LOCUS       sequence-226330-        7304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP tethered to a single repeat of the Gcn5 consensus motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7304)
  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 7304)
  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-X1
FEATURES             Location/Qualifiers
     source          1..7304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..705
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     719..735
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     719..735
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             755..1468
                     /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(785..813)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1408..1428
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1498..1514)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1499..1515)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1517..1764
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1533..1551)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(1786..1804)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1825..1841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1825..1841)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1838..1860)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1849..1865
                     /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(1873..1879,1880..1897,1898..1903))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1918..1939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2056..2073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2227..2815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2307..2326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2986..3777,3778..3846))
                     /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     3609..3628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3847..3951)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3988..4491
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4532..4550
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4588..4610)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4710..4729
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4753..4968
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4969..5772
                     /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(5903..6358)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6040..6061)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6252..6271
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6484..6506
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6498..6515
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6499..6515
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6522..6541
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6522..6540
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atccttttgt tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca
       61 tacatcggga ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga
      121 gatatacaat agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt
      181 acactgcctc attgatggtg gtacataacg aactaatact gtagccctag acttgatagc
      241 catcatcata tcgaagtttc actacccttt ttccatttgc catctattga agtaataata
      301 ggcgcatgca acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca
      361 tatccgcaat gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca
      421 gcaccaacag atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt
      481 tttccttcct tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc
      541 cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac
      601 ctgcaattat taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc
      661 tttttctgca caatatttca agctatacca agcatacaat caactccaag ctggccgctc
      721 tagaactagt ggatcccccg ggctgcagga attcatgagt aaaggagaag aacttttcac
      781 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      841 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
      901 tactggaaaa ctacctgttc catggccaac acttgtcact actttcactt atggtgttca
      961 atgcttttca agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc
     1021 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
     1081 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     1141 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     1201 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
     1261 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     1321 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     1381 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     1441 tacacatggc atggatgaac tatacaaagg ttcttcaaaa agaccttaaa agcttatcga
     1501 taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
     1561 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     1621 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     1681 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
     1741 ggacgctcga aggctttaat ttgcggccgg taccagcttt tgttcccttt agtgagggtt
     1801 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1861 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1921 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1981 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2041 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2101 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2161 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2221 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2281 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2341 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2401 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2461 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2521 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2581 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2641 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2701 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2761 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2821 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2881 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2941 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3601 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3841 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3901 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3961 ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat
     4021 tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat
     4081 agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt
     4141 tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga
     4201 agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc
     4261 tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat
     4321 tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt
     4381 actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat
     4441 tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga
     4501 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4561 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4621 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4681 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4741 ccataccacc ttttcaattc atcatttttt ttttattctt ttttttgatt tcggtttcct
     4801 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     4861 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     4921 ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat gtcgaaagct
     4981 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     5041 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     5101 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     5161 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     5221 aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt caaattgcag
     5281 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     5341 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     5401 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     5461 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     5521 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     5581 ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga tgatgtggtc
     5641 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     5701 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     5761 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     5821 gcttcaattt aattatatca gttattaccc tatgcggtgt gaaataccgc acagatgcgt
     5881 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     5941 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     6001 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     6061 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     6121 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     6181 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     6241 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     6301 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     6361 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6421 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6481 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
     6541 gcgaattgga gctcgccggg atcgaagaaa tgatggtaaa tgaaatagga aatcaaggag
     6601 catgaaggca aaagacaaat ataagggtcg aacgaaaaat aaagtgaaaa gtgttgatat
     6661 gatgtatttg gctttgcggc gccgaaaaaa cgagtttacg caattgcaca atcatgctga
     6721 ctctgtggcg gacccgcgct cttgccggcc cggcgataac gctgggcgtg aggctgtgcc
     6781 cggcggagtt ttttgcgcct gcattttcca aggtttaccc tgcgctaagg ggcgagattg
     6841 gagaagcaat aagaatgccg gttggggttg cgatgatgac gaccacgaca actggtgtca
     6901 ttatttaagt tgccgaaaga acctgagtgc atttgcaaca tgagtatact agaagaatga
     6961 gccaagactt gcgagacgcg agtttgccgg tggtgcgaac aatagagcga ccatgacctt
     7021 gaaggtgaga cgcgcataac cgctagagta ctttgaagag gaaacagcaa tagggttgct
     7081 accagtataa atagacaggt acatacaaca ctggaaatgg ttgtctgttt gagtacgctt
     7141 tcaattcatt tgggtgtgca ctttattatg ttacaatatg gaagggaact ttacacttct
     7201 cctatgcaca tatattaatt aaagtccaat gctagtagag aaggggggta acacccctcc
     7261 gcgctctttt ccgatttttt tctaaaccgt ggaatatttc ggat
//
