LOCUS       sequence-82970-e        8611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL driven LexA-tagged SET1-deltaRRM (RRM domain deleted).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  AUTHORS   Rizzardi LF, Dorn ES, Strahl BD, Cook JG
  TITLE     DNA replication origin function is promoted by H3K4 di-methylation 
            in Saccharomyces cerevisiae.
  JOURNAL   Genetics. 2012 Oct;192(2):371-84. doi: 10.1534/genetics.112.142349. 
            Epub 2012 Jul 30.
  PUBMED    22851644
REFERENCE   2  (bases 1 to 8611)
  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: 
            "10.1534/genetics.112.142349"; journalName: "Genetics"; date: 
            "2012-10"; volume: "192"; issue: "2"; pages: "371-84"
COMMENT     Sequence Label: pGLx2-set1deltaRRM
FEATURES             Location/Qualifiers
     source          1..8611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2190..2207)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2190..2206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2199..2221)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2479..3282)
                     /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"
     promoter        complement(3283..3493)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      3729..4893
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     complement(4910..4929)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5029..5051
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5089..5107)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5175..5279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5280..5348,5349..6140)
                     /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     complement(5498..5517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6311..6899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6800..6819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7053..7070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(7223..7228,7229..7246,7247..7253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7261..7277
                     /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)."
     primer_bind     7266..7288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7285..7301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7285..7301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7525..7966
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    7525..7642
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7937..7960
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(7990..8241,8242..8595)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     8533..8554
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
ORIGIN
        1 gggacaagct acagaagtta caagagaacc tgaagaaggc taaagagaaa gaagcagaaa
       61 acgaaaaagc aaaggaatta cagggcaaag atattacctt gcccaaggaa cctaaggtag
      121 acacattatc tcattcgtcc ggaagtgaaa aaagaattcc atatgatctc ttgggggtag
      181 ttaataacag acctgtttta catgtctcca aaatatttgt tgccaaacat aggttctgcg
      241 ttgaggactt taaatacaag ttaaggggat acagatgtgc gaaatttatt gatcatccaa
      301 ctggtatcta tattattttt aatgacattg cccatgcgca aacatgttcg aatgcagagt
      361 caggaaattt aacaataatg tctcggagca gaagaattcc tattctaata aagtttcatc
      421 tcattctccc taggttccaa aacagaacta gattcaataa atctagctca tcttcaaatt
      481 ctacaaatgt acctataaaa tacgagtcca aagaggagtt cattgaagct acagcaaaac
      541 aaatattaaa agatttggaa aagactttac atgttgatat taagaagaga ttgattggtc
      601 ctacggtatt tgatgctttg gaccatgcaa attttcctga attgttagct aaaagagaac
      661 taaaggagaa agagaagaga caacagattg catctaaaat tgctgaagat gaattgaaac
      721 gtaaagaaga agccaaaaga gattttgatt tgtttggttt atatggtggc tatgcaaaat
      781 ctaataaaag aaatttaaaa aggcataatt cactcgcgtt ggatcatact tctttaaaga
      841 ggaaaaagct atccaatggt atcaaaccaa tggcacattt actgaacgaa gaaaccgatt
      901 ccaaagaaac taccccattg aacgatgaag ggatcactcg cgtatcaaaa gaacatgatg
      961 aagaagacga aaatatgaca tcttcatctt ctgaagaaga ggaagaagaa gctccagata
     1021 agaaattcaa gagtgagtct gagccaacca cccccgaatc tgatcacctt catggtatta
     1081 agccgttagt acccgatcaa aatgggtcgt ctgacgtact ggatgcttct tcgatgtata
     1141 aacctactgc taccgaaatt cccgaacctg tatatccacc tgaggaatat gacttgaaat
     1201 atagtcagac tttatcttct atggatttgc agaatgctat caaagatgag gaagatatgc
     1261 taattttaaa gcagttattg agcacatata ctcctaccgt cacaccagaa acaagcgcag
     1321 ctctggaata taaaatttgg caatctcgcc gaaaagttct tgaagaagag aaggcttccg
     1381 attggcaaat agagcttaat ggaactttat ttgatagtga actacaacca ggtagctctt
     1441 ttaaagctga agggttcagg aaaattgcgg ataaattaaa aattaattac ctaccccatc
     1501 gtcgcagagt tcaccaacct ttaaatacgg tgaatattca caatgaaagg aatgagtaca
     1561 cacctgaact ttgtcaaaga gaagaatcct cgaataaaga accttcagac tcagttcctc
     1621 aagaagtttc atcctctaga gataataggg catcaaatag aagatttcag caggacatag
     1681 aggcacagaa agccgcaatt ggtacggaat ctgagctgct atcactaaat caattaaata
     1741 aaagaaaaaa gccagttatg ttcgctcgtt cagcaattca caactggggt ttatatgctc
     1801 tagactctat cgcagcaaag gaaatgatta tcgagtacgt tggtgaaagg atcaggcaac
     1861 ctgtagcaga aatgagagag aaaagatatc tgaaaaatgg gattggatcc agttaccttt
     1921 ttagggttga tgaaaacacg gttattgatg ccaccaagaa aggtggtata gcccgtttca
     1981 ttaatcattg ttgtgatcca aattgtacgg caaagattat aaaggttggc gggagaagga
     2041 gaattgttat ctatgcactg cgtgatatcg cggcaagcga agagttgaca tatgattaca
     2101 aatttgagag agaaaaggat gacgaggaaa gacttccttg tttatgtgga gcacctaatt
     2161 gtaaaggttt cttgaactga aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     2221 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccctt cgccagctgg
