LOCUS Exported 5958 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDN-NG1Tt SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5958) AUTHORS Nevozhay D, Adams RM, Murphy KF, Josic K, Balazsi G TITLE Negative autoregulation linearizes the dose-response and suppresses the heterogeneity of gene expression. JOURNAL Proc Natl Acad Sci U S A. 2009 Mar 31;106(13):5123-8. doi: 10.1073/pnas.0809901106. Epub 2009 Mar 11. PUBMED 19279212 REFERENCE 2 (bases 1 to 5958) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5958 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 191..472 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 473..1147 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" rep_origin complement(1246..1701) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1383..1404) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1595..1614 /label=F1ori-R /note="F1 origin, reverse primer" terminator 1779..1944 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind 1865..1889 /label=pHybLex-R /note="ADH terminator, reverse primer" protein_bind 2439..2455 /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 2586..3027 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 2586..2703 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 2998..3021 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 3051..3674 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind complement(3124..3143) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" terminator 3707..3896 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(3968..3985) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4139..4727) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4219..4238) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4898..5758) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5521..5540 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5759..5863) /gene="bla" /label=AmpR promoter primer_bind 5931..5949 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatagatc aacgacatta ctatatatat aatataggaa gcatttaata gaacagcatc 241 gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag atattcttta 301 ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct ttttttgccg 361 attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg 421 actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt 481 tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga 541 ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat gtgtgcccaa 601 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata 661 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga 721 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga 781 gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc 841 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt 901 gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg 961 ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg gactgacgcc 1021 agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg 1081 tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa 1141 gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt gcctatgcgg 1201 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaaacgtta 1261 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg 1321 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg 1381 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa 1441 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg 1501 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt 1561 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg 1621 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta 1681 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg 1741 atcggtgcgg gcctcttcgc tattacgcca ctgaattgga gcgacctcat gctatacctg 1801 agaaagcaac ctgacctaca ggaaagagtt actcaagaat aagaattttc gttttaaaac 1861 ctaagagtca ctttaaaatt tgtatacact tatttttttt ataacttatt taataataaa 1921 aatcataaat cataagaaat tcgcttattt agaagtgtca acaacgtatc taccaacgat 1981 ttgacccttt tccatctttt cgtaaatttc tggcaaggta gacaagccga caaccttgat 2041 tggagacttg accaaacctc tggcgaagaa ttgttaatta agagctctta agacccactt 2101 tcacatttaa gttgtttttc taatccgcat atgatcaatt caaggccgaa taagaaggct 2161 ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg cggcatacta 2221 tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc ttccaatacg 2281 caacctaaag taaaatgccc cacagcgctg agtgcatata atgcattctc actagtgcgg 2341 ccgcccttta gtgagggttg aattcgaatt ttcaaaaatt cttacttttt ttttggatgg 2401 acgcaaagaa gtttaataat catattacat gtgtgaaatt gttatccgct cacaattcca 2461 cacaacatat ctaatcttac ttatatgttg tggaaatgta aagagcccca ttatcttagc 2521 ctaaaaaaac cttctctttg gaactttcag taatacgctt aactgctcat tgctatattg 2581 aagtacggat tagaagccgc cgagcgggtg acagccctcc gaaggaagac tctcctccgt 2641 gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct 2701 ccgaacaata aagattctac aatactagct tttatggtta tgaagaggaa aaattggcag 2761 taacctggcc ccacaaacct tcaaatgaac gaatcaaatt aacaaccata ggatgataat 2821 gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat gatttttgat 2881 ctattaacag atatataaat gcaaaaactg cataaccact ttaactaata ctttcaacat 2941 tttcggtttg tattacttct tattcaaatg taataaaagt atcaacaaaa aattgttaat 3001 atacctctat actttaacgt caaggagaaa aaaccccgga tcctattaaa atgtctagat 3061 tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag 3121 gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc 3181 atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc 3241 atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta 3301 aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac 3361 ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag 3421 gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt 3481 gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg 3541 atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc 3601 cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg 3661 aaagtgggtc ttaactcgag taagcttggt accgcggcta gctaagatcc gctctaaccg 3721 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt 3781 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt 3841 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaagatcc 3901 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 3961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 4501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 4561 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 4621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 4681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4801 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4861 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 4921 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4981 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5041 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5101 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5161 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5221 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5281 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5341 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5401 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5461 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5521 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5581 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5641 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5701 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5761 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5821 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 5881 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 5941 tcacgaggcc ctttcgtc //