LOCUS       sequence-226337-        7322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-X2 where the first consensus motif's lysine is mutated to 
            arginine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7322)
  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 7322)
  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-X2R1
FEATURES             Location/Qualifiers
     source          1..7322
                     /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(1516..1532)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1517..1533)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1535..1782
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1551..1569)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(1804..1822)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1843..1859)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1843..1859)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1856..1878)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1867..1883
                     /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(1891..1897,1898..1915,1916..1921))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1936..1957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2074..2091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2245..2833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2325..2344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3004..3795,3796..3864))
                     /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     3627..3646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3865..3969)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4006..4509
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4550..4568
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4606..4628)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4728..4747
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4771..4986
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4987..5790
                     /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(5921..6376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6058..6079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6270..6289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6502..6524
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6516..6533
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6517..6533
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6540..6559
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6540..6558
                     /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 ttcttcaaga agacctggat catctaagag
     1501 gccttaaaag cttatcgata ccgtcgacct cgagtcatgt aattagttat gtcacgctta
     1561 cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     1621 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     1681 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     1741 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta ccagcttttg
     1801 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     1861 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1921 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1981 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2041 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2101 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3961 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa
     4021 aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa
     4081 agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg
     4141 ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt
     4201 tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc
     4261 gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag
     4321 tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact
     4381 cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca
     4441 cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
     4501 caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4561 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4621 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4681 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4741 gattgtactg agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt
     4801 ttttgatttc ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     4861 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     4921 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga
     4981 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     5041 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     5101 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     5161 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     5221 gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt
     5281 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     5341 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     5401 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     5461 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     5521 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     5581 attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga
     5641 accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt
     5701 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     5761 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     5821 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga
     5881 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     5941 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     6001 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     6061 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     6121 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     6181 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     6241 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6301 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6361 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     6421 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     6481 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     6541 aatacgactc actatagggc gaattggagc tcgccgggat cgaagaaatg atggtaaatg
     6601 aaataggaaa tcaaggagca tgaaggcaaa agacaaatat aagggtcgaa cgaaaaataa
     6661 agtgaaaagt gttgatatga tgtatttggc tttgcggcgc cgaaaaaacg agtttacgca
     6721 attgcacaat catgctgact ctgtggcgga cccgcgctct tgccggcccg gcgataacgc
     6781 tgggcgtgag gctgtgcccg gcggagtttt ttgcgcctgc attttccaag gtttaccctg
     6841 cgctaagggg cgagattgga gaagcaataa gaatgccggt tggggttgcg atgatgacga
     6901 ccacgacaac tggtgtcatt atttaagttg ccgaaagaac ctgagtgcat ttgcaacatg
     6961 agtatactag aagaatgagc caagacttgc gagacgcgag tttgccggtg gtgcgaacaa
     7021 tagagcgacc atgaccttga aggtgagacg cgcataaccg ctagagtact ttgaagagga
     7081 aacagcaata gggttgctac cagtataaat agacaggtac atacaacact ggaaatggtt
     7141 gtctgtttga gtacgctttc aattcatttg ggtgtgcact ttattatgtt acaatatgga
     7201 agggaacttt acacttctcc tatgcacata tattaattaa agtccaatgc tagtagagaa
     7261 ggggggtaac acccctccgc gctcttttcc gatttttttc taaaccgtgg aatatttcgg
     7321 at
//
