LOCUS       40924_27109        7308 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKU1021gps DNA, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7308)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2015"; volume: "112"; issue: "3"; pages: 
            "857-862"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-AUG-2014) Contact:Haruo Ikeda Kitasato University, Kitasato 
            Institute for Life Sciences"; volume: " 1-15-1 Kitasato, Sagamihara,
            Kanagawa 252-0373, Japan URL :http"; pages: 
            "//avermitilis.ls.kitasato-u.ac.j"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             344..1135
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            complement(1476..1585)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1639..1650)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     protein_bind    1721..1820
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     CDS             1830..3644
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
     rep_origin      3867..4455
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /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
                     /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)
                     /gene="rpsJp"
                     /note="promoter of gene encoding ribosomal protein S10;
                     from Streptomyces avermitilis MA-4680"
                     /regulatory_class="promoter"
     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..7269
                     /label=GmR
                     /note="gentamycin acetyltransferase"
ORIGIN
        1 ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc
       61 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
      121 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
      181 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
      241 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
      301 gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
      361 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      421 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
      481 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctccaagacg aggcagcgcg
      541 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
      601 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
      661 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
      721 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
      781 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
      841 gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc gacggcgagg atctcgtcgt
      901 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
      961 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     1021 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     1081 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     1141 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     1201 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccctc
     1261 gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga cggccagcag
     1321 gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc
     1381 tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg tttcatcagc
     1441 catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt
     1501 cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg
     1561 tgggcctact tcacctatcc tgccccgctg acgccgttct cgagcgtgcc ggagcaatcg
     1621 ccctgggtgg gttacacgac gcccctctat ggcccgtact gacggacaca ccgaagcccc
     1681 ggcggcaacc ctcagcggat gccccggggc ttcacgtttt cccaggtcag aagcggtttt
     1741 cgggagtagt gccccaactg gggtaacctt tgagttctct cagttggggg cgtagggtcg
     1801 ccgacatgac acaaggggtt gtgaccgggg tggacacgta cgcgggtgct tacgaccgtc
     1861 agtcgcgcga gcgcgagaac tcgagcgcag caagcccagc gacacagcgt agcgccaacg
     1921 aagacaaggc ggccgacctt cagcgcgaag tcgagcgcga cgggggccgg ttcaggttcg
     1981 tcgggcattt cagcgaagcg ccgggcacgt cggcgttcgg gacggcggag cgcccggagt
     2041 tcgaacgcat cctgaacgaa tgccgcgccg ggcggctcaa catgatcatt gtctatgacg
     2101 tgtcgcgctt ctcgcgcctg aaggtcatgg acgcgattcc gattgtctcg gaattgctcg
     2161 ccctgggcgt gacgattgtt tccactcagg aaggcgtctt ccggcaggga aacgtcatgg
     2221 acctgattca cctgattatg cggctcgacg cgtcgcacaa agaatcttcg ctgaagtcgg
     2281 cgaagattct cgacacgaag aaccttcagc gcgaattggg cgggtacgtc ggcgggaagg
     2341 cgccttacgg cttcgagctt gtttcggaga cgaaggagat cacgcgcaac ggccgaatgg
     2401 tcaatgtcgt catcaacaag ctcgcgcact cgaccactcc ccttaccgga cccttcgagt
     2461 tcgagcccga cgtaatccgg tggtggtggc gtgagatcaa gacgcacaaa caccttccct
     2521 tcaagccggg cagtcaagcc gccattcacc cgggcagcat cacggggctt tgtaagcgca
     2581 tggacgctga cgccgtgccg acccggggcg agacgattgg gaagaagacc gcttcaagcg
     2641 cctgggaccc ggcaaccgtt atgcgaatcc ttcgggaccc gcgtattgcg ggcttcgccg
     2701 ctgaggtgat ctacaagaag aagccggacg gcacgccgac cacgaagatt gagggttacc
     2761 gcattcagcg cgacccgatc acgctccggc cggtcgagct tgattgcgga ccgatcatcg
     2821 agcccgctga gtggtatgag cttcaggcgt ggttggacgg cagggggcgc ggcaaggggc
