LOCUS       sequence-226342-        7340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synthetic acetylation substrate consisting of GFP fusion with 3 
            repeats of the Gcn5 consensus motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7340)
  AUTHORS   Rossl A, Denoncourt A, Lin MS, Downey M
  TITLE     A synthetic non-histone substrate to study substrate targeting by 
            the Gcn5 HAT and sirtuin HDACs.
  JOURNAL   J Biol Chem. 2019 Apr 19;294(16):6227-6239. doi: 
            10.1074/jbc.RA118.006051. Epub 2019 Feb 25.
  PUBMED    30804216
REFERENCE   2  (bases 1 to 7340)
  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.RA118.006051"; journalName: "J Biol Chem"; date: 
            "2019-04-19- 19"; volume: "294"; issue: "16"; pages: "6227-6239"
COMMENT     Sequence Label: GFP-X3
FEATURES             Location/Qualifiers
     source          1..7340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..30
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14..30
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             50..763
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(80..108)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     703..723
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(829..845)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(830..846)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      848..1095
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(864..882)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(1117..1135)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1156..1172)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1156..1172)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1169..1191)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1180..1196
                     /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)."
     promoter        complement(join(1204..1210,1211..1228,1229..1234))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1249..1270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1387..1404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1558..2146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1638..1657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2317..3108,3109..3177))
                     /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     2940..2959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3178..3282)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3319..3822
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3863..3881
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3919..3941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4041..4060
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4084..4299
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4300..5103
                     /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"
     rep_origin      complement(5234..5689)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5371..5392)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5583..5602
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5815..5837
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5829..5846
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5830..5846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5853..5872
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5853..5871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6636..7340
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 ccaagctggc cgctctagaa ctagtggatc ccccgggctg caggaattca tgagtaaagg
       61 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
      121 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
      181 taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt
      241 cacttatggt gttcaatgct tttcaagata cccagatcat atgaaacggc atgacttttt
      301 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      361 gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
      421 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
      481 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa
      541 cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca
      601 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca
      661 atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt
      721 aacagctgct gggattacac atggcatgga tgaactatac aaaggttctt caaaaagacc
      781 tggatcatct aagaggcctg gctcatcaaa gagaccataa aagcttatcg ataccgtcga
      841 cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct
      901 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
      961 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     1021 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     1081 aaggctttaa tttgcggccg gtaccagctt ttgttccctt tagtgagggt taatttcgag
     1141 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1201 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1261 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1321 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3301 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     3361 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     3421 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     3481 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     3541 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     3601 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     3661 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     3721 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     3781 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtctaag aaaccattat
     3841 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     3901 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3961 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4021 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataccac
     4081 cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc ttgaaatttt
     4141 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     4201 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     4261 ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc tacatataag
     4321 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     4381 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     4441 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     4501 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     4561 ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     4621 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     4681 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     4741 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     4801 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     4861 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     4921 gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga
     4981 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     5041 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     5101 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     5161 taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5221 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5281 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5341 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5401 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5461 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5521 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5581 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5641 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     5701 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5761 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     5821 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
     5881 agctcgccgg gatcgaagaa atgatggtaa atgaaatagg aaatcaagga gcatgaaggc
     5941 aaaagacaaa tataagggtc gaacgaaaaa taaagtgaaa agtgttgata tgatgtattt
     6001 ggctttgcgg cgccgaaaaa acgagtttac gcaattgcac aatcatgctg actctgtggc
     6061 ggacccgcgc tcttgccggc ccggcgataa cgctgggcgt gaggctgtgc ccggcggagt
     6121 tttttgcgcc tgcattttcc aaggtttacc ctgcgctaag gggcgagatt ggagaagcaa
     6181 taagaatgcc ggttggggtt gcgatgatga cgaccacgac aactggtgtc attatttaag
     6241 ttgccgaaag aacctgagtg catttgcaac atgagtatac tagaagaatg agccaagact
     6301 tgcgagacgc gagtttgccg gtggtgcgaa caatagagcg accatgacct tgaaggtgag
     6361 acgcgcataa ccgctagagt actttgaaga ggaaacagca atagggttgc taccagtata
     6421 aatagacagg tacatacaac actggaaatg gttgtctgtt tgagtacgct ttcaattcat
     6481 ttgggtgtgc actttattat gttacaatat ggaagggaac tttacacttc tcctatgcac
     6541 atatattaat taaagtccaa tgctagtaga gaaggggggt aacacccctc cgcgctcttt
     6601 tccgattttt ttctaaaccg tggaatattt cggatatcct tttgttgttt ccgggtgtac
     6661 aatatggact tcctcttttc tggcaaccaa acccatacat cgggattcct ataatacctt
     6721 cgttggtctc cctaacatgt aggtggcgga ggggagatat acaatagaac agataccaga
     6781 caagacataa tgggctaaac aagactacac caattacact gcctcattga tggtggtaca
     6841 taacgaacta atactgtagc cctagacttg atagccatca tcatatcgaa gtttcactac
     6901 cctttttcca tttgccatct attgaagtaa taataggcgc atgcaacttc ttttcttttt
     6961 ttttcttttc tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg
     7021 gaagacacta aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag
     7081 agctgatgag gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact
     7141 actctctaat gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa
     7201 aaagtttgct gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc
     7261 tcgtcattgt tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta
     7321 taccaagcat acaatcaact
//
