LOCUS       sequence-263910-        6795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cherry-PML-IV fusion protein with a mutation in its SIM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6795)
  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 6795)
  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-PML-IV-mSIM-C3
FEATURES             Location/Qualifiers
     source          1..6795
                     /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    3219..3340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3256..3275)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3310..3329
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3347..3802)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3484..3505)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3696..3715
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3829..3933
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3935..4292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3935..4126
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3958..3978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4143..4278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4327..5121
                     /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             4327..5121
                     /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(4381..4400)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4991..5010
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5309..5328)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5353..5400
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5729..6317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6218..6237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6492..6795
                     /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 acaagtactc agatctcgac cccatggagc ctgcacccgc ccgatctccg aggccccagc
     1021 aggaccccgc ccggccccag gagcccacca tgcctccccc cgagaccccc tctgaaggcc
     1081 gccagcccag ccccagcccc agccctacag agcgagcccc cgcttcggag gaggagttcc
     1141 agtttctgcg ctgccagcaa tgccaggcgg aagccaagtg cccgaagctg ctgccttgtc
     1201 tgcacacgct gtgctcagga tgcctggagg cgtcgggcat gcagtgcccc atctgccagg
     1261 cgccctggcc cctaggtgca gacacacccg ccctggataa cgtctttttc gagagtctgc
     1321 agcggcgcct gtcggtgtac cggcagattg tggatgcgca ggctgtgtgc acccgctgca
     1381 aagagtcggc cgacttctgg tgctttgagt gcgagcagct cctctgcgcc aagtgcttcg
     1441 aggcacacca gtggttcctc aagcacgagg cccggcccct agcagagctg cgcaaccagt
     1501 cggtgcgtga gttcctggac ggcacccgca agaccaacaa catcttctgc tccaacccca
     1561 accaccgcac ccctacgctg accagcatct actgccgagg atgttccaag ccgctgtgct
     1621 gctcgtgcgc gctccttgac agcagccaca gtgagctcaa gtgcgacatc agcgcagaga
     1681 tccagcagcg acaggaggag ctggacgcca tgacgcaggc gctgcaggag caggatagtg
     1741 cctttggcgc ggttcacgcg cagatgcacg cggccgtcgg ccagctgggc cgcgcgcgtg
     1801 ccgagaccga ggagctgatc cgcgagcgcg tgcgccaggt ggtagctcac gtgcgggctc
     1861 aggagcgcga gctgctggag gctgtggacg cgcggtacca gcgcgactac gaggagatgg
     1921 ccagtcggct gggccgcctg gatgctgtgc tgcagcgcat ccgcacgggc agcgcgctgg
     1981 tgcagaggat gaagtgctac gcctcggacc aggaggtgct ggacatgcac ggtttcctgc
     2041 gccaggcgct ctgccgcctg cgccaggagg agccccagag cctgcaagct gccgtgcgca
     2101 ccgatggctt cgacgagttc aaggtgcgcc tgcaggacct cagctcttgc atcacccagg
     2161 ggaaagatgc tgcagttagc aagaaagcca gcccagaggc tgccagcact cccagggacc
     2221 ctattgacgt tgacctgccc gaggaggcag agagagtgaa ggcccaggtt caggccctgg
     2281 ggctggctga agcccagcct atggctgtgg tacagtcagt gcccggggca caccccgtgc
     2341 cagtgtacgc cttctccatc aaaggccctt cctatggaga ggatgtctcc aatacaacga
     2401 cagcccagaa gaggaagtgc agccagaccc agtgccccag gaaggtcatc aagatggagt
     2461 ctgaggaggg gaaggaggca aggttggctc ggagctcccc ggagcagccc aggcccagca
     2521 cctccaaggc agtctcacca ccccacctgg atggaccgcc tagccccagg agccccgtca
     2581 taggaagtga ggtcttcctg cccaacagca accacgtggc cagtggcgcc ggggaggcag
     2641 aggaacgcgt tggggggggc agcagctcgg aagactcaga tgccgaaaac tcgtcctccc
     2701 gagagctgga tgacagcagc agtgagtcca gtgacctcca gctggaaggc cccagcaccc
     2761 tcagggtcct ggacgagaac cttgctgacc cccaagcaga agacagacct ctggttttct
     2821 ttgacctcaa gattgacaat gaaagtgggt tctcctgggg ctacccccac ccctttctaa
     2881 tttagtctct gagtcccaaa aagaagtgca ggcagagcca tctgccaggc ccaggagagc
     2941 tctgagctct ggccaacaac tgcagccagg ctgggcagag cactccggct cacctgggct
     3001 cctggcgtgt catttgctgg cttgaataaa gatgtccgcc ttaaaaaaaa gcggccctga
     3061 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     3121 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3181 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3241 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3301 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3361 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3421 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3481 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3541 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3601 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3661 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3721 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3781 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3841 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3901 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3961 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4021 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4141 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4201 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4261 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4321 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4381 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4441 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4501 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4561 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4621 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4681 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4741 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4801 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4861 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4921 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4981 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5041 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5101 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5161 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5221 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5281 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5341 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5401 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5461 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5521 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6361 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6421 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     6481 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6541 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6601 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6661 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6721 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     6781 atcgctatta ccatg
//
