LOCUS       sequence-296009-        8313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse PML KR mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8313)
  AUTHORS   Kurihara M, Kato K, Sanbo C, Shigenobu S, Ohkawa Y, Fuchigami T, 
            Miyanari Y
  TITLE     Genomic Profiling by ALaP-Seq Reveals Transcriptional Regulation by 
            PML Bodies through DNMT3A Exclusion.
  JOURNAL   Mol Cell. 2020 May 7;78(3):493-505.e8. doi: 
            10.1016/j.molcel.2020.04.004. Epub 2020 Apr 29.
  PUBMED    32353257
REFERENCE   2  (bases 1 to 8313)
  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: 
            "10.1016/j.molcel.2020.04.004"; journalName: "Mol Cell"; date: 
            "2020-05-7- 7"; volume: "78"; issue: "3"; pages: "493-505.e8"
COMMENT     Sequence Label: pCFL-mPML-3KRmut-iB
FEATURES             Location/Qualifiers
     source          1..8313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          164..1180
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1104..1126
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1188..1207
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1235..1244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1244..1267
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(3815..4389,4390..4392,4393..4401)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3982..3999)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4209..4228
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4402..4800
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(4839..4858)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4904..4959
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4958..4977)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5320..5336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5320..5336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5333..5355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5344..5360
                     /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)."
     promoter        complement(join(5368..5374,5375..5392,5393..5398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5413..5434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5493..5688
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5539..5674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5694..5828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5731..5750)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5785..5804
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5896..5913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6067..6655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6147..6166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6826..7617,7618..7686))
                     /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     7449..7468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7687..7791)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7822..8201
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8203..8313,1..163)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
       61 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      121 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      181 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      241 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      301 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      361 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
      421 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
      481 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
      541 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
      601 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
      661 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
      721 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
      781 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
      841 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
      901 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
      961 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1021 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1081 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1141 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1201 tattgtgctg tctcatcatt ttggcaaaga attcgccacc atggattaca aggatgacga
     1261 cgataaggga tcaggcatgg aaactgaacc agtttccgtg cagaaggtac ctgcaccccc
     1321 tggatctccc tgtcgacaac aggactctgc cctgaccccc acgcccacca tgcctccccc
     1381 agaggaaccc tccgaagact atgaacacag ccaaagccct gcagagcaag ccatacagga
     1441 ggaatttcag tttctgcgct gcccgagctg ccaggcccaa gccaggtgcc ccaaactgct
     1501 gccttgcctg cacacgctgt gctccggatg cctggaggcg cctggcctgc agtgccccat
     1561 ctgcaaggct cctgggcagg ccgatgctaa tggggaggcc ctggataacg tgttcttcga
     1621 gagcctgcag cggcgcctgg cggtgttccg gcagatcgtg gatgctcagg ctgcgtgcac
     1681 ccgctgcaaa ggcttggccg acttctggtg tttcgagtgt gaacagctca tttgcagcaa
     1741 gtgcttcgaa gcacaccagt ggtacctcag gcatgaggcc cggcccctgg ccgatctccg
     1801 cgacaattca gtgagcagct tcctcgacag tacgcgcaag tccaatatct tctgctccaa
     1861 taccaaccac cgcaaccctg cgctgactga catctactgc cgaggctgcg ccaagcctct
     1921 gtgttgcaca tgcgcgctcc tggaccgcaa ccacagccat ctccattgcg atattggtga
     1981 ggagattcag cagtggcatg aggaactagg caccatgaca cagactctgg aggagcaggg
     2041 cagaaccttc gacagtgccc atgcacagat gtgctcagct ataggacagc tggaccacgc
     2101 acgcgcagac attgagaagc agatccgcgc acgcgtgcgc caggtggtag actacgtgca
