LOCUS       sequence-164029-        5933 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  yeast expression of EEF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5933)
  AUTHORS   Davydova E, Ho AY, Malecki J, Moen A, Enserink JM, Jakobsson ME, 
            Loenarz C, Falnes PO
  TITLE     Identification and characterization of a novel evolutionarily 
            conserved lysine-specific methyltransferase targeting eukaryotic 
            translation elongation factor 2 (eEF2).
  JOURNAL   J Biol Chem. 2014 Oct 31;289(44):30499-510. doi: 
            10.1074/jbc.M114.601658. Epub 2014 Sep 17.
  PUBMED    25231979
REFERENCE   2  (bases 1 to 5933)
  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: 
            "10.1074/jbc.M114.601658"; journalName: "J Biol Chem"; date: 
            "2014-10-31- 31"; volume: "289"; issue: "44"; pages: "30499-510"
COMMENT     Sequence Label: pYES260-sc.eEF2
FEATURES             Location/Qualifiers
     source          1..5933
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(5..252)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     218..236
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(383..403)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(410..427)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(443..462)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(444..462)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(494..936)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(500..523)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    819..936
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      955..1410
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1042..1061)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1479..2359
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        2958..3178
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3179..3982
                     /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"
     CDS             join(4078..4146,4147..4938)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5109..5697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5598..5617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5851..5868
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccgcaaat taaagccttc gagcgtccca aaaccttctc aagcaaggtt ttcagtataa
       61 tgttacatgc gtacacgcgt ctgtacagaa aaaaaagaaa aatttgaaat ataaataacg
      121 ttcttaatac taacataact ataaaaaaat aaatagggac ctagacttca ggttgtctaa
      181 ctccttcctt ttcggttaga gcggatgtgg ggggagggcg tgaatgtaag cgtgacataa
      241 ctaattacat gatgcggccc tctagatgca tgctcgagcg gccgccagtg tgatggatat
      301 ctgcagaatt ccagcacact ggcggccgtt actagtggat ccccgaccca tttgctgtcc
      361 accagtcatg ctagaccatg gcgccctgaa aataaagatt ctcgctagca tggtgatggt
      421 gatggtgcat aagcttaata ttccctatag tgagtcgtat tacagctgct agtagtccga
      481 tccggggttt tttctccttg acgttaaagt atagaggtat attaacaatt ttttgttgat
      541 acttttatta catttgaata agaagtaata caaaccgaaa atgttgaaag tattagttaa
      601 agtggttaat gcagtttttg catttatata tctgttaata gatcaaaaat catcgcttcg
      661 ctgattaatt accccagaaa taaggctaaa aaactaatcg cattatcatc ctatggttgt
      721 taatttgatt cgttcatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc
      781 ataaccataa aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca
      841 catctgcgtt tcaggaacgc gaccggtgag gacgaggacg cacggaggag agtcttcctt
      901 cggagggctg tcacccgctc ggcggcttct aatccgtact agtggatcat ccccacgcgc
      961 cctgtagcgc cccattaagc gcggcgggtg tggtggttac gcccagcgtg acccctacac
     1021 ttcccaccgc cctagccccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1081 ccggctttcc ccgtcaagct ctaaatgggg gcatccgttt acccttccga tttactgctt
     1141 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1201 cctgatagac cctttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1261 tgttggaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1321 ttttgccgat ttcgggctat tcgttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1381 attttaacaa aatattaacg tttacaattt aaatatttgc ttatacaatc ttcctgtttt
     1441 tggggctttt ctgattatca accggggtgg agcttcccat tgcgaatacc gcttccacaa
     1501 acattgctca aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac
     1561 ccatccacct ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt
     1621 atctctagta ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc
     1681 gaaaacaata cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga
     1741 aaccgttcat aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa
     1801 agtatgcgca atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat
     1861 gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg
     1921 gaagagaaaa tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa
     1981 agttatcaag agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct
     2041 ttgttagaaa aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt
     2101 ctttgttggt aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga
     2161 ttctttgttt gaaaaattag cgctctcgtc gcgttgcatt tttgttttac aaaaatgaag
     2221 cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc
     2281 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat
     2341 gaagcacaga tgcttcgtta acaaagatat gctattgaag tgcaagatgg aaacgcagaa
     2401 aatgaaccgg ggatgcgacg tgcaagatta cctatgcaat agatgcaata gtttctccag
     2461 gaaccgaaat acatacattg tcttccgtaa agcgctagac tatatattat tatacaggtt
     2521 caaatatact atctgtttca gggaaaactc ccaggttcgg atgttcaaaa ttcaatgatg
     2581 ggtaacaagt acgatcgtaa atctgtaaaa cagtttgtcg gatattaggc tgtatctcct
     2641 caaagcgtat tcgaatatca ttgagaagct gcagcgtcac atcggataat aatgatggca
     2701 gccattgtag aagtgccttt tgcatttcta gtctctttct cggtctagct agttttacta
     2761 catcgcgaag atagaatctt agatcacact gcctttgctg agctggatca atagagtaac
     2821 aaaagagtgg taaggcctcg ttaaaggaca aggacctgag cggaagtgta tcgtacagta
     2881 gacggagtat ctagtatagt ctatagtccg tggaattaat tctcatcttt gacagcttat
     2941 catcgataag ctagcttttc aattcaattc atcatttttt ttttattctt ttttttgatt
     3001 tcggtttctt tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
     3061 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
     3121 cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat
     3181 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
     3241 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
     3301 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
     3361 tgtggatatc ttgactgatt tttcaatgga gggcacagtt aagccgctaa aggcattatc
     3421 cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt
     3481 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
     3541 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
     3601 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     3661 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     3721 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     3781 acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga
     3841 tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     3901 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     3961 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     4021 acaaattaga gcttcaattt aattatatca gttattaccc attgaaaaag gaagagtatg
     4081 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4141 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4201 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4261 gaacgttttc caatgatgag cacttttaaa gttctgctat gtgatacact attatcccgt
     4321 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4381 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4441 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4501 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4561 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agagtgacac cacgatgcct
     4621 gtagcaatgc caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4681 cggcaacaat taatagactg aatggaggcg gataaagttg caggaccact tctgcgctcg
     4741 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4801 ggtatcattg cagcactggg gccagatggt aagcgctccc gtatcgtagt tatctacacg
     4861 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4921 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4981 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5041 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5101 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5161 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5221 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5281 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5341 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5401 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5461 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5521 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5581 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5641 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5701 gccagcaacg cggccttttt acggttcctg ggcttttgct ggccttttgc tcacatgttc
     5761 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5821 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5881 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
