LOCUS       sequence-170869-        6363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of HHT2 with His10-TEV-HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6363)
  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 6363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb1903"; 
            journalName: "Nat Cell Biol"; date: "2009-08"; volume: "11"; issue: 
            "8"; pages: "925-33"
COMMENT     Sequence Label: pYES2-HTH-HHT2
FEATURES             Location/Qualifiers
     source          1..6363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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      1111..1358
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1127..1145)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1435..1452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1606..2194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1686..1705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2365..3156,3157..3225))
                     /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     2988..3007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3321..4124)
                     /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(4125..4340)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      4941..5818
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5829..6342)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6024..6045)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6236..6255
                     /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 ggaattctgg atccaatggc cagaactaaa caaacagcta gaaaatccac
      721 tggtggtaaa gccccaagaa aacaattagc ctccaaggct gccagaaaat ccgccccatc
      781 taccggtggt gttaagaagc ctcacagata taagccaggt actgttgcct tgagagaaat
      841 tagaagattc caaaaatcta ctgaactgtt gatcagaaag ttacctttcc aaagattggt
      901 cagagaaatc gctcaagatt tcaagaccga cttgagattt caatcttctg ctatcggtgc
      961 tttgcaagaa tccgtcgaag catacttagt ctctttgttt gaagacacta atctggctgc
     1021 tattcacgct aagcgtgtta ctatccaaaa gaaggatatc aaattggcca gaagactaag
     1081 aggtgaaaga tcatgatcta gagggccgca tcatgtaatt agttatgtca cgcttacatt
     1141 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     1201 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     1261 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     1321 ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa gctgcggccc tgcattaatg
     1381 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1441 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1501 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1561 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1621 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1681 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1741 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1801 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1861 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1921 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1981 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2041 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2101 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2161 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2221 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2281 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2341 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2401 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2461 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2521 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2581 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2641 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2701 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2761 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2821 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2881 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2941 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3001 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3061 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3121 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3181 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3241 atgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     3301 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     3361 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     3421 cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt tctatactgt
     3481 tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg
     3541 taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg
     3601 tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg
     3661 catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc
     3721 tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc
     3781 cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca
     3841 gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc
     3901 ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa
     3961 attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat
     4021 gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga
     4081 ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc
     4141 ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca
     4201 acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     4261 tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa
     4321 aaaaaaaatg atgaattgaa aagctagctt atcgatgata agctgtcaaa catgagaatt
     4381 aattccacgg actatagact atacctagta tactccgtct actgtacgat acacttccgc
     4441 tcaggtcctt gtcctttaac gaggccttac cactcttttg ttactctatt gatccagctc
     4501 agcaaaggca gtgtgatcta agattctatc ttcgcgatgt agtaaaacta gctagaccga
     4561 gaaagagact agaaatgcaa aaggcacttc tacaatggct gccatcatta ttatccgatg
     4621 tgacgctgca gcttctcaat gatattcgaa tacgctttga ggagatacag cctaatatcc
     4681 gacaaactgt tttacagatt tacgatcgta cttgttaccc atcattgaat tttgaacatc
     4741 cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa taatatatag
     4801 tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact attgcatcta
     4861 ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac
     4921 ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
     4981 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     5041 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
     5101 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     5161 gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta
     5221 caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct
     5281 cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg
     5341 ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt
     5401 cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc
     5461 cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt
     5521 gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact
     5581 acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta
     5641 ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga
     5701 gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac
     5761 agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatgg
     5821 gaagctccac cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa
     5881 atatttaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5941 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     6001 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     6061 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6121 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6181 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6241 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6301 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatcg atccactagt
     6361 cca
//
