LOCUS       sequence-167727-        5870 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 5870)
  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 5870)
  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..5870
                     /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             1054..1065
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2053..2070)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2223..2309
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2401..2428
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2448..2539
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2540..2608,2609..3400)
                     /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(2758..2777)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3571..4159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4060..4079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4313..4330
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4345..4485
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4411..4430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4571..4593
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4671..4748
                     /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             4749..5831
                     /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(4768..4787)
                     /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 ctgcctgaga attcggatct aggaggatta
      781 tgagatgacc gtttacacag catccgttac cgcacccgtc aacatcgcaa cccttaagta
      841 ttgggggaaa agggacacga agttgaatct gcccaccaat tcgtccatat cagtgacttt
      901 atcgcaagat gacctcagaa cgttgacctc tgcggctact gcacctgagt ttgaacgcga
      961 cactttgtgg ttaaatggag aaccacacag catcgacaat gaaagaactc aaaattgtct
     1021 gcgcgaccta cgccaattaa gaaaggaaat ggaatcgaag gacgcctcat tgcccacatt
     1081 atctcaatgg aaactccaca ttgtctccga aaataacttt cctacagcag ctggtttagc
     1141 ttcctccgct gctggctttg ctgcattggt ctctgcaatt gctaagttat accaattacc
     1201 acagtcaact tcagaaatat ctagaatagc aagaaagggg tctggttcag cttgtagatc
     1261 gttgtttggc ggatacgtgg cctgggaaat gggaaaagct gaagatggtc atgattccat
     1321 ggcagtacaa atcgcagaca gctctgactg gcctcagatg aaagcttgtg tcctagttgt
     1381 cagcgatatt aaaaaggatg tgagttccac tcagggtatg caattgaccg tggcaacctc
     1441 cgaactattt aaagaaagaa ttgaacatgt cgtaccaaag agatttgaag tcatgcgtaa
     1501 agccattgtt gaaaaagatt tcgccacctt tgcaaaggaa acaatgatgg attccaactc
     1561 tttccatgcc acatgtttgg actctttccc tccaatattc tacatgaatg acacttccaa
     1621 gcgtatcatc agttggtgcc acaccattaa tcagttttac ggagaaacaa tcgttgcata
     1681 cacgtttgat gcaggtccaa atgctgtgtt gtactactta gctgaaaatg agtcgaaact
     1741 ctttgcattt atctataaat tgtttggctc tgttcctgga tgggacaaga aatttactac
     1801 tgagcagctt gaggctttca accatcaatt tgaatcatct aactttactg cacgtgaatt
     1861 ggatcttgag ttgcaaaagg atgttgccag agtgatttta actcaagtcg gttcaggccc
     1921 acaagaaaca aacgaatctt tgattgacgc aaagactggt ctaccaaagg aataacccgg
     1981 gaggggatcc tctagagtcg acctgcaggc atgcaagctt ggctgttttg gcggatgaga
     2041 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     2101 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     2161 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     2221 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2281 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     2341 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     2401 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttt gtttattttt
     2461 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2521 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2581 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2641 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2701 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2761 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     2821 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2881 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2941 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3001 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3061 cgagcgtgac accacgatgc ctacagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3121 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3181 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3241 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3301 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3361 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3421 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3481 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3541 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3601 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3661 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3721 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3781 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3841 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3901 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3961 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4021 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4081 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4141 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4201 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4261 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4321 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4381 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4441 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     4501 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4561 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4621 aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg catttacgtt gacaccatcg
     4681 aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     4741 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     4801 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     4861 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     4921 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     4981 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     5041 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     5101 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     5161 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     5221 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     5281 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     5341 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     5401 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     5461 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     5521 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     5581 acagctcatg ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc
     5641 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     5701 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     5761 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5821 gcgggcagtg agcgcaacgc aattaatgtg agttagcgcg aattgatctg
//
