LOCUS Exported 9186 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTH727-CEN-RLuc/staCFLuc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9186) AUTHORS Chu D, Kazana E, Bellanger N, Singh T, Tuite MF, von der Haar T TITLE Translation elongation can control translation initiation on eukaryotic mRNAs. JOURNAL EMBO J. 2014 Jan 1;33(1):21-34. doi: 10.1002/embj.201385651. Epub 2013 Dec 19. PUBMED 24357599 REFERENCE 2 (bases 1 to 9186) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9186 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 70..573 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 610..714 /gene="bla" /label=AmpR promoter CDS 715..1575 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(933..952) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1746..2334 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2488..2505 /label=L4440 /note="L4440 vector, forward primer" promoter 2658..2688 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2696..2712 /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 2701..2723 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2720..2736 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2720..2736 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 2757..2775 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" terminator 2824..3123 /label=ADH2 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 2 (ADH2) gene" CDS complement(3154..4086) /codon_start=1 /product="luciferase from the anthozoan coelenterate Renilla reniformis (sea pansy)" /label=Rluc /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" promoter 4499..5143 /gene="S. cerevisiae TDH3" /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase" primer_bind 5036..5059 /label=GPDpro-F /note="S. cerevisiae GPD promoter, forward primer" primer_bind complement(5217..5235) /label=LucNrev /note="Firefly luciferase, reverse primer" terminator 6950..7137 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(7005..7029) /label=pHybLex-R /note="ADH terminator, reverse primer" primer_bind complement(7202..7221) /label=T7 /note="T7 promoter, forward primer" promoter complement(7203..7221) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7228..7245) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7228..7244) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7237..7259) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7389..7844 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7476..7495) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7686..7707 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(7973..8776) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(8777..8992) /gene="S. cerevisiae URA3" /label=URA3 promoter primer_bind complement(9017..9036) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9136..9158 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata 121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt 181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa 241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata 301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct 361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata 421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact 481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa 541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg 601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt 1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc 2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc 2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact 2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg 2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt 2761 aaccctcact aaagggaaca aaagctgggt accgggcccc ccctcgaggg catgcgaagg 2821 aaaatgagaa atatcgaggg agacgattca gaggagcagg acaaactata accgactgtt 2881 tgttggagga tgccgtacat aacgaacact gctgaagcta ccatgtctac agtttagagg 2941 aatgggtaca actcacaggc gagggatggt gttcactcgt gctagcaaac gcggtgggag 3001 caaaaagtag aatattatct tttattcgtg aaacttcgaa cactgtcatc taaagatgct 3061 atatactaat ataggcatac ttgataatgg aaactataaa tcgtgaaggc ataggagatc 3121 cgcttattta gaagtgtcga attcttacga catttgttca tttttgagaa ctcgctcaac 3181 gaacgatttg atatattttc ccatttcatc aggtgcatct tcttgcgaaa aatgaagacc 3241 ttttactttg acaaattcag tattaggaaa cttcttggca ccttcaacaa tagcattgga 3301 aaagaatcct gggtccgatt caataaacat ttttggtaaa tcatcacttg cacgtagata 3361 agcattataa ttcctaacaa tttgtacaac gtcaggttta ccacctttta ctaacgggat 3421 ttcacgaggc catgataatg ttggacgacg aacttcacct ttctctttga atggttcaag 3481 atatgctgca aattcttctg gttctaactt tctcatgatt tttgatggca acatggtttc 3541 cacgaagaag ttattctcca aaaccatttt ttctccttct tcagatttga tcaacgcaat 3601 atcttcttca atatcaggcc attcatccca tgattcaatc acatctacta cactttcagc 3661 gtgaactatt gctttgatct tatcttgatg ctcatagcta taatgaaatg ccaaacaagc 3721 accccaatca tggccgacaa aaatgatctt ctttggtaaa ttaagaagtt caaaccatgc 3781 agtaagatat ttgtaatgat caagtaacct ataagaacca ttaccagatt tgcctgattt 3841 gcccatacca ataaggtctg gtataataca ccgcgctact ggctcaatat gtggcacaac 3901 atgtcgccat aaataagaag aggccgcgtt accatgtaaa aaaataacag cattttctgc 3961 atgtttttct gaatcataat aattaataaa tgaatcaaga acattcattt gtttacatct 4021 ggcccaccac tgcggaccag ttatcatccg tttcctttgt tctggatcat aaactttcga 4081 agtcatcccg gggagttgat tgtatgcttg gtatagcttg aaatattgtg cagaaaaaga 4141 aacaaggaag aaagggaacg agaacaatga cgaggaaaca aaagattaat aattgcaggt 4201 ctatttatac ttgatagcaa