pKP77 vector (V005197)

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

pKP7714508 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000FLAGFLAGFLAGcloning siteubiquitinHADHFRFLAGFLAGFLAGUASUbiquitin carboxyl-terminal hydrolase 1tet promoterSm resistance protein BRSF1010 oriVmobilization protein CRSF1010 oriTRSF1010 RepBErepressor protein FRSF1010 RepCNeoR/KanR

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  JOURNAL   Unpublished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            "Unpublished"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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