     2161 ggctcaggag cgcgagctgc tcgaggcggt gaatgaccgc taccagcgcg actaccagga
     2221 aatagctggc cagctgagct gcctggaagc tgtgctgcag cgcatccgca ctagtggggc
     2281 gctggtcaag aggatgaagc tctatgcctc cgaccaggag gtgctggata tgcacagctt
     2341 tctgcgcaag gcactctgta gcttgcgcca ggaggagccc cagaaccaga aagtccagct
     2401 gctcaccaga ggtttcgagg agttcaagct gtgcctgcag gacttcatct cctgcatcac
     2461 ccagaggata aatgcagctg tagccagccc agaggcagcc agcaatcaac cagaggcagc
     2521 cagcactcac ccagtgacaa ccagcacgcc tgaggacctt gtgcaggagg cttctcagac
     2581 agtcggctcc atgaagagga agtgctccca cgaagattgc tccaggaaga tcatccggat
     2641 ggagtccaca gaggagaacg aggacaggtt ggccacaagc tcccccgagc agtcctggcc
     2701 cagcactttc aaggccacct cccctcccca tctggatggg acttccaacc ctgagagcac
     2761 cgtccctgaa aaaaagatcc tcctgcccaa caacaatcat gttaccagtg acacagggga
     2821 aacagaggag cgagttgtgg tgatcagcag ctcagaggat tcggacaccg agaatctgtc
     2881 ctcccacgag ttggacgata gcagcagtga gtccagcagc ctgcagctgg agggccccaa
     2941 ttccctcaag gccttggatg agagcctcgc cgaaccccac ttagaagaca ggaccctggt
     3001 tttctttgac ctcaagattg acaatgaaac ccagaaaatt agccagctgg ccgcggtgaa
     3061 ccgggaaagc aagttccgtg tgctcatcca gcccgaggcc ttcagtgtct actccaaagc
     3121 tgtctccctg gaggcagggc tccggcactt cctcagcttc ctcaccacca tgcatcgtcc
     3181 catcttggcg tgttctaggc tgtgggggcc gggactcccc atcttcttcc agaccctgag
     3241 tgatattaac aagctgtggg aattccagga caccatctca ggtttcttgg ccgtgctgcc
     3301 cctcatccgg gaacgcatac ccggcgctag cagcttcaaa cttgggaacc tagccaagac
     3361 ctacctggcg agaaacatga gcgaacgcag cgctctggct tctgtgctgg ccatgaggga
     3421 cttgtgctgc ctccttgaga tctccccagg actgccgctg gcccagcata tctactcctt
     3481 tagtagcttg cagtgttttg cttccctgca gcccctaatt caggccagcg tcctgccaca
     3541 gtccgaggcc cgcctcttgg ccctccacaa tgtgagcttt gtagagttgc tgaatgcata
     3601 tcgcaccaac aggcaagaag gcttgaagaa gtatgtccac tatctgagcc tgcagaccac
     3661 cccgttgtca tcgtcggctt ccacccaagt tgcccaattc ctgcaggctc ttagcaccca
     3721 catggaagga ctgttggaag gccatgcccc tgctggggca gaaggcaaag ctgaaagtaa
     3781 gggatgcctg gcctagctcg aggcccggga tccgcccctc tccctccccc ccccctaacg
     3841 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3901 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3961 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     4021 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     4081 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     4141 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     4201 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     4261 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     4321 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     4381 gatgataata tggccacaac catggccaag cctttgtctc aagaagaatc caccctcatt
     4441 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     4501 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     4561 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     4621 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     4681 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     4741 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggttaa
     4801 tgtacaagta atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4861 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4921 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4981 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     5041 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     5101 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     5161 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     5221 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     5281 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5341 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5401 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5461 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     5521 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5581 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5641 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     5701 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5761 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5821 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6961 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7141 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7201 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7261 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7321 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7381 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7441 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7501 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7561 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7621 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7681 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7741 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7801 ccgaaaagtg ccacctgggt cgacattgat tattgactag ttattaatag taatcaatta
     7861 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7921 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7981 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     8041 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     8101 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     8161 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
     8221 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
     8281 tttaattatt ttgtgcagcg atgggggcgg ggg
//
