LOCUS Exported 8487 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCWori-A13AMO-aaCPRct 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8487 /organism="synthetic DNA construct" /mol_type="other DNA" 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 605..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(3177..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 4504..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 4613..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 4708..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 //