LOCUS Exported 7356 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDZ513 (pHIS MET25pro PP7-PS-yeGFP) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7356) AUTHORS Hocine S, Raymond P, Zenklusen D, Chao JA, Singer RH TITLE Single-molecule analysis of gene expression using two-color RNA labeling in live yeast. JOURNAL Nat Methods. 2013 Feb;10(2):119-21. doi: 10.1038/nmeth.2305. Epub 2012 Dec 23. PUBMED 23263691 REFERENCE 2 (bases 1 to 7356) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7356 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 62..411 /gene="S. cerevisiae MET17" /gene_synonym="MET15; MET25" /label=MET17 promoter /note="expression is repressed by methionine" CDS 817..1527 /codon_start=1 /product="yeast-enhanced green fluorescent protein" /label=yeGFP /note="codons optimized for translation in S. cerevisiae and C. albicans" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" CDS 1537..2250 /codon_start=1 /product="yeast-enhanced green fluorescent protein" /label=yeGFP /note="codons optimized for translation in S. cerevisiae and C. albicans" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" terminator 2260..2507 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(2276..2294) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(2525..2544) /label=T7 /note="T7 promoter, forward primer" promoter complement(2526..2544) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2554..2571) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2554..2570) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2563..2585) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2715..3170 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2802..2821) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3012..3033 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(3430..4089) /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" promoter complement(4090..4276) /gene="S. cerevisiae HIS3" /label=HIS3 promoter primer_bind complement(4423..4442) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4542..4564 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4602..4620) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 4662..5165 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 5202..5306 /gene="bla" /label=AmpR promoter CDS 5307..6167 /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 complement(5525..5544) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6338..6926 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6827..6846 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7080..7097 /label=L4440 /note="L4440 vector, forward primer" promoter 7250..7280 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7288..7304 /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 7293..7315 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7312..7328 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7312..7328 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7347..11 /label=T3 /note="T3 promoter, forward primer" promoter 7349..11 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ctcactaaag ggaacaaaag ctggagctcc ggatgcaagg gttcgaatcc cttagctctc 61 attatttttt gctttttctc ttgaggtcac atgatcgcaa aatggcaaat ggcacgtgaa 121 gctgtcgata ttggggaact gtggtggttg gcaaatgact aattaagtta gtcaaggcgc 181 catcctcatg aaaactgtgt aacataataa ccgaagtgtc gaaaaggtgg caccttgtcc 241 aattgaacac gctcgatgaa aaaaataaga tatatataag gttaagtaaa gcgtctgtta 301 gaaaggaagt ttttcctttt tcttgctctc ttgtcttttc atctactatt tccttcgtgt 361 aatacagggt cgtcagatac atagatacaa ttctattacc cccatccata ctctagaatg 421 tccaaaacca tcgttctttc ggtcggcgag gctactcgca ctctgactga gatccagtcc 481 accgcagacc gtcagatctt cgaagagaag gtcgggcctc tggtgggtcg gctgcgcctc 541 acggcttcgc tccgtcaaaa cggagccaag accgcgtatc gcgtcaacct aaaactggat 601 caggcggacg tcgttgattc cggacttccg aaagtgcgct acactcaggt atggtcgcac 661 gacgtgacaa tcgttgcgaa tagcaccgag gcctcgcgca aatcgttgta cgatttgacc 721 aagtccctcg tcgcgacctc gcaggtcgaa gatcttgtcg tcaaccttgt gccgctgggc 781 cgtggatccc ccgggctgca ggaattcgat atcgtgtcta aaggtgaaga attattcact 841 ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc 901 tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt tatttgtact 961 actggtaaat tgccagttcc atggccaacc ttagtcacta ctttcggtta tggtgttcaa 1021 tgttttgcga gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca 1081 gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga 1141 gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat 1201 tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa ctctcacaat 1261 gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa aattagacac 1321 aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac tccaattggt 1381 gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc cttatccaaa 1441 gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc tgctggtatt 1501 acccatggta tggatgaatt gtacaaagat atcgtgtcta aaggtgaaga attattcact 1561 ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc 1621 tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt tatttgtact 1681 actggtaaat tgccagttcc atggccaacc ttagtcacta ctttcggtta tggtgttcaa 1741 tgttttgcga gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca 1801 gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga 1861 gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat 1921 tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa ctctcacaat 1981 gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa aattagacac 2041 aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac tccaattggt 2101 gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc cttatccaaa 2161 gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc tgctggtatt 2221 acccatggta tggatgaatt gtacaaataa ctggtcgagt catgtaatta gttatgtcac 2281 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac 2341 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta 2401 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga 2461 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa 2521 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga 2581 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 2641 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 2701 tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2761 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2821 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 2881 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 2941 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 3001 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 3061 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 3121 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg 3181 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagatccgtc gagttcaaga 3241 gaaaaaaaaa gaaaaagcaa aaagaaaaaa ggaaagcgcg cctcgttcag aatgacacgt 3301 atagaatgat gcattacctt gtcatcttca gtatcatact gttcgtatac atacttactg 3361 acattcatag gtatacatat atacacatgt atatatatcg tatgctgcag ctttaaataa 3421 tcggtgtcac tacataagaa cacctttggt ggagggaaca tcgttggtac cattgggcga 3481 ggtggcttct cttatggcaa ccgcaagagc cttgaacgca ctctcactac ggtgatgatc 3541 attcttgcct cgcagacaat caacgtggag ggtaattctg ctagcctctg caaagctttc 3601 aagaaaatgc gggatcatct cgcaagagag atctcctact ttctcccttt gcaaaccaag 3661 ttcgacaact gcgtacggcc tgttcgaaag atctaccacc gctctggaaa gtgcctcatc 3721 caaaggcgca aatcctgatc caaacctttt tactccacgc gccagtaggg cctctttaaa 3781 agcttgaccg agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc 3841 accaatgcac tcaacgatta gcgaccagcc ggaatgcttg gccagagcat gtatcatatg 3901 gtccagaaac cctatacctg tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc 3961 tactgcttct gcctcttttt ctgggaagat cgagtgctct atcgctaggg gaccaccctt 4021 taaagagatc gcaatctgaa tcttggtttc atttgtaata cgctttacta gggctttctg 4081 ctctgtcatc tttgccttcg tttatcttgc ctgctcattt tttagtatat tcttcgaaga 4141 aatcacatta ctttatataa tgtataattc attatgtgat aatgccaatc gctaagaaaa 4201 aaaaagagtc atccgctagg ggaaaaaaaa aaatgaaaat cattaccgag gcataaaaaa 4261 atatagagtg tactagagga ggccaagagt aatagaaaaa gaaaattgcg ggaaaggact 4321 gtgttatgac ttccctgact aatgccgtgt tcaaacgata cctggcagtg actcctagcg 4381 ctcaccaagc tcttaaaacg ggaatttatg gtgcactctc agtacaatct gctctgatgc 4441 cgcatagtta agccagcccc gacacccgcc aacacgcgct gacgcgccct gacgggcttg 4501 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 4561 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt 4621 tttataggtt aatgtcatga taataatggt ttcttaggac ggatcgcttg cctgtaactt 4681 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta 4741 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa 4801 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat 4861 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa 4921 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg 4981 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct 5041 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt 5101 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac 5161 gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 5221 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 5281 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 5341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 5401 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 5461 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 5521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 5581 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 5641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 5701 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 5761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 5821 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 5881 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 5941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 6001 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 6061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 6121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 6181 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 6241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 6301 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 6361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 6421 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 6481 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 6541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 6601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 6661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 6721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 6781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 6841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 6901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 6961 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 7021 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 7081 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc 7141 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 7201 agcgcaacgc aattaatgtg agttacctca ctcattaggc accccaggct ttacacttta 7261 tgcttccggc tcctatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 7321 gctatgacca tgattacgcc aagcgcgcaa ttaacc //