LOCUS       sequence-264206-        6946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP-DAXX fusion protein with a mutations in both its 5' 
            and 3' SIMs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  AUTHORS   Shastrula PK, Sierra I, Deng Z, Keeney F, Hayden JE, Lieberman PM, 
            Janicki SM
  TITLE     PML is recruited to heterochromatin during S phase and represses 
            DAXX-mediated histone H3.3 chromatin assembly.
  JOURNAL   J Cell Sci. 2019 Mar 26;132(6). pii: jcs.220970. doi: 
            10.1242/jcs.220970.
  PUBMED    30796101
REFERENCE   2  (bases 1 to 6946)
  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: "J Cell 
            Sci."; date: "2019-03-26"; volume: "132(6). pii: jcs.220970. doi"; 
            pages: " 10.1242/jcs.220970"
COMMENT     Sequence Label: YFP-DAXX-mSIM1/2
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3370..3491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3461..3480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3498..3953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3635..3656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3847..3866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4086..4443
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4086..4277
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4109..4129)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4294..4429
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4478..5272
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4478..5272
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4532..4551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5142..5161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5460..5479)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5504..5551
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5880..6468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6369..6388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6643..6946
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ttcggctacg gcctgaagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg gacgagctgt
      961 acaagtccgg actcagatct cgagccatgg ccaccgctaa cagcatcggc gcgggggatg
     1021 atgatgacga agatgaagca gctgctcagc cagggccctc ccacccactc cccaatgcgg
     1081 cctcacctgg ggcagaagcc cctagctcct ctgagcctca tgggaccaga ggaagcagta
     1141 gttcgggcgg caagaaatgc tacaagctgg agaatgagaa gctgttcgaa gagttccttg
     1201 aactttgtaa gatgcagaca gcagaccacc ctgaggtggt cccattcctc tataaccggc
     1261 agcaacgtgc ccactctctg tttttggcct cggcggagtt ctgcaacatc ctctctaggg
     1321 tcctgtctcg ggcccggagc cggccagcca agctctatgt ctacatcaat gagctctgca
     1381 ctgttctcaa ggcccactca gccaaaaaga agctgaactt ggcccctgcc gccaccacct
     1441 ccaatgagcc ctctgggaat aaccctccca cacacctctc cttggacccc acaaatgctg
     1501 aaaacactgc ctctcagtct ccaaggaccc gtggttcccg gcggcagatc cagcgtttgg
     1561 agcagctgct ggcgctctat gtggcagaga tccggcggct gcaggaaaag gagttagatc
     1621 tgagcgaatt ggatgaccca gactccgcat acctgcagga ggcacggttg aagcgtaagc
     1681 tgatccgcct ctttgggcga ctatgtgagc tgaaagactg ctcttcactg accggccgtg
     1741 tcatagagca gcgcatcccc taccgtggca cccgctaccc agaggttaac aggcgcattg
     1801 agcggctcat caacaagcca gggcctgata ccttccctga ctatggggat gtgcttcggg
     1861 ctgtagagaa ggcagctgcc cgacacagcc ttggcctccc ccgacagcag ctccagctca
     1921 tggctcagga tgccttccga gatgtgggca tcaggttaca ggagcgacgt cacctcgatc
     1981 tcatctacaa ctttggctgc cacctcacag atgactatag gccaggcgtt gaccctgcac
     2041 tatcagatcc tgtgttggcc cggcgccttc gggaaaaccg gagtttggcc atgagtcggc
     2101 tggatgaggt catctccaaa tacgcaatgt tgcaagacaa aagtgaggag ggcgagagaa
     2161 aaaagagaag agctcggctc caaggcacct cttcccactc tgcagacacc cccgaagcct
     2221 ccttggattc tggtgagggc cctagtggaa tggcatccca ggggtgccct tctgcctcca
     2281 gagctgagac agatgacgaa gacgatgagg agagtgatga ggaagaggag gaggaggagg
     2341 aagaagaaga ggaggaggcc acagattctg aagaggagga ggatctggaa cagatgcagg
     2401 agggtcagga ggatgatgaa gaggaggacg aagaggaaga agcagcagca ggtaaagatg
     2461 gagacaagag ccccatgtcc tcactacaga tctccaatga aaagaacctg gaacctggca
     2521 aacagatcag cagatcttca ggggagcagc aaaacaaagg acgcatagtg tcaccatcgt
     2581 tactgtcaga agaacccctg gccccctcca gcatagatgc tgaaagcaat ggagaacagc
     2641 ctgaggagct gaccctggag gaagaaagcc ctgtgtctca gctctttgag ctagagattg
     2701 aagctttgcc cctggatacc ccttcctctg tggagacgga catttcctct tccaggaagc
     2761 aatcagagga gcccttcacc actgtcttag agaatggagc aggcatggtc tcttctactt
     2821 ccttcaatgg aggcgtctct cctcacaact ggggagattc tggtcccccc tgcaaaaaat
     2881 ctcggaagga gaagaagcaa acaggatcag ggccattagg aaacagctat gtggaaaggc
     2941 aaaggtcagt gcatgagaag aatgggaaaa agatatgtac cctgcccagc ccaccttccc
     3001 ccttggcttc cttggcccca gttgctgatt cctccacgag ggtggactct cccagccatg
     3061 gcctggtgac cagctccctc tgcatccctt ctccagcccg gctgtcccaa accccccatt
     3121 cacagcctcc tcggcctggt acttgcaaga caagtgtggc cacacaatgc gatccagaag
     3181 agatcggcgc gggctcagac tctgattagg aattctgcag tcgacggtac cgcgggcccg
     3241 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     3301 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3361 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3481 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3541 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3601 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3661 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3721 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3781 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3841 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3901 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4081 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4321 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4381 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4441 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4501 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4561 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4621 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4681 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4741 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4801 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4861 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4921 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4981 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5041 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5101 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5161 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5221 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5281 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5341 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5401 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5461 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5521 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5581 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5641 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5701 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6541 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
     6601 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6661 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6721 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     6781 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     6841 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6901 tcctacttgg cagtacatct acgtattagt catcgctatt accatg
//
