Basic Vector Information
- Vector Name:
- PVRL1
- Antibiotic Resistance:
- Gentamicin
- Length:
- 7276 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Lucidi M, Runci F, Rampioni G, Frangipani E, Leoni L, Visca P.
- Promoter:
- Pc
PVRL1 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 ℃.
PVRL1 vector Sequence
LOCUS 40924_46113 7276 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PVRL1, complete sequence.
ACCESSION .
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)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7276)
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
(10-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..7276
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(530..1060)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(1249..1277)
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(1295..1399)
/label=AmpR promoter
primer_bind 1638..1654
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1664..1682
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1691..1798
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(1811..1829)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1850..1866)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1874..1890)
/label=lac operator
/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)."
promoter complement(1898..1928)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1943..1964)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2252..2840)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
regulatory 3164..3193
/label=putative MobA/MobL gene predicted promoter
/note="putative MobA/MobL gene predicted promoter"
/regulatory_class="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
/label=putative antitoxin-toxin module-predicted promoter
/note="putative antitoxin-toxin module-predicted promoter"
/regulatory_class="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
/note="minimal origin of replication for Acinetobacter sp."
/regulatory_class="replication_regulatory_region"
regulatory 5747..5773
/label=putative RCR protein gene predicted promoter
/note="putative RCR protein gene predicted promoter"
/regulatory_class="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
/note="bidirectional Rho-independent transcriptional
terminator"
/regulatory_class="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)
/label=putative MobA/MobL gene predicted promoter
/note="putative MobA/MobL gene predicted promoter"
/regulatory_class="promoter"
regulatory 6627..6653
/label=putative nickase-like gene predicted promoter
/note="putative nickase-like gene predicted promoter"
/regulatory_class="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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