LOCUS Exported 5983 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast expression vector containing HIS10-TEV-HA (HTH) epitope tag downstream of the GAL1 promoter. ACCESSION . VERSION . KEYWORDS pYES2-HTH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5983) AUTHORS Singh RK, Kabbaj MH, Paik J, Gunjan A TITLE Histone levels are regulated by phosphorylation and ubiquitylation-dependent proteolysis. JOURNAL Nat Cell Biol. 2009 Aug;11(8):925-33. doi: 10.1038/ncb1903. Epub 2009 Jul 5. PUBMED 19578373 REFERENCE 2 (bases 1 to 5983) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5983 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..442 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 1..118 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 413..436 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" primer_bind 474..493 /label=T7 /note="T7 promoter, forward primer" promoter 474..492 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 515..544 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 569..589 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 599..625 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" terminator 731..978 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(747..765) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(1055..1072) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1226..1814) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1306..1325) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1985..2845) /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 2608..2627 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS complement(2941..3744) /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(3745..3960) /gene="S. cerevisiae URA3" /label=URA3 promoter rep_origin 4561..5438 /label=2u ori /note="yeast 2u plasmid origin of replication" rep_origin complement(5449..5962) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5644..5665) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5856..5875 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc 61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa 121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc 181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat 241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt 301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg 361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc 421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgga ctactagcag ctgtaatacg 481 actcactata gggaatatta agcttgctag aatgcatcac catcatcacc atcatcacca 541 ccatgtggat tacgatatcc caactaccga aaacttgtac ttccaaggtg gcggccgcta 601 cccatacgat gttccagatt acgcccatgg taccgagctc ggatccacta gtaacggccg 661 ccagtgtgct ggaattctgc agatatccat cacactggcg gccgctcgag catgcatcta 721 gagggccgca tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc 781 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt 841 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac 901 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc 961 tcgaaggctt taatttgcaa gctgcggccc tgcattaatg aatcggccaa cgcgcgggga 1021 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 1081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 1141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 1201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 1261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 1321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 1381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 1441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 1501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 1561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 1621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 1681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 1741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 1801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 1861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 1921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 1981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 2041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 2101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 2161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 2221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 2281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 2341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 2401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 2461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 2521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 2581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 2641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 2701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 2761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 2821 cgacacggaa atgttgaata ctcatactct tcctttttca atgggtaata actgatataa 2881 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt 2941 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc 3001 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc 3061 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt 3121 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc 3181 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac 3241 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat 3301 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat 3361 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac 3421 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa 3481 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga 3541 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc 3601 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg 3661 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg 3721 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg 3781 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata 3841 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc 3901 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa 3961 aagctagctt atcgatgata agctgtcaaa catgagaatt aattccacgg actatagact 4021 atacctagta tactccgtct actgtacgat acacttccgc tcaggtcctt gtcctttaac 4081 gaggccttac cactcttttg ttactctatt gatccagctc agcaaaggca gtgtgatcta 4141 agattctatc ttcgcgatgt agtaaaacta gctagaccga gaaagagact agaaatgcaa 4201 aaggcacttc tacaatggct gccatcatta ttatccgatg tgacgctgca gcttctcaat 4261 gatattcgaa tacgctttga ggagatacag cctaatatcc gacaaactgt tttacagatt 4321 tacgatcgta cttgttaccc atcattgaat tttgaacatc cgaacctggg agttttccct 4381 gaaacagata gtatatttga acctgtataa taatatatag tctagcgctt tacggaagac 4441 aatgtatgta tttcggttcc tggagaaact attgcatcta ttgcataggt aatcttgcac 4501 gtcgcatccc cggttcattt tctgcgtttc catcttgcac ttcaatagca tatctttgtt 4561 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt 4621 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt 4681 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa 4741 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc 4801 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc 4861 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg 4921 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg 4981 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta 5041 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat 5101 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag 5161 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca 5221 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa 5281 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg 5341 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga 5401 tacttttgag caatgtttgt ggaagcggta ttcgcaatgg gaagctccac cccggttgat 5461 aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat tgtaagcgtt 5521 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 5581 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 5641 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 5701 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 5761 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 5821 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 5881 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 5941 aatgcgccgc tacagggcgc gtggggatcg atccactagt cca //