LOCUS       sequence-296022-        8313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse PML Ca 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-2CAMut-iB
FEATURES             Location/Qualifiers
     source          1..8313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          159..1175
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1099..1121
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1183..1202
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1230..1239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1239..1262
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(3810..4384,4385..4387,4388..4396)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3977..3994)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4204..4223
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4397..4795
                     /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(4834..4853)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4899..4954
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4953..4972)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5315..5331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5315..5331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5328..5350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5339..5355
                     /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(5363..5369,5370..5387,5388..5393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5408..5429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5488..5683
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5534..5669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5689..5823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5726..5745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5780..5799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5891..5908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6062..6650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6142..6161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6821..7612,7613..7681))
                     /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     7444..7463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7682..7786)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7817..8196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8198..8313,1..158)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
       61 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      121 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      181 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      241 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      301 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
      361 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
      421 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
      481 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
      541 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
      601 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
      661 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
      721 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
      781 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
      841 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
      901 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
      961 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1021 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     1081 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     1141 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     1201 tgctgtctca tcattttggc aaagaattcg ccaccatgga ttacaaggat gacgacgata
     1261 agggatcagg catggaaact gaaccagttt ccgtgcagaa ggtacctgca ccccctggat
     1321 ctccctgtcg acaacaggac tctgccctga cccccacgcc caccatgcct cccccagagg
     1381 aaccctccga agactatgaa cacagccaaa gccctgcaga gcaagccata caggaggaat
     1441 ttcagtttct gcgcgcaccg agcgcacagg cccaagccaa gtgccccaaa ctgctgcctt
     1501 gcctgcacac gctgtgctcc ggatgcctgg aggcgcctgg cctgcagtgc cccatctgca
     1561 aggctcctgg gcaggccgat gctaatgggg aggccctgga taacgtgttc ttcgagagcc
     1621 tgcagcggcg cctggcggtg ttccggcaga tcgtggatgc tcaggctgcg tgcacccgct
     1681 gcaaaggctt ggccgacttc tggtgtttcg agtgtgaaca gctcatttgc agcaagtgct
     1741 tcgaagcaca ccagtggtac ctcaagcatg aggcccggcc cctggccgat ctccgcgaca
     1801 attcagtgag cagcttcctc gacagtacgc gcaagtccaa tatcttctgc tccaatacca
     1861 accaccgcaa ccctgcgctg actgacatct actgccgagg ctgcgccaag cctctgtgtt
     1921 gcacatgcgc gctcctggac cgcaaccaca gccatctcca ttgcgatatt ggtgaggaga
     1981 ttcagcagtg gcatgaggaa ctaggcacca tgacacagac tctggaggag cagggcagaa
     2041 ccttcgacag tgcccatgca cagatgtgct cagctatagg acagctggac cacgcacgcg
     2101 cagacattga gaagcagatc cgcgcacgcg tgcgccaggt ggtagactac gtgcaggctc
     2161 aggagcgcga gctgctcgag gcggtgaatg accgctacca gcgcgactac caggaaatag
     2221 ctggccagct gagctgcctg gaagctgtgc tgcagcgcat ccgcactagt ggggcgctgg
     2281 tcaagaggat gaagctctat gcctccgacc aggaggtgct ggatatgcac agctttctgc
     2341 gcaaggcact ctgtagcttg cgccaggagg agccccagaa ccagaaagtc cagctgctca
     2401 ccagaggttt cgaggagttc aagctgtgcc tgcaggactt catctcctgc atcacccaga
     2461 ggataaatgc agctgtagcc agcccagagg cagccagcaa tcaaccagag gcagccagca
     2521 ctcacccagt gacaaccagc acgcctgagg accttgtgca ggaggcttct cagacagtcg
     2581 gctccatgaa gaggaagtgc tcccacgaag attgctccag gaagatcatc aagatggagt
     2641 ccacagagga gaacgaggac aggttggcca caagctcccc cgagcagtcc tggcccagca
     2701 ctttcaaggc cacctcccct ccccatctgg atgggacttc caaccctgag agcaccgtcc
     2761 ctgaaaaaaa gatcctcctg cccaacaaca atcatgttac cagtgacaca ggggaaacag
     2821 aggagcgagt tgtggtgatc agcagctcag aggattcgga caccgagaat ctgtcctccc
     2881 acgagttgga cgatagcagc agtgagtcca gcagcctgca gctggagggc cccaattccc
     2941 tcaaggcctt ggatgagagc ctcgccgaac cccacttaga agacaggacc ctggttttct
     3001 ttgacctcaa gattgacaat gaaacccaga aaattagcca gctggccgcg gtgaaccggg
     3061 aaagcaagtt ccgtgtgctc atccagcccg aggccttcag tgtctactcc aaagctgtct
     3121 ccctggaggc agggctccgg cacttcctca gcttcctcac caccatgcat cgtcccatct
     3181 tggcgtgttc taggctgtgg gggccgggac tccccatctt cttccagacc ctgagtgata
     3241 ttaacaagct gtgggaattc caggacacca tctcaggttt cttggccgtg ctgcccctca
     3301 tccgggaacg catacccggc gctagcagct tcaaacttgg gaacctagcc aagacctacc
     3361 tggcgagaaa catgagcgaa cgcagcgctc tggcttctgt gctggccatg agggacttgt
     3421 gctgcctcct tgagatctcc ccaggactgc cgctggccca gcatatctac tcctttagta
     3481 gcttgcagtg ttttgcttcc ctgcagcccc taattcaggc cagcgtcctg ccacagtccg
     3541 aggcccgcct cttggccctc cacaatgtga gctttgtaga gttgctgaat gcatatcgca
     3601 ccaacaggca agaaggcttg aagaagtatg tccactatct gagcctgcag accaccccgt
     3661 tgtcatcgtc ggcttccacc caagttgccc aattcctgca ggctcttagc acccacatgg
     3721 aaggactgtt ggaaggccat gcccctgctg gggcagaagg caaagctgaa agtaagggat
     3781 gcctggccta gctcgaggcc cgggatccgc ccctctccct cccccccccc taacgttact
     3841 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     3901 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     3961 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     4021 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     4081 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     4141 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     4201 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     4261 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
     4321 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     4381 taatatggcc acaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     4441 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     4501 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     4561 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     4621 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     4681 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     4741 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gttaatgtac
     4801 aagtaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     4861 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     4921 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     4981 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     5041 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     5101 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     5161 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     5221 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     5281 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5341 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5401 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5461 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     5521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     5701 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5761 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5821 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5881 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7801 aagtgccacc tgggtcgaca ttgattattg actagttatt aatagtaatc aattacgggg
     7861 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     7921 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     7981 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     8041 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     8101 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     8161 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     8221 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     8281 ttattttgtg cagcgatggg ggcggggggg ggg
//