     2881 tttcccgggg gcaagccatt ctgtccgcca tggacaagct gtactgcgag tgtggcgccg
     2941 tcatgacttc gaagcgcggg gaagaatcga tcaaggactc ttaccgctgc cgtcgccgga
     3001 aggtggtcga cccgtccgca cctgggcagc acgaaggcac gtgcaacgtc agcatggcgg
     3061 cactcgacaa gttcgttgcg gaacgcatct tcaacaagat caggcacgcc gaaggcgacg
     3121 aagagacgtt ggcgcttctg tgggaagccg cccgacgctt cggcaagctc actgaggcgc
     3181 ctgagaagag cggcgaacgg gcgaaccttg ttgcggagcg cgccgacgcc ctgaacgccc
     3241 ttgaagagct gtacgaagac cgcgcggcag gcgcgtacga cggacccgtt ggcaggaagc
     3301 acttccggaa gcaacaggca gcgctgacgc tccggcagca aggggcggaa gagcggcttg
     3361 ccgaacttga agccgccgaa gccccgaagc tcccccttga ccaatggttc cccgaagacg
     3421 ccgacgctga cccgaccggc cctaagtcgt ggtgggggcg cgcgtcagta gacgacaagc
     3481 gcgtgttcgt cgggctcttc gtagacaaga tcgttgtcac gaagtcgact acgggcaggg
     3541 ggcagggaac gcccatcgag aagcgcgctt cgatcacgtg ggcgaagccg ccgaccgacg
     3601 acgacgaaga cgacgcccag gacggcacgg aagacgtagc ggcgtagcga gacacccggg
     3661 aagcctgtta ggcgctgaga cgggcgcaca gcgggcttcc tggggcaatg catcctccag
     3721 cgcggggatc tcatgctgga gttcttcgcc caccccaaaa ggatctaggt gaagatcctt
     3781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     4021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4201 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     4261 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4321 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4381 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4441 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4501 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4561 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4621 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4681 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4741 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     4801 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     4861 gattacgcca agcttgcatg cctgcaggtc gactctagac tacgcatcgc ggtaggcgac
     4921 tctgttggcc atctccatca gttcctcccg ccacacgggg gcgatgggcg cgtcggtgat
     4981 ggccttgcag ccccgctcga cgaggccggc gatgtgctgc tcgacgtcgt cggggacctt
     5041 ggtgtccagg aggtggcggc gcacgtcgtc ggcgctgtgt cccggctcgg tgatcagcgc
     5101 gcggatggat tcgtcgcgct gcatcgccca gcccagcagg agcgtcatct tgtgctgggt
     5161 gaagtcgagt ccggcgggct tgcccgtggc cgcggagtcc ccgaaggcgt ccagcaggtc
     5221 gtcccggagc tggaaggcct cgccgaccgc ctcgccgtac tccgcgaagg cggcgtcgag
     5281 atcggtgcgc cctgccacgt tcgcgccgat gaccaggggc cggtggatgg tgtagcgacc
     5341 ggacttgatg acggcgatca gccgggacag ccgggggtcg accgagaact cggcggccgc
     5401 cacgacgtcc atgtactggc cgatcagtac ctcggagcgc agcttgttcc attccgcgat
     5461 cgccgtcgcc ggcgcctcgc acatcaactc gtcggcgtag gtcagagcga ggccgccgac
     5521 gaggaggccg acgccctcgc cgaaccggcg cgactcaccc tgccagccca ggtcctggtg
     5581 cagcgcggtg tgcttcgcat ggacggtcgg cagcccgcgc cgctggctgg agtcgtccag
     5641 gatgtcgtcg tgcacgagaa gggaggcgtg cagcatctca agggccgcgc cggccgccac
     5701 gaccccggag tcgtcggggt cacccccggc cgcgaggtga ccggcgatac agaaggcggg
     5761 ccggatgcgc ttgccgcccg aggccatcag ctcggccagc gcgtcgatgg gcacgaccgc
     5821 ccgctcgtcc acggccgccc atgtgtcgtg ccctgccgag agcagggact gtatccgggc
     5881 ctccacgcgc cgcagcaggt cggcgctggt gcggcgcacg aggtcgaccg tctcctgact
     5941 ggcgtcgttg gtcacgtccg gagtcatatg tactcagtag tccttcgtct cgataaaacg
     6001 ctctggagcc cgggggttcg cgtcctgacc tccgacccac gcggtcgggt gtgtcgcgct
     6061 cttgctgaca gagatatccg atatggattt ccctaccgag ggacacggcc cgaaaaaaac
     6121 cgcaggaccg ggggcacaac acccaccggg cgcctggtcg gcaccccgct gaaacttccc
     6181 ggaagattcc cgtacgtccg ccccagcgct gcctttcgac agatcgggcg acgagtactg
     6241 tgggactcgc ttccagtcct cccggcggga ggcgcacagc atcggcactc gaccgagcaa
     6301 ctccgacagt ctgccatacg gggcacgggc ctggccaatc gggccgagga tcgtaccccg
     6361 agggtgacgc aggtcaaacc ccgggcactc agcaagggtc gcacaccgat ccgcccgggg
     6421 cacacgcgtc cgaccccggt gccaccggcc ggccgggaac gtgagcacat gtgctcatcc
     6481 cggcagcccc cttccgggtg aacagaacat gtccgtggaa ttggacataa gcctgttcgg
     6541 ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     6601 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     6661 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     6721 tgttatggag cagcaacgat gttacgcagc agcaacgatg ttacgcagca gggcagtcgc
     6781 cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag gctcggccct
     6841 gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt cggagacgta
     6901 gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct ccgtagtaag
     6961 acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct cgcggcttac
     7021 gttctgccca ggtttgagca gccgcgtagt gagatctata tctatgatct cgcagtctcc
     7081 ggcgagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa gcatgaggcc
     7141 aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga tcccgcagtg
     7201 gctctctata caaagttggg catacgggaa gaagtgatgc actttgatat cgacccaagt
     7261 accgccacct aacaattcgt tcaagccgag atcgctagct tcacgctg
//
