LOCUS       Exported                7254 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKU1021fps DNA, complete sequence.
ACCESSION   AB982125
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7254)
  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 7254)
  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 7254)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7254
                     /organism="Expression vector pKU1021fps"
                     /mol_type="other DNA"
                     /db_xref="taxon:1535212"
     CDS             353..1147
                     /codon_start=1
                     /gene="aphII"
                     /product="aminoglycoside phosphotransferase"
                     /label=aphII
                     /protein_id="BAP82235.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            353..1147
                     /gene="aphII"
                     /label=aphII
     oriT            1485..1594
                     /note="incP origin of transfer"
     oriT            1488..1594
                     /label=oriT of RK2
                     /note="oriT of RK2"
     CDS             complement(1648..1659)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    1730..1829
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     misc_recomb     1760..1800
                     /gene="attP"
                     /note="attachment site; from bacteriophage phiC31"
     gene            1760..1800
                     /gene="attP"
                     /label=attP
     CDS             1815..3656
                     /codon_start=1
                     /gene="int_phiC31"
                     /product="integrase"
                     /label=int_phiC31
                     /note="from bacteriophage phiC31"
                     /protein_id="BAP82236.1"
                     /translation="MTQGVVTGVDTYAGAYDRQSRERENSSAASPATQRSANEDKAADL
                     QREVERDGGRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSR
                     LKVMDAIPIVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILD
                     TKNLQRELGGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPD
                     VIRWWWREIKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWD
                     PATVMRILRDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEP
                     AEWYELQAWLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRK
                     VVDPSAPGQHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEA
                     PEKSGERANLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEER
                     LAELEAAEAPKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTT
                     GRGQGTPIEKRASITWAKPPTDDDEDDAQDGTEDVAA"
     gene            1815..3656
                     /gene="int_phiC31"
                     /label=int_phiC31
     CDS             1839..3656
                     /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      3876..4464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4056..4464
                     /label=pMB1 replication origin
                     /note="pMB1 replication origin"
     protein_bind    4752..4773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4788..4818
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4826..4842
                     /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     4850..4866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4909..5922)
                     /codon_start=1
                     /gene="ptlB"
                     /product="farnesyl diphosphate synthase"
                     /label=ptlB
                     /note="from Streptomyces avermitilis"
                     /protein_id="BAP82237.1"
                     /translation="MTVTPESLHRSRDAITPALRTALHLLDEETRHQAAYHLGWIEADG
                     TPADGGGGKAVRPALALLSAQAAGAPAETGLPGAVAVELVHNFSLLHDDLMDGDEERRH
                     RRTVWTLWGASGAILTGDALLALAQEVLLDCGLPTGAAAARLLARTTRDLIRGQVQDVA
                     FERRAHVPIEECVDMAAGKTGALLSASAAIGAVLAGAPVTTVDALALYGQQVGLAFQLT
                     DDVLGIWGDPAVTGKSVHSDLRARKKSLPVTYALNQGGTVSVELATWLSAPGEPDEAEL
                     RRAADLVAAAGGRDWATAEAARRMALGEAALRGVDLDAAARDELIALGHFVVDREV"
     gene            complement(4909..5922)
                     /gene="ptlB"
                     /label=ptlB
     regulatory      complement(5926..6472)
                     /regulatory_class="promoter"
                     /gene="rpsJp"
                     /note="promoter of gene encoding ribosomal protein S10; 
                     from Streptomyces avermitilis MA-4680"
     gene            complement(5926..6472)
                     /gene="rpsJp"
                     /label=rpsJp
     promoter        6477..6505
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6694..7227
                     /codon_start=1
                     /gene="aac(3)I"
                     /product="aminoglycoside acetyltransferase"
                     /label=aac(3)I
                     /protein_id="BAP82238.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            6694..7227
                     /gene="aac(3)I"
                     /label=aac(3)I