LOCUS       sequence-163506-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NudB (E. coli) and PMD (S. cerevisiae) are under 
            control of Trc promoter for isopentenol production..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  AUTHORS   George KW, Thompson MG, Kang A, Baidoo E, Wang G, Chan LJ, Adams PD,
            Petzold CJ, Keasling JD, Lee TS
  TITLE     Metabolic engineering for the high-yield production of 
            isoprenoid-based C(5) alcohols in E. coli.
  JOURNAL   Sci Rep. 2015 Jun 8;5:11128. doi: 10.1038/srep11128.
  PUBMED    26052683
REFERENCE   2  (bases 1 to 5855)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2015-06-8"; volume: "5:11128. doi"; pages: " 
            10.1038/srep11128"
COMMENT     Sequence Label: JBEI-6833
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1066..1077
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2038..2055)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2208..2294
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2386..2413
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2433..2524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2525..2593,2594..3385)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2743..2762)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3556..4144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4045..4064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4298..4315
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4330..4470
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4396..4415)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4556..4578
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4656..4733
                     /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."
     CDS             4734..5816
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4753..4772)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg gaattcaaaa gatcttttaa gaaggagata
      301 tacatatgga ggataaagtg tataagcgtc ccgtttcgat cttagtggtc atctacgcac
      361 aagatacgaa acgggtgctg atgttgcagc ggcgtgacga tcccgatttc tggcagtcgg
      421 taaccggcag cgtggaagag ggtgaaaccg cgccgcaagc tgccatgcgc gaagtaaagg
      481 aagaggtcac cattgatgtt gtcgctgaac aactgacctt aattgactgt cagcgcacgg
      541 tagagtttga aattttttca catttacgtc atcgctatgc gccgggcgtg acgcgtaata
      601 cggaatcatg gttctgtctt gcgcttccgc acgagcggca gatcgttttc actgaacatc
      661 tggcttacaa gtggcttgat gcgcctgctg cggcggcgct cactaagtcc tggagcaacc
      721 ggcaggcgat tgaacagttt gtaattaacg ctgcctgagg atccaaactc gagttcggat
      781 ctaggaggat tatgagatga ccgtttacac agcatccgtt accgcacccg tcaacatcgc
      841 aacccttaag tattggggga aaagggacac gaagttgaat ctgcccacca attcgtccat
      901 atcagtgact ttatcgcaag atgacctcag aacgttgacc tctgcggcta ctgcacctga
      961 gtttgaacgc gacactttgt ggttaaatgg agaaccacac agcatcgaca atgaaagaac
     1021 tcaaaattgt ctgcgcgacc tacgccaatt aagaaaggaa atggaatcga aggacgcctc
     1081 attgcccaca ttatctcaat ggaaactcca cattgtctcc gaaaataact ttcctacagc
     1141 agctggttta gcttcctccg ctgctggctt tgctgcattg gtctctgcaa ttgctaagtt
     1201 ataccaatta ccacagtcaa cttcagaaat atctagaata gcaagaaagg ggtctggttc
     1261 agcttgtaga tcgttgtttg gcggatacgt ggcctgggaa atgggaaaag ctgaagatgg
     1321 tcatgattcc atggcagtac aaatcgcaga cagctctgac tggcctcaga tgaaagcttg
     1381 tgtcctagtt gtcagcgata ttaaaaagga tgtgagttcc actcagggta tgcaattgac
     1441 cgtggcaacc tccgaactat ttaaagaaag aattgaacat gtcgtaccaa agagatttga
     1501 agtcatgcgt aaagccattg ttgaaaaaga tttcgccacc tttgcaaagg aaacaatgat
     1561 ggattccaac tctttccatg ccacatgttt ggactctttc cctccaatat tctacatgaa
     1621 tgacacttcc aagcgtatca tcagttggtg ccacaccatt aatcagtttt acggagaaac
     1681 aatcgttgca tacacgtttg atgcaggtcc aaatgctgtg ttgtactact tagctgaaaa
     1741 tgagtcgaaa ctctttgcat ttatctataa attgtttggc tctgttcctg gatgggacaa
     1801 gaaatttact actgagcagc ttgaggcttt caaccatcaa tttgaatcat ctaactttac
     1861 tgcacgtgaa ttggatcttg agttgcaaaa ggatgttgcc agagtgattt taactcaagt
     1921 cggttcaggc ccacaagaaa caaacgaatc tttgattgac gcaaagactg gtctaccaaa
     1981 ggaataactg caggcatgca agcttggctg ttttggcgga tgagagaaga ttttcagcct
     2041 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     2101 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     2161 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     2221 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     2281 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     2341 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     2401 cggatggcct ttttgcgttt ctacaaactc tttttgttta tttttctaaa tacattcaaa
     2461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2521 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2581 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2641 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2701 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2761 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2821 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2881 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2941 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3001 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3061 gatgcctaca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3121 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3181 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3241 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3301 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3361 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3661 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4201 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4261 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4321 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4381 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4441 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4501 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4561 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat
     4621 tcgcgcgcga aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct
     4681 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     4741 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     4801 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     4861 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     4921 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     4981 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5041 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5101 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5161 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5221 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     5281 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     5341 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     5401 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     5461 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     5521 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     5581 tcccgccgtc aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     5641 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     5701 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5761 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5821 aacgcaatta atgtgagtta gcgcgaattg atctg
//
