LOCUS Exported 6393 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDN-G1GZmbh SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6393) 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 6393) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6393 /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 317..504 /gene="S. cerevisiae HIS3" /label=HIS3 promoter CDS 505..1167 /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" rep_origin complement(1427..1882) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1564..1585) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1776..1795 /label=F1ori-R /note="F1 origin, reverse primer" terminator 1965..2130 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind 2051..2075 /label=pHybLex-R /note="ADH terminator, reverse primer" protein_bind 2385..2401 /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)." protein_bind 2532..2649 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" protein_bind 2857..2875 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2878..2896 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 2944..2967 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 2997..3710 /codon_start=1 /product="yeast-enhanced green fluorescent protein" /label=yeGFP /note="codons optimized for translation in S. cerevisiae and C. albicans" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" CDS 3735..4109 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" terminator 4142..4331 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(4403..4420) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4574..5162) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4654..4673) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5333..6193) /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 5956..5975 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6194..6298) /gene="bla" /label=AmpR promoter primer_bind 6366..6384 /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 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt 241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta 301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat 361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat 421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag 481 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca 541 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac 601 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg 661 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat 721 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc 781 actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt 841 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg 901 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta 961 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga 1021 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg 1081 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt 1141 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat 1201 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta 1261 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg 1321 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg 1381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt 1441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 1501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 1561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 1621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 1681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 1741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 1801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 1861 aatgcgccgc tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa 1921 gggcgatcgg tgcgggcctc ttcgctatta cgccagctga attggagcga cctcatgcta 1981 tacctgagaa agcaacctga cctacaggaa agagttactc aagaataaga attttcgttt 2041 taaaacctaa gagtcacttt aaaatttgta tacacttatt ttttttataa cttatttaat 2101 aataaaaatc ataaatcata agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc 2161 aacgatttga cccttttcca tcttttcgta aatttctggc aaggtagaca agccgacaac 2221 cttgattgga gacttgacca aacctctggc gaagaattgt taattaagag ctcttaacta 2281 gtgcggccgc cctttagtga gggttgaatt cgaattttca aaaattctta cttttttttt 2341 ggatggacgc aaagaagttt aataatcata ttacatgtgt gaaattgtta tccgctcaca 2401 attccacaca acatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat 2461 cttagcctaa aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct 2521 atattgaagt acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc 2581 ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc 2641 actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat 2701 tggcagtaac ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat 2761 gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt 2821 tttgatctat taacagatat ataaatgcaa aaactgtccc tatcagtgat agagatctcc 2881 ctatcagtga tagagatatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt 2941 gttaatatac ctctatactt taacgtcaag gagaaaaaac cccggatcct attaaaatgt 3001 ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg 3061 ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat 3121 tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca accttagtca 3181 ctactttcgg ttatggtgtt caatgttttg cgagataccc agatcatatg aaacaacatg 3241 actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag 3301 atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata 3361 gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg 3421 aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca 3481 aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt 3541 atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat 3601 ccactcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag 3661 aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa ggtgacggtg 3721 ctggtttaat taacatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg 3781 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg 3841 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc 3901 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg 3961 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg 4021 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg 4081 tgcacttcgt ggccgaggag caggactgac tcgagtaagc ttggtaccgc ggctagctaa 4141 gatccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt 4201 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc 4261 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg 4321 gacgctcgaa gatccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4381 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4441 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4501 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4981 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 5041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5101 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5161 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5221 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5281 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5341 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5401 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 5461 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 5521 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 5581 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 5641 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 5701 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 5761 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 5821 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 5881 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 5941 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6001 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6061 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6121 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6181 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6241 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6301 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 6361 ataaaaatag gcgtatcacg aggccctttc gtc //