LOCUS       sequence-246927-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Chang MC, Eachus RA, Trieu W, Ro DK, Keasling JD
  TITLE     Engineering Escherichia coli for production of functionalized 
            terpenoids using plant P450s.
  JOURNAL   Nat Chem Biol. 2007 May . 3(5):274-7.
  PUBMED    17438551
REFERENCE   2  (bases 1 to 8487)
  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: "Nat Chem 
            Biol. 2007 May . 3(5):274-7."
COMMENT     Sequence Label: pCWori-A13AMO-aaCPRct
FEATURES             Location/Qualifiers
     source          1..8487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        500..604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(605..673,674..1465)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(823..842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1636..2224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2125..2144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2378..2395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2410..2551
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2477..2496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2637..2659
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2672..3133)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVLPRAFR"
     primer_bind     3082..3104
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3096..3113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3097..3113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3153..3169
                     /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)."
     promoter        complement(join(3177..3183,3184..3201,3202..3207))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(3256..4338)
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4300..4319
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4339..4416)
                     /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."
     promoter        join(4504..4509,4510..4527,4528..4534)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    4542..4558
                     /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)."
     promoter        join(4613..4618,4619..4634,4635..4641)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4649..4665
                     /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)."
     promoter        join(4708..4713,4714..4729,4730..4736)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4744..4760
                     /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     complement(8419..8437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agagctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcat cgtcaggtgg
      481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      841 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1141 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1321 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1741 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2341 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2401 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2461 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     2521 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     2581 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     2641 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agaacgccat
     2701 caaaaataat tcgcgtctgg ccttcctgta gccagctttc atcaacatta aatgtgagcg
     2761 agtaacaacc cgtcggattc tccgtgggaa caaacggcgg attgaccgta atgggatagg
     2821 tcacgttggt gtagatgggc gcatcgtaac cgtgcatctg ccagtttgag gggacgacga
     2881 cagtatcggc ctcaggaaga tcgcactcca gccagctttc cggcaccgct tctggtgccg
     2941 gaaaccaggc aaagcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3001 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     3061 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaatccg
     3121 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacatt
     3181 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca
     3241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3301 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     3361 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     3421 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     3481 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     3541 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     3601 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     3661 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     3721 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     3781 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     3841 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     3901 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     3961 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     4021 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     4081 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     4141 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     4201 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     4261 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     4321 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     4381 cataccgcga aaggttttgc accattcgat ggtgtcctgg cacgacaggt ttcccgactg
