LOCUS       Exported                5058 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNT562 DNA, complete sequence.
ACCESSION   LC215276
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5058)
  AUTHORS   Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
  TITLE     Characterization of the cryptic plasmid pOfk55 from Legionella 
            pneumophila and construction of a pOfk55-derived shuttle vector
  JOURNAL   Plasmid 90, 30-37 (2017)
  PUBMED    28259635
REFERENCE   2  (bases 1 to 5058)
  AUTHORS   Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2017) Contact:Takashi Nishida Yamaguchi 
            University, The United Graduate School of Veterinary Science; 1677-1
            Yoshida, Yamaguchi, Yamaguchi 753-8515, Japan
REFERENCE   3  (bases 1 to 5058)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5058
                     /organism="Cloning vector pNT562"
                     /mol_type="other DNA"
                     /db_xref="taxon:1953769"
     regulatory      563..572
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1016..1603
                     /codon_start=1
                     /product="putative replication protein"
                     /label=putative replication protein
                     /protein_id="BAX01899.1"
                     /translation="MKKEPIPGKRKNVLAYPENPFWQKTEIKIGSKMVKVSGGKHINIE
                     GESISHSGIHVVKEIDENEFVKIYTKNIKAIFDLKPTAQRVLQYLITELQKTPNADAVY
                     LAWVGAEEYFSENHIKSSRASFHRALSELIQKGFLAESTKPNMFWFNPNLFFNGNRLTF
                     IHEYRKKTIQEIGKEENLVQSDIDKQIQIIFN"
     CDS             complement(1675..2757)
                     /codon_start=1
                     /product="lac repressor"
                     /label=lac repressor
                     /protein_id="BAX01900.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2758..2833)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      3067..3095
                     /regulatory_class="promoter"
                     /note="hybrid between the trp and lac UV5 promoters"
     promoter        3067..3095
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3103..3119
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     misc_feature    3166..3273
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3183..3199
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3233..3249)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3479..4273
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /protein_id="BAX01901.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      4452..5040
                     /direction=RIGHT
                     /label=high-copy-number ColE1 origin of replication
                     /note="high-copy-number ColE1 origin of replication"