LOCUS Exported 6649 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION overexpression of HST2 in S. cerevisiae, HIS3 marker. ACCESSION . VERSION . KEYWORDS p413TEF-HST2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6649) AUTHORS Ralser M, Zeidler U, Lehrach H TITLE Interfering with glycolysis causes Sir2-dependent hyper-recombination of Saccharomyces cerevisiae plasmids. JOURNAL PLoS One. 2009;4(4):e5376. doi: 10.1371/journal.pone.0005376. Epub 2009 Apr 24. PUBMED 19390637 REFERENCE 2 (bases 1 to 6649) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6649 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(9..28) /label=T7 /note="T7 promoter, forward primer" promoter complement(10..28) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(38..55) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(38..54) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(47..69) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 199..654 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(286..305) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 496..517 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(914..1573) /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(1574..1760) /gene="S. cerevisiae HIS3" /label=HIS3 promoter primer_bind complement(1907..1926) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2026..2048 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2086..2104) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 2146..2649 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 2686..2790 /gene="bla" /label=AmpR promoter CDS 2791..3651 /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(3009..3028) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3822..4410 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4311..4330 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4564..4581 /label=L4440 /note="L4440 vector, forward primer" promoter 4734..4764 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4772..4788 /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 4777..4799 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4796..4812 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4796..4812 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 4831..4851 /label=T3 /note="T3 promoter, forward primer" promoter 4833..4851 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 4869..5267 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind 5271..5287 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 5271..5287 /label=pBluescriptSK /note="For pBluescript vector" terminator 6393..6640 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6409..6427) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" ORIGIN 1 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc 61 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 121 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 181 tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 241 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 301 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 361 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 421 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 481 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 541 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 601 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat 661 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagatc cgtcgagttc 721 aagagaaaaa aaaagaaaaa gcaaaaagaa aaaaggaaag cgcgcctcgt tcagaatgac 781 acgtatagaa tgatgcatta ccttgtcatc ttcagtatca tactgttcgt atacatactt 841 actgacattc ataggtatac atatatacac atgtatatat atcgtatgct gcagctttaa 901 ataatcggtg tcactacata agaacacctt tggtggaggg aacatcgttg gtaccattgg 961 gcgaggtggc ttctcttatg gcaaccgcaa gagccttgaa cgcactctca ctacggtgat 1021 gatcattctt gcctcgcaga caatcaacgt ggagggtaat tctgctagcc tctgcaaagc 1081 tttcaagaaa atgcgggatc atctcgcaag agagatctcc tactttctcc ctttgcaaac 1141 caagttcgac aactgcgtac ggcctgttcg aaagatctac caccgctctg gaaagtgcct 1201 catccaaagg cgcaaatcct gatccaaacc tttttactcc acgcgccagt agggcctctt 1261 taaaagcttg accgagagca atcccgcagt cttcagtggt gtgatggtcg tctatgtgta 1321 agtcaccaat gcactcaacg attagcgacc agccggaatg cttggccaga gcatgtatca 1381 tatggtccag aaaccctata cctgtgtgga cgttaatcac ttgcgattgt gtggcctgtt 1441 ctgctactgc ttctgcctct ttttctggga agatcgagtg ctctatcgct aggggaccac 1501 cctttaaaga gatcgcaatc tgaatcttgg tttcatttgt aatacgcttt actagggctt 1561 tctgctctgt catctttgcc ttcgtttatc ttgcctgctc attttttagt atattcttcg 1621 aagaaatcac attactttat ataatgtata attcattatg tgataatgcc aatcgctaag 1681 aaaaaaaaag agtcatccgc taggggaaaa aaaaaaatga aaatcattac cgaggcataa 1741 aaaaatatag agtgtactag aggaggccaa gagtaataga aaaagaaaat tgcgggaaag 1801 gactgtgtta tgacttccct gactaatgcc gtgttcaaac gatacctggc agtgactcct 1861 agcgctcacc aagctcttaa aacgggaatt tatggtgcac tctcagtaca atctgctctg 1921 atgccgcata gttaagccag ccccgacacc cgccaacacg cgctgacgcg ccctgacggg 1981 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 2041 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc 2101 tatttttata ggttaatgtc atgataataa tggtttctta ggacggatcg cttgcctgta 2161 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta 2221 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac 2281 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac 2341 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag 2401 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat 2461 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc 2521 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt 2581 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata 2641 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 2701 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 2761 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 2821 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 2881 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 2941 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 3001 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 3061 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 3121 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 3181 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 3241 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 3301 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 3361 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 3421 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 3481 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 3541 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 3601 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 3661 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 3721 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 3781 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 3841 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 3901 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 3961 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 4021 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 4081 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 4141 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 4201 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 4261 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 4321 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 4381 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 4441 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 4501 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 4561 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 4621 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc 4681 agtgagcgca acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac 4741 tttatgcttc cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga 4801 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag 4861 ctggagctca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga 4921 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct 4981 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac 5041 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt 5101 cttgaaaatt ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt 5161 aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt 5221 ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt tctagaacta 5281 gtggatcccc cgggctgcag gaattcatgt ctgtttctac cgcctctaca gagatgtcgg 5341 ttaggaagat cgctgctcat atgaaaagca accccaacgc taaagtaatc tttatggtgg 5401 gtgccgggat atccacttct tgtgggatac cagacttccg atctccggga actggcctgt 5461 accataactt ggccaggttg aagttgccgt accctgaagc agtgttcgat gtcgattttt 5521 tccaatcgga tcctttgcca ttttacactt tggctaaaga gttgtatccc ggaaatttca 5581 gaccttctaa gtttcactat cttttgaaat tgtttcaaga caaagacgtt ttgaagagag 5641 tatataccca gaatatagac actttagaaa gacaggccgg tgttaaggat gatctgatca 5701 tcgaggccca tggcagtttt gctcactgtc attgtatcgg atgtggtaaa gtttatcccc 5761 cacaggtttt caaatccaag ctggcagagc atcctataaa ggattttgtc aagtgtgatg 5821 tgtgcggtga actggtaaag ccggcaatag ttttttttgg cgaagatcta ccggattcct 5881 tttcggaaac ttggttaaac gattcagagt ggcttcgtga aaaaatcacg acatctggaa 5941 aacatcctca gcagcctctt gtcatcgtcg ttggcacctc actagccgtt tatccctttg 6001 cgtctctgcc cgaagaaatt cctcgaaagg taaaaagagt tctgtgcaat ttggaaacgg 6061 tgggcgattt taaagctaac aagaggccta cagatttaat agttcatcag tactcggatg 6121 agtttgccga gcaactcgtt gaagaattag gttggcaaga agatttcgaa aagatactta 6181 cggcccaagg tggaatgggg gataactcta aagagcaatt acttgaaatt gtacatgatt 6241 tagagaattt gagtttggat caatctgaac atgagagtgc agacaagaag gataagaagc 6301 tacagcgtct caatggccat gacagcgatg aagatggagc aagcaactcc tcttcttcac 6361 aaaaagccgc taaagaataa gtcgacctcg agtcatgtaa ttagttatgt cacgcttaca 6421 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt 6481 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca 6541 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt 6601 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtac //