LOCUS       sequence-240359-        5074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H3K79M expression plasmid for norleucine supplementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5074)
  AUTHORS   Worden EJ, Hoffmann NA, Hicks CW, Wolberger C
  TITLE     Mechanism of Cross-talk between H2B Ubiquitination and H3 
            Methylation by Dot1L.
  JOURNAL   Cell. 2019 Feb 8. pii: S0092-8674(19)30151-5. doi: 
            10.1016/j.cell.2019.02.002.
  PUBMED    30765112
REFERENCE   2  (bases 1 to 5074)
  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"; journalName: "Cell. 2019 
            Feb 8. pii: S0092-8674(19)30151-5. doi: 10.1016/j.cell.2019.02.002."
COMMENT     Sequence Label: pEW65: pQE-81L H3(K79M)
FEATURES             Location/Qualifiers
     source          1..5074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..29
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(53..67,68..73,74..90,91..97)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    73..89
                     /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)."
     protein_bind    105..121
                     /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)."
     RBS             140..151
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      574..668
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             712..1371
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(778..797)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1436..1522
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1615..2697)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2659..2678
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2698..2775)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(2859..2881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2967..3107
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3022..3041
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3122..3139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3293..3881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3373..3392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4052..4843,4844..4912))
                     /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     4675..4694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4913..5017)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgaga aatcataaaa
       61 aatttatttg ctttgtgagc ggataacaat tataatagat tcaattgtga gcggataaca
      121 atttcacaca gaattcatta aagaggagaa attaactatg gcccgtacca agcagaccgc
      181 ccgtaaatcc accggaggga aggctccccg caagcagctg gccaccaagg cagccaggaa
      241 gtccgctcct gctaccggcg gagtcaagaa acctcaccgt taccggcccg gcacagtcgc
      301 tctccgcgag atccgccgct accagaaatc caccgagctg ctcatccgca aactgccttt
      361 ccagcgcctg gtccgggaga tcgctcagga cttcatgacc gacctgcgct tccagagctc
      421 ggccgttatg gctctgcagg aggccagcga ggcttatctg gtcgctctct ttgaggacac
      481 caacctgtgc gccatccacg ccaagagggt caccatcatg cccaaggaca tccagctggc
      541 ccgcagaatc cgaggcgaga gggcttaggc ttggactcct gttgatagat ccagtaatga
      601 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
      661 gtgagaatcc aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
      721 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
      781 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
      841 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
      901 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
      961 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1021 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1081 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1141 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1201 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1261 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1321 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     1381 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     1441 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1501 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     1561 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     1621 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1681 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     1741 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     1801 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     1861 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     1921 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     1981 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2041 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2101 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2161 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2221 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2281 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2341 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2401 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2461 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2521 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     2581 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     2641 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     2701 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     2761 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     2821 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2881 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2941 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3001 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     3061 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3121 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3181 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3241 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3301 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3361 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3421 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3481 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3541 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3601 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3661 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3721 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3781 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3961 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4021 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4081 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4141 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4201 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4261 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4321 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4381 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4441 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4501 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4561 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4621 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4681 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4741 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4801 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4861 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5041 ctgacgtcta agaaaccatt attatcatga catt
//
