LOCUS       sequence-429908-        5538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses double domain of DNMT3A PWWP domain fused to C-terminal 
            part of IFP2.0 for H3K36me2/3 detection in BiAD sensors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5538)
  AUTHORS   Kohler AR, Hausser J, Harsch A, Bernhardt S, Haussermann L, Brenner 
            LM, Lungu C, Olayioye MA, Bashtrykov P, Jeltsch A
  TITLE     Modular dual-color BiAD sensors for locus-specific readout of 
            epigenome modifications in single cells.
  JOURNAL   Cell Rep Methods. 2024 Mar 26:100739. doi: 
            10.1016/j.crmeth.2024.100739.
  PUBMED    38554702
REFERENCE   2  (bases 1 to 5538)
  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: "Cell Rep 
            Methods. 2024 Mar 26:100739. doi: 10.1016/j.crmeth.2024.100739."
COMMENT     Sequence Label: 2x3APWWP-IFP2.0C
FEATURES             Location/Qualifiers
     source          1..5538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1056..1076
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1077..1086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2139..2204
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2328..2449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2365..2384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2419..2438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2456..2911)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2593..2614)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2805..2824
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2938..3042
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3044..3401
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3044..3235
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3067..3087)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3252..3387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3436..4230
                     /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             3436..4230
                     /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(3490..3509)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4100..4119
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4418..4437)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4462..4509
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4838..5426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5327..5346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      601 taccggtcgc caccatggag tacgaggacg gccggggctt tggcattggg gagctggtgt
      661 gggggaaact gcggggcttc tcctggtggc caggccgcat tgtgtcttgg tggatgacgg
      721 gccggagccg agcagctgaa ggcacccgct gggtcatgtg gttcggagac ggcaaattct
      781 cagtggtgtg tgttgagaag ctgatgccgc tgagctcgtt ttgcagtgcg ttccaccagg
      841 ccacgtacaa caagcagccