LOCUS       Exported                7308 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKU1021gps DNA, complete sequence.
ACCESSION   AB982124
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  AUTHORS   Yamada Y, Kuzuyama T, Komatsu M, Shin-Ya K, Omura S, Cane DE, Ikeda 
            H.
  TITLE     Terpene synthases are widely distributed in bacteria
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 112 (3), 857-862 (2015)
  PUBMED    25535391
REFERENCE   2  (bases 1 to 7308)
  AUTHORS   Ikeda H.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-2014) Contact:Haruo Ikeda Kitasato University, 
            Kitasato Institute for Life Sciences; 1-15-1 Kitasato, Sagamihara, 
            Kanagawa 252-0373, Japan URL :http://avermitilis.ls.kitasato-u.ac.jp
REFERENCE   3  (bases 1 to 7308)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7308
                     /organism="Expression vector pKU1021gps"
                     /mol_type="other DNA"
                     /db_xref="taxon:1535211"
     CDS             344..1138
                     /codon_start=1
                     /gene="aphII"
                     /product="aminoglycoside phosphotransferase"
                     /label=aphII
                     /protein_id="BAP82231.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            344..1138
                     /gene="aphII"
                     /label=aphII
     oriT            1476..1585
                     /note="incP origin of transfer"
     oriT            1479..1585
                     /label=oriT of RK2
                     /note="oriT of RK2"
     CDS             complement(1639..1650)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    1721..1820
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     misc_recomb     1751..1791
                     /gene="attP"
                     /note="attachment site; from bacteriophage phiC31"
     gene            1751..1791
                     /gene="attP"
                     /label=attP
     CDS             1806..3647
                     /codon_start=1
                     /gene="int_phiC31"
                     /product="integrase"
                     /label=int_phiC31
                     /note="from bacteriophage phiC31"
                     /protein_id="BAP82232.1"
                     /translation="MTQGVVTGVDTYAGAYDRQSRERENSSAASPATQRSANEDKAADL
                     QREVERDGGRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSR
                     LKVMDAIPIVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILD
                     TKNLQRELGGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPD
                     VIRWWWREIKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWD
                     PATVMRILRDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEP
                     AEWYELQAWLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRK
                     VVDPSAPGQHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEA
                     PEKSGERANLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEER
                     LAELEAAEAPKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTT
                     GRGQGTPIEKRASITWAKPPTDDDEDDAQDGTEDVAA"
     gene            1806..3647
                     /gene="int_phiC31"
                     /label=int_phiC31
     CDS             1830..3647
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     rep_origin      3867..4455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4047..4455
                     /label=pMB1 replication origin
                     /note="pMB1 replication origin"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4779..4809
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4817..4833
                     /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)."
     primer_bind     4841..4857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4900..5967)
                     /codon_start=1
                     /gene="gps"
                     /product="geranyl diphosphate synthase"
                     /label=gps
                     /note="from Streptomyces sp."
                     /protein_id="BAP82233.1"
                     /translation="MTPDVTNDASQETVDLVRRTSADLLRRVEARIQSLLSAGHDTWAA
                     VDERAVVPIDALAELMASGGKRIRPAFCIAGHLAAGGDPDDSGVVAAGAALEMLHASLL
                     VHDDILDDSSQRRGLPTVHAKHTALHQDLGWQGESRRFGEGVGLLVGGLALTYADELMC
                     EAPATAIAEWNKLRSEVLIGQYMDVVAAAEFSVDPRLSRLIAVIKSGRYTIHRPLVIGA
                     NVAGRTDLDAAFAEYGEAVGEAFQLRDDLLDAFGDSAATGKPAGLDFTQHKMTLLLGWA
                     MQRDESIRALITEPGHSADDVRRHLLDTKVPDDVEQHIAGLVERGCKAITDAPIAPVWR
                     EELMEMANRVAYRDA"
     gene            complement(4900..5967)
                     /gene="gps"
                     /label=gps
     regulatory      complement(5971..6517)
                     /regulatory_class="promoter"
                     /gene="rpsJp"
                     /note="promoter of gene encoding ribosomal protein S10; 
                     from Streptomyces avermitilis MA-4680"
     gene            complement(5971..6517)
                     /gene="rpsJp"
                     /label=rpsJp
     promoter        6522..6550
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6739..7272
                     /codon_start=1
                     /gene="aac(3)I"
                     /product="aminoglycoside acetyltransferase"
                     /label=aac(3)I
                     /protein_id="BAP82234.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            6739..7272
                     /gene="aac(3)I"
                     /label=aac(3)I