LOCUS       sequence-226348-        7286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control vector expressing GFP alone without any consensus repeats.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7286)
  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 7286)
  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-X0
FEATURES             Location/Qualifiers
     source          1..7286
                     /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(1480..1496)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1481..1497)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1499..1746
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1515..1533)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(1768..1786)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1807..1823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1807..1823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1820..1842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1831..1847
                     /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(1855..1861,1862..1879,1880..1885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1900..1921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2038..2055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2209..2797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2289..2308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2968..3759,3760..3828))
                     /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     3591..3610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3829..3933)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3970..4473
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4514..4532
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4570..4592)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4692..4711
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4735..4950
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4951..5754
                     /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(5885..6340)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6022..6043)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6234..6253
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6466..6488
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6480..6497
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6481..6497
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6504..6523
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6504..6522
                     /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 tatacaaata aaagcttatc gataccgtcg acctcgagtc
     1501 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa
     1561 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     1621 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     1681 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     1741 atttgcggcc ggtaccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta
     1801 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1861 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1921 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1981 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2941 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3001 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3061 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3121 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3181 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3241 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3301 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3361 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3421 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3481 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3541 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3601 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3661 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3721 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3781 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3841 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3901 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     3961 gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat
     4021 aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag
     4081 atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct
     4141 ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc
     4201 ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct
     4261 tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta
     4321 tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat
     4381 acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga
     4441 tacgaggcgc gtgtaagtta caggcaagcg atccgtctaa gaaaccatta ttatcatgac
     4501 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4561 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4621 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4681 gcttaactat gcggcatcag agcagattgt actgagagtg caccatacca ccttttcaat
     4741 tcatcatttt ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgattcg
     4801 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     4861 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     4921 acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     4981 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     5041 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     5101 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     5161 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa
     5221 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     5281 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     5341 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     5401 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     5461 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     5521 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     5581 gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt
     5641 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     5701 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     5761 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     5821 cagttattac cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5881 caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5941 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6001 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6061 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6121 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6181 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6241 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6301 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc
     6361 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6421 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6481 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagctcgccg
     6541 ggatcgaaga aatgatggta aatgaaatag gaaatcaagg agcatgaagg caaaagacaa
     6601 atataagggt cgaacgaaaa ataaagtgaa aagtgttgat atgatgtatt tggctttgcg
     6661 gcgccgaaaa aacgagttta cgcaattgca caatcatgct gactctgtgg cggacccgcg
     6721 ctcttgccgg cccggcgata acgctgggcg tgaggctgtg cccggcggag ttttttgcgc
     6781 ctgcattttc caaggtttac cctgcgctaa ggggcgagat tggagaagca ataagaatgc
     6841 cggttggggt tgcgatgatg acgaccacga caactggtgt cattatttaa gttgccgaaa
     6901 gaacctgagt gcatttgcaa catgagtata ctagaagaat gagccaagac ttgcgagacg
     6961 cgagtttgcc ggtggtgcga acaatagagc gaccatgacc ttgaaggtga gacgcgcata
     7021 accgctagag tactttgaag aggaaacagc aatagggttg ctaccagtat aaatagacag
     7081 gtacatacaa cactggaaat ggttgtctgt ttgagtacgc tttcaattca tttgggtgtg
     7141 cactttatta tgttacaata tggaagggaa ctttacactt ctcctatgca catatattaa
     7201 ttaaagtcca atgctagtag agaagggggg taacacccct ccgcgctctt ttccgatttt
     7261 tttctaaacc gtggaatatt tcggat
//
