR protein gene predicted promoterminimal origin of replication for Acinetobacter sp.putative PaaA2-like antitoxinputative antitoxin-toxin module predicted promoterputative MobA/MobL gene predicted promoterColE1-like origin of replicationaraCPBAD promotermodified ribosome binding site

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

PVRL2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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"

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