     2281 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2341 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2401 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     2461 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     2521 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     2581 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     2641 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     2701 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     2761 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     2821 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     2881 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     2941 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     3001 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     3061 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     3121 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     3181 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     3241 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcgg
     3301 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     3361 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     3421 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa
     3481 atgatgaatt gaaaagctct tgttacccat cattgaattt tgaacatccg aacctgggag
     3541 ttttccctga aacagatagt atatttgaac ctgtataata atatatagtc tagcgcttta
     3601 cggaagacaa tgtatgtatt tcggttcctg gagaaactat tgcatctatt gcataggtaa
     3661 tcttgcacgt cgcatccccg gttcattttc tgcgtttcca tcttgcactt caatagcata
     3721 tctttgttaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc
     3781 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag
     3841 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag
     3901 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     3961 gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc
     4021 ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct
     4081 ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc
     4141 tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact
     4201 gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct
     4261 ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca
     4321 ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa
     4381 tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt
     4441 tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa
     4501 gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag
     4561 caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg
     4621 ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc
     4681 aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcg gaataggaac
     4741 ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag
     4801 ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag
     4861 aacggcatag tgcgtgttta tgcttaaatg cgttatggtg cactctcagt acaatctgct
     4921 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4981 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5041 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5101 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5161 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5221 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5461 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5581 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5821 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5881 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6121 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6421 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     6721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7081 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tccagctggc
     7141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttacc
     7201 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7261 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaattcgaat
     7321 tttcaaaaat tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca
     7381 tggcattacc accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa
     7441 agagccccat tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta
     7501 actgctcatt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
     7561 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
     7621 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
     7681 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
     7741 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
     7801 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
     7861 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
     7921 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aacggatcat
     7981 tcggccgcca tgaaagcgtt aacggccagg caacaagagg tgtttgatct catccgtgat
     8041 cacatcagcc agacaggtat gccgccgacg cgtgcggaaa tcgcgcagcg tttggggttc
     8101 cgttccccaa acgcggctga agaacatctg aaggcgctgg cacgcaaagg cgttattgaa
     8161 attgtttccg gcgcatcacg cgggattcgt ctgttgcagg aagaggaaga agggttgccg
     8221 ctggtaggtc gtgtggctgc cggtgaacca cttctggcgc aacagcatat tgaaggtcat
     8281 tatcaggtcg atccttcctt attcaagccg aatgctgatt tcctgctgcg cgtcagcggg
     8341 atgtcgatga aagatatcgg cattatggat ggtgacttgc tggcagtgca taaaactcag
     8401 gatgtacgta acggtcaggt cgttgtcgca cgtattgatg acgaagttac cgttaagcgc
     8461 ctgaaaaaac agggcaataa agtcgaactg ttgccagaaa atagcgagtt taaaccaatt
     8521 gtcgtagatc ttcgtcagca gagcttcacc attgaagggc tggcggttgg ggttattcgc
     8581 aacggcgact ggctgggatc ctcgagcgcc c
//