agcggcaaac tttttttatt tcaaattcaa gtaactggaa 4261 ggaaggccgt ataccgttgc tcattagaga gtagtgtgcg tgaatgaagg aaggaaaaag 4321 tttcgtgtgt tcgaagatac ccctcatcag ctctggaaca acgacatctg ttggtgctgt 4381 ctttgtcgtt aattttttcc tttagtgtct tccatcattt tttttgtcat tgcggatatg 4441 gtgagacaac aacgggggag agagaaaaga aaaaaaaaga aaagaatccg ggagtttatc 4501 attatcaata ctgccatttc aaagaatacg taaataatta atagtagtga ttttcctaac 4561 tttatttagt caaaaaatta gccttttaat tctgctgtaa cccgtacatg cccaaaatag 4621 ggggcgggtt acacagaata tataacatcg taggtgtctg ggtgaacagt ttattcctgg 4681 catccactaa atataatgga gcccgctttt taagctggca tccagaaaaa aaaagaatcc 4741 cagcaccaaa atattgtttt cttcaccaac catcagttca taggtccatt ctcttagcgc 4801 aactacagag aacaggggca caaacaggca aaaaacgggc acaacctcaa tggagtgatg 4861 caacctgcct ggagtaaatg atgacacaag gcaattgacc cacgcatgta tctatctcat 4921 tttcttacac cttctattac cttctgctct ctctgatttg gaaaaagctg aaaaaaaagg 4981 ttgaaaccag ttccctgaaa ttattcccct acttgactaa taagtatata aagaacggta 5041 ggtattgatt gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt 5101 tagtcttttt tttagtttta aaacaccaag aacttagttt cgacggatcc atggaagacg 5161 ccaaaaacat aaagaaaggc ccggcgccat tctatcctct agaggatgga accgctggag 5221 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag 5281 atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc gttcggttgg 5341 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa 5401 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc 5461 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgaacatt tcgcagccta 5521 ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaaattac 5581 caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga 5641 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtaccag 5701 agtcctttga tcgtgacaaa acaattgcac tgataatgaa ttcctctgga tctactgggt 5761 tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg catgccagag 5821 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc 5881 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct 5941 taatgtatag atttgaagaa gagctgtttt tacgatccct tcaggattac aaaattcaaa 6001 gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg attgacaaat 6061 acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg aaagaagtcg 6121 gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat gggctcactg 6181 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta 6241 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg 6301 ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt tatgtaaaca 6361 atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag 6421 cttactggga cgaagacgaa cacttcttca tagttgaccg cttgaagtct ttaattaaat 6481 acaaaggata tcaggtggcc cccgctgaat tggaatcgat attgttacaa caccccaaca 6541 tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg 6601 ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca 6661 gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga 6721 aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga 6781 agggcggaaa gtaagtcgac ctgcaggcat gcaagctttg gacttcttcg ccagaggttt 6841 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat 6901 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt 6961 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta 7021 aagtgactct taggttttaa aacgaaaatt cttgttcttg agtaactctt tcctgtaggt 7081 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat 7141 gccggtcgaa attcccctac cctatgaaaa gcggccgcca ccgcggtgga gctccaattc 7201 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga 7261 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccccttcg ccagctggcg 7321 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 7381 atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 7441 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 7501 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 7561 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 7621 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 7681 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 7741 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 7801 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat gcggtatttt 7861 ctccttacgc atctgtgcgg tatttcacac cgcagggtaa taactgatat aattaaattg 7921 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt 7981 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta 8041 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg 8101 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta 8161 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc 8221 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct 8281 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc 8341 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc 8401 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag 8461 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa 8521 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag 8581 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact 8641 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt 8701 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat 8761 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg 8821 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc 8881 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat 8941 ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aattgaaaag 9001 cgtggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc 9061 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac 9121 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac 9181 gcgcga //