LOCUS       sequence-226511-        7348 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GFP-X2 where both of the consensus motif's lysines are mutated to 
            arginine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7348)
  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 7348)
  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-X2DM
FEATURES             Location/Qualifiers
     source          1..7348
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    37..540
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     581..599
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(637..659)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     759..778
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        802..1017
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1018..1821
                     /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(1952..2407)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2089..2110)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2301..2320
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2555
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2547..2564
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2548..2564
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2571..2590
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2571..2589
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3354..4058
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     4072..4088
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4072..4088
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4108..4821
                     /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(4138..4166)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4761..4781
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4869..4885)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4870..4886)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4888..5135
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4904..4922)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(5157..5175)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5196..5212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5196..5212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5209..5231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5220..5236
                     /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(5244..5250,5251..5268,5269..5274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5427..5444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5598..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5678..5697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6357..7148,7149..7217))
                     /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     6980..6999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7218..7322)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat
       61 aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa
      121 agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat
      181 actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt
      241 ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga
      301 atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt
      361 tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct
      421 tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa
      481 ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc
      541 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      601 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      661 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      721 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      781 gagagtgcac cataccacct tttcaattca tcattttttt tttattcttt tttttgattt
      841 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
      901 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
      961 ccagtattct taacccaact gcacagaaca aaaacctgca ggaaacgaag ataaatcatg
     1021 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     1081 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     1141 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     1201 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     1261 gccaagtaca attttttact cttcgaagac agaaaatttg ctgacattgg taatacagtc
     1321 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     1381 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     1441 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     1501 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     1561 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     1621 cccggtgtgg gtttagatga caagggagac gcattgggtc aacagtatag aaccgtggat
     1681 gatgtggtct ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     1741 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     1801 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     1861 caaattagag cttcaattta attatatcag ttattaccct atgcggtgtg aaataccgca
     1921 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     1981 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2041 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2101 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2161 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2221 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2281 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2341 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2401 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2461 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2521 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
     2581 cactataggg cgaattggag ctcgccggga tcgaagaaat gatggtaaat gaaataggaa
     2641 atcaaggagc atgaaggcaa aagacaaata taagggtcga acgaaaaata aagtgaaaag
     2701 tgttgatatg atgtatttgg ctttgcggcg ccgaaaaaac gagtttacgc aattgcacaa
     2761 tcatgctgac tctgtggcgg acccgcgctc ttgccggccc ggcgataacg ctgggcgtga
     2821 ggctgtgccc ggcggagttt tttgcgcctg cattttccaa ggtttaccct gcgctaaggg
     2881 gcgagattgg agaagcaata agaatgccgg ttggggttgc gatgatgacg accacgacaa
     2941 ctggtgtcat tatttaagtt gccgaaagaa cctgagtgca tttgcaacat gagtatacta
     3001 gaagaatgag ccaagacttg cgagacgcga gtttgccggt ggtgcgaaca atagagcgac
     3061 catgaccttg aaggtgagac gcgcataacc gctagagtac tttgaagagg aaacagcaat
     3121 agggttgcta ccagtataaa tagacaggta catacaacac tggaaatggt tgtctgtttg
     3181 agtacgcttt caattcattt gggtgtgcac tttattatgt tacaatatgg aagggaactt
     3241 tacacttctc ctatgcacat atattaatta aagtccaatg ctagtagaga aggggggtaa
     3301 cacccctccg cgctcttttc cgattttttt ctaaaccgtg gaatatttcg gatatccttt
     3361 tgttgtttcc gggtgtacaa tatggacttc ctcttttctg gcaaccaaac ccatacatcg
     3421 ggattcctat aataccttcg ttggtctccc taacatgtag gtggcggagg ggagatatac
     3481 aatagaacag ataccagaca agacataatg ggctaaacaa gactacacca attacactgc
     3541 ctcattgatg gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc
     3601 atatcgaagt ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat
     3661 gcaacttctt ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc
     3721 aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa
     3781 cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt
     3841 ccttcattca cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac
     3901 ttgaatttga aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgcaat
     3961 tattaatctt ttgtttcctc gtcattgttc tcgttccctt tcttccttgt ttctttttct
     4021 gcacaatatt tcaagctata ccaagcatac aatcaactcc aagctggccg ctctagaact
     4081 agtggatccc ccgggctgca ggaattcatg agtaaaggag aagaactttt cactggagtt
     4141 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     4201 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga
     4261 aaactacctg ttccatggcc aacacttgtc actactttca cttatggtgt tcaatgcttt
     4321 tcaagatacc cagatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt
     4381 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa
     4441 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     4501 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac
     4561 atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt
     4621 gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     4681 cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc
     4741 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     4801 ggcatggatg aactatacaa aggttcttca agaagacctg gatcatctag gaggccttaa
     4861 aagcttatcg ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac
     4921 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     4981 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     5041 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     5101 agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtaccagctt ttgttccctt
     5161 tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5221 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5281 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5401 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6001 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7321 cgcacatttc cccgaaaagt gccacctg
//
