Basic Vector Information
- Vector Name:
- PVRL2
- Antibiotic Resistance:
- Gentamicin
- Length:
- 8722 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.
- Promoter:
- araBAD
PVRL2 vector Vector Map
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
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"
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