LOCUS       sequence-236153-        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"
     misc_feature    84..224
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     139..158
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(239..256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(410..998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(490..509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1169..1960,1961..2029))
                     /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     1792..1811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2030..2134)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2202..2220
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2244..2258,2259..2264,2265..2281,2282..2288)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2264..2280
                     /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    2296..2312
                     /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             2331..2342
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2765..2859
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2903..3562
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2969..2988)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3627..3713
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3806..4888)
                     /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     4850..4869
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4889..4966)
                     /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(5050..5072)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
       61 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
      121 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
      181 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
      241 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      301 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      361 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      421 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      481 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      541 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      841 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      961 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1021 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1081 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1141 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1201 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1261 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1321 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1381 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1441 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1501 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1561 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1621 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1681 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1741 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1801 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1861 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1921 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1981 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2041 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2101 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2161 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2221 cctttcgtct tcacctcgag aaatcataaa aaatttattt gctttgtgag cggataacaa
     2281 ttataataga ttcaattgtg agcggataac aatttcacac agaattcatt aaagaggaga
     2341 aattaactat ggcccgtacc aagcagaccg cccgtaaatc caccggaggg aaggctcccc
     2401 gcaagcagct ggccaccaag gcagccagga agtccgctcc tgctaccggc ggagtcaaga
     2461 aacctcaccg ttaccggccc ggcacagtcg ctctccgcga gatccgccgc taccagaaat
     2521 ccaccgagct gctcatccgc aaactgcctt tccagcgcct ggtccgggag atcgctcagg
     2581 acttcatgac cgacctgcgc ttccagagct cggccgttat ggctctgcag gaggccagcg
     2641 aggcttatct ggtcgctctc tttgaggaca ccaacctgtg cgccatccac gccaagaggg
     2701 tcaccatcat gcccaaggac atccagctgg cccgcagaat ccgaggcgag agggcttagg
     2761 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     2821 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagctagct tggcgagatt
     2881 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     2941 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     3001 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     3061 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     3121 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     3181 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     3241 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     3301 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     3361 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     3421 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     3481 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     3541 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     3601 ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3661 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct
     3721 ctagattacg tgcagtcgat gataagctgt caaacatgag aattgtgcct aatgagtgag
     3781 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3841 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     3901 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     3961 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     4021 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     4081 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     4141 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     4201 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     4261 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     4321 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     4381 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     4441 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     4501 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     4561 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     4621 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     4681 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     4741 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     4801 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     4861 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     4921 gctatcatgc cataccgcga aaggttttgc accattcgat ggtgtcggaa tttcgggcag
     4981 cgttgggtcc tggccacggg tgcgcatgat ctagagctgc ctcgcgcgtt tcggtgatga
     5041 cggtgaaaac ctctgacaca tgcagctccc ggag
//