     4441 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     4501 ggctttacac tttatgcttc cggctcgtat aatgtgtgga attgtgagcg gataacaatt
     4561 tcacacagga aacaggatcg atccatcgat gagcttactc cccatccccc tgttgacaat
     4621 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     4681 aggatcagct tactccccat ccccctgttg acaattaatc atcggctcgt ataatgtgtg
     4741 gaattgtgag cggataacaa tttcacacag gaaacaggat ccatcgatgc ttaggaggtc
     4801 atatgaccgt acacgacatc atcgcaacgt acttcactaa atggtacgta attgtgccgc
     4861 tggcactgat tgcgtatcgc gtgctggatt atttctacgc gacgcgctcc aaaagcacca
     4921 agaaatctct gccagaaccg tggcgcctgc cgatcattgg tcacatgcac cacctgattg
     4981 gtacgacccc gcatcgcggt gttcgtgacc tggcccgtaa atacggctcc ctgatgcacc
     5041 tgcagctggg tgaagtgccg acgatcgttg tctcttcccc gaaatgggcg aaagagatcc
     5101 tgactaccta cgatatcacc ttcgccaacc gtccggaaac tctgaccggc gaaatcgtac
     5161 tgtaccataa cactgatgtt gtgctggcgc cgtatggtga gtactggcgt cagctgcgta
     5221 aaatttgcac tctggaactg ctgagcgtga aaaaggttaa atctttccag tctctgcgcg
     5281 aagaggaatg ctggaacctg gtacaagaga tcaaagcgtc tggctctggc cgtcctgtga
     5341 acctgtccga aaatgtattc aaactgattg ccactatcct gtcccgcgca gcttttggca
     5401 aaggtatcaa agaccagaag gagctgaccg aaatcgtcaa ggaaattctg cgtcagaccg
     5461 gtggctttga tgtagcagat attttcccgt ccaagaaatt tctgcatcac ctgagcggca
     5521 aacgtgcacg cctgacttct ctgcgtaaaa agatcgataa cctgatcgat aatctggtcg
     5581 ccgaacatac tgttaacacc tctagcaaaa ccaacgagac gctgctggac gtactgctgc
     5641 gtctgaaaga ctctgcggaa ttcccgctga cttccgacaa catcaaggct attatcctgg
     5701 acatgtttgg tgccggtact gacacttcct cctctacgat cgaatgggct atcagcgagc
     5761 tgatcaaatg tccgaaggcg atggagaaag tgcaggcgga gctgcgtaaa gctctgaacg
     5821 gtaaagagaa aatccacgag gaagacatcc aggagctgag ctacctgaac atggttatca
     5881 aagaaaccct gcgcctgcat ccaccgctgc cgctggtact gccgcgtgag tgtcgtcaac
     5941 ctgtcaatct ggcgggctac aacatcccga acaaaactaa gctgatcgtt aacgtattcg
     6001 cgattaaccg tgacccagaa tactggaagg atgctgaagc cttcatcccg gaacgctttg
     6061 agaactcctc cgcgactgtg atgggtgcag agtacgagta cctgccgttc ggtgccggtc
     6121 gccgcatgtg tcctggcgcc gcgctgggcc tggctaatgt tcagctgccg ctggctaaca
     6181 ttctgtatca ttttaactgg aaactgccaa atggtgtatc ttatgaccag atcgatatga
     6241 ccgaatcctc tggtgcgacc atgcagcgta aaaccgaact gctgctggtt ccgtcctttt
     6301 aatctagaag gagatatacc atggcgctgg ataaactgga tctgtatgtg atcattacgc
     6361 tggtagttgc ggttgctgct tatttcgcga agaaccgccg tagcagctct gcggccaaga
     6421 aagcagctga atctcctgtt atcgtcgtac caaaaaaagt caccgaggac gaagtagacg
     6481 atggtcgtaa aaaggttacc gtgttcttcg gcacccaaac tggcactgca gaaggcttcg
     6541 cgaaagcgct ggtcgaagaa gctaaggctc gttatgaaaa agcggttttc aaggtaatcg
     6601 atctggatga ctacgctgcg gaagatgacg aatatgaaga aaaactgaag aaagagtctc
     6661 tggccttctt ctttctggct acgtacggtg atggcgaacc gaccgacaac gcagcgcgct
     6721 tctataaatg gtttactgaa ggtgaagaaa aaggcgagtg gctggataag ctgcagtacg
     6781 ctgtgttcgg cctgggtaac cgtcagtacg aacacttcaa caagatcgct aaagttgtcg
     6841 atgaaaaact ggtcgaacaa ggcgctaaac gcctggtccc ggttggcatg ggtgatgacg
     6901 atcaatgcat cgaagatgat ttcaccgcat ggaaagagct ggtttggccg gaactggatc
     6961 agctgctgcg tgacgaagat gatacctccg tggcaacccc gtacactgcc gctgttggcg
     7021 agtaccgtgt agtttttcac gataaaccgg aaacgtacga ccaggaccag ctgaccaacg
     7081 gtcacgctgt tcacgacgcg cagcaccctt gccgctccaa cgtagcagtg aaaaaggaac
     7141 tgcactcccc tctgagcgat cgttcttgta cgcatctgga gttcgacatt agcaataccg
     7201 gcctgtctta cgaaaccggt gaccatgttg gtgtatacgt cgagaacctg tccgaagtgg
     7261 ttgacgaagc agaaaaactg atcggcctgc cgccacacac ctacttctct gtccacaccg
     7321 acaacgaaga tggcacccct ctgggcggtg cgagcctgcc gccgcctttc ccgccttgta
     7381 ccctgcgtaa agccctggct tcctatgcgg atgtactgtc ttctccaaag aaatccgctc
     7441 tgctggctct ggcggcgcat gcgaccgact ccaccgaagc cgaccgtctg aaattcttcg
     7501 cctccccggc aggtaaagac gaatacgcac agtggatcgt tgcttcccac cgctctctgc
     7561 tggaagttat ggaagcattc ccatccgcta aaccgccact gggcgttttc tttgcgagcg
     7621 ttgccccgcg tctgcagccg cgctactaca gcattagcag ctccccgaaa ttcgctccga
     7681 accgcatcca cgttacttgt gctctggtgt atgagcagac tccatccggc cgtgtgcata
     7741 aaggtgtgtg ctccacctgg atgaaaaacg cggtgccgat gaccgaatcc caggactgtt
     7801 cttgggctcc gatttacgtt cgtaccagca acttccgtct gccgtctgac ccgaaggtcc
     7861 cggtgatcat gatcggtccg ggcaccggtc tggccccgtt tcgtggtttt ctgcaggaac
     7921 gtctggccca gaaagaagcg ggcaccgagc tgggcacggc aatcctgttc ttcggctgtc
     7981 gcaatcgtaa agtagacttc atctacgaag atgagctgaa caacttcgtc gagacgggcg
     8041 ctctgtctga actggtgacc gcgttctctc gtgaaggcgc aacgaaagaa tatgtgcagc
     8101 ataaaatgac ccagaaagca tccgacatct ggaacctgct gtctgagggt gcatatctgt
     8161 acgtttgtgg cgacgcgaaa ggtatggcga aagatgtcca ccgtactctg cacactatcg
     8221 tccaggaaca gggcagcctg gacagctcca aagccgaact gtatgtaaag aacctgcaaa
     8281 tggctggccg ctacctgcgt gatgtctggt aagtcgacct gcagcccaag cttatcgatg
     8341 ataagctgtc aaacatgagc agatctgagc ccgcctaatg agcgggcttt tttttcagat
     8401 ctgcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     8461 taatggtttc ttagacgatg cgtcaaa
//
