LOCUS       sequence-272324-        8307 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 8307)
  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 8307)
  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..8307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1725..1734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1734..1757
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(4305..4879,4880..4882,4883..4891)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4472..4489)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4699..4718
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4892..5290
                     /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(5329..5348)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5394..5449
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5448..5467)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5810..5826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5810..5826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5823..5845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5834..5850
                     /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(5858..5864,5865..5882,5883..5888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5903..5924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5983..6178
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6029..6164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6184..6318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6221..6240)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6275..6294
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6386..6403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6557..7145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6637..6656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7316..8107,8108..8176))
                     /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     7939..7958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8177..8281)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgccacc atggattaca
     1741 aggatgacga cgataaggga tcaggcatgg aaactgaacc agtttccgtg cagaaggtac
     1801 ctgcaccccc tggatctccc tgtcgacaac aggactctgc cctgaccccc acgcccacca
     1861 tgcctccccc agaggaaccc tccgaagact atgaacacag ccaaagccct gcagagcaag
     1921 ccatacagga ggaatttcag tttctgcgct gcccgagctg ccaggcccaa gccaggtgcc
     1981 ccaaactgct gccttgcctg cacacgctgt gctccggatg cctggaggcg cctggcctgc
     2041 agtgccccat ctgcaaggct cctgggcagg ccgatgctaa tggggaggcc ctggataacg
     2101 tgttcttcga gagcctgcag cggcgcctgg cggtgttccg gcagatcgtg gatgctcagg
     2161 ctgcgtgcac ccgctgcaaa ggcttggccg acttctggtg tttcgagtgt gaacagctca
     2221 tttgcagcaa gtgcttcgaa gcacaccagt ggtacctcag gcatgaggcc cggcccctgg
     2281 ccgatctccg cgacaattca gtgagcagct tcctcgacag tacgcgcaag tccaatatct
     2341 tctgctccaa taccaaccac cgcaaccctg cgctgactga catctactgc cgaggctgcg
     2401 ccaagcctct gtgttgcaca tgcgcgctcc tggaccgcaa ccacagccat ctccattgcg
     2461 atattggtga ggagattcag cagtggcatg aggaactagg caccatgaca cagactctgg
     2521 aggagcaggg cagaaccttc gacagtgccc atgcacagat gtgctcagct ataggacagc
     2581 tggaccacgc acgcgcagac attgagaagc agatccgcgc acgcgtgcgc caggtggtag
     2641 actacgtgca ggctcaggag cgcgagctgc tcgaggcggt gaatgaccgc taccagcgcg
     2701 actaccagga aatagctggc cagctgagct gcctggaagc tgtgctgcag cgcatccgca
     2761 ctagtggggc gctggtcaag aggatgaagc tctatgcctc cgaccaggag gtgctggata
     2821 tgcacagctt tctgcgcaag gcactctgta gcttgcgcca ggaggagccc cagaaccaga
     2881 aagtccagct gctcaccaga ggtttcgagg agttcaagct gtgcctgcag gacttcatct
     2941 cctgcatcac ccagaggata aatgcagctg tagccagccc agaggcagcc agcaatcaac
     3001 cagaggcagc cagcactcac ccagtgacaa ccagcacgcc tgaggacctt gtgcaggagg
     3061 cttctcagac agtcggctcc atgaagagga agtgctccca cgaagattgc tccaggaaga
     3121 tcatccggat ggagtccaca gaggagaacg aggacaggtt ggccacaagc tcccccgagc
     3181 agtcctggcc cagcactttc aaggccacct cccctcccca tctggatggg acttccaacc
     3241 ctgagagcac cgtccctgaa aaaaagatcc tcctgcccaa caacaatcat gttaccagtg
     3301 acacagggga aacagaggag cgagttgtgg tgatcagcag ctcagaggat tcggacaccg
     3361 agaatctgtc ctcccacgag ttggacgata gcagcagtga gtccagcagc ctgcagctgg
     3421 agggccccaa ttccctcaag gccttggatg agagcctcgc cgaaccccac ttagaagaca
     3481 ggaccctggt tttctttgac ctcaagattg acaatgaaac ccagaaaatt agccagctgg
     3541 ccgcggtgaa ccgggaaagc aagttccgtg tgctcatcca gcccgaggcc ttcagtgtct
     3601 actccaaagc tgtctccctg gaggcagggc tccggcactt cctcagcttc ctcaccacca
     3661 tgcatcgtcc catcttggcg tgttctaggc tgtgggggcc gggactcccc atcttcttcc
     3721 agaccctgag tgatattaac aagctgtggg aattccagga caccatctca ggtttcttgg
     3781 ccgtgctgcc cctcatccgg gaacgcatac ccggcgctag cagcttcaaa cttgggaacc
     3841 tagccaagac ctacctggcg agaaacatga gcgaacgcag cgctctggct tctgtgctgg
     3901 ccatgaggga cttgtgctgc ctccttgaga tctccccagg actgccgctg gcccagcata
     3961 tctactcctt tagtagcttg cagtgttttg cttccctgca gcccctaatt caggccagcg
     4021 tcctgccaca gtccgaggcc cgcctcttgg ccctccacaa tgtgagcttt gtagagttgc
     4081 tgaatgcata tcgcaccaac aggcaagaag gcttgaagaa gtatgtccac tatctgagcc
     4141 tgcagaccac cccgttgtca tcgtcggctt ccacccaagt tgcccaattc ctgcaggctc
     4201 ttagcaccca catggaagga ctgttggaag gccatgcccc tgctggggca gaaggcaaag
     4261 ctgaaagtaa gggatgcctg gcctagctcg aggcccggga tccgcccctc tccctccccc
     4321 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     4381 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     4441 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     4501 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     4561 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     4621 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     4681 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     4741 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt
     4801 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     4861 tgaaaaacac gatgataata tggccacaac catggccaag cctttgtctc aagaagaatc
     4921 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     4981 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     5041 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     5101 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     5161 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     5221 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     5281 ggagggttaa tgtacaagta atgccctggc tcacaaatac cactgagatc tttttccctc
     5341 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     5401 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     5461 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     5521 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     5581 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     5641 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     5701 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     5761 tctcttatga agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     5821 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5881 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5941 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     6001 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6061 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6121 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     6181 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6241 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6301 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6361 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6421 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6481 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6541 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6601 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6661 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6721 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     6781 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6841 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6901 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6961 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7021 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7081 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7141 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7201 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7261 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7321 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7381 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7441 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7501 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7561 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7621 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7681 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7741 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7801 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7861 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7921 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7981 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8041 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8101 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8161 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8221 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8281 gcacatttcc ccgaaaagtg ccacctg
//
