Basic Vector Information
- Vector Name:
- pKP77
- Antibiotic Resistance:
- Kanamycin
- Length:
- 14508 bp
- Type:
- URT cloning vector
- Replication origin:
- RSF1010 oriV
- Source/Author:
- Piatkov K, Graciet E, Varshavsky A.
- Promoter:
- tet
pKP77 vector Map
pKP77 vector Sequence
LOCUS V005197 14508 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V005197
VERSION V005197
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 14508)
AUTHORS Piatkov K, Graciet E, Varshavsky A.
TITLE Ubiquitin Reference Technique and Its Use in Ubiquitin-Lacking
Prokaryotes
JOURNAL PLoS ONE 8 (6), E67952 (2013)
PUBMED 23825692
REFERENCE 2 (bases 1 to 14508)
AUTHORS Piatkov K, Graciet E, Varshavsky A.
TITLE Broad host range UPR vectors for Gram-negative Bacteria
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 14508)
AUTHORS Piatkov K, Graciet E, Varshavsky A.
TITLE Direct Submission
JOURNAL Submitted (14-JUN-2012) Biology, California Institute of Technology,
1200 E. California Blvd., 147-75, Pasadena, CA 91125, USA
REFERENCE 4 (bases 1 to 14508)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 14508)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2013"; volume: "8"; issue: "6"; pages: "E67952"
SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(14-JUN-2012) Biology, California Institute of Technology, 1200 E.
California Blvd., 147-75, Pasadena, CA 91125, USA"
SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14508
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(10..33)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS complement(46..69)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS complement(82..105)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
misc_feature complement(115..129)
/label="cloning site"
/note="cloning site"
CDS complement(130..357)
/label="ubiquitin"
/note="Saccharomyces cerevisiae ubiquitin"
CDS complement(385..411)
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS complement(418..978)
/label="DHFR"
/note="mouse dihydrofolate reductase"
CDS complement(991..1014)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS complement(1027..1050)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS complement(1063..1086)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
protein_bind 1571..1688
/label="UAS"
/note="upstream activating sequence mediating
Gal4-dependent induction"
CDS 2387..4813
/gene="UBP1"
/label="Ubiquitin carboxyl-terminal hydrolase 1"
/note="Ubiquitin carboxyl-terminal hydrolase 1 from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P25037"
promoter 5096..5124
/label="tet promoter"
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 5336..6139
/codon_start=1
/product="Sm resistance protein A"
/label="Sm resistance protein A"
/protein_id="AFQ55265.1"
/translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
CDS 6139..6975
/codon_start=1
/product="Sm resistance protein B"
/label="Sm resistance protein B"
/protein_id="AFQ55266.1"
/translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
rep_origin complement(7619..8013)
/direction=LEFT
/label="RSF1010 oriV"
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
CDS complement(8040..8324)
/codon_start=1
/product="mobilization protein C"
/label="mobilization protein C"
/protein_id="AFQ55267.1"
/translation="MVKGSNKAADRLAKLEEQRARINAEIQRVRAREQQQERKNETRRK
VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
oriT 8355..8442
/label="RSF1010 oriT"
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 9271..9684
/codon_start=1
/product="mobilization protein B"
/label="mobilization protein B"
/protein_id="AFQ55269.1"
/translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTQQAS
EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
CDS 9681..10649
/label="RSF1010 RepB"
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 10713..10925
/codon_start=1
/product="unknown"
/label="unknown"
/note="E"
/protein_id="AFQ55271.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
CDS 10927..11133
/codon_start=1
/product="repressor protein F"
/label="repressor protein F"
/protein_id="AFQ55272.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS 11163..11999
/label="RSF1010 RepA"
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 11989..12837
/label="RSF1010 RepC"
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(13365..14156)
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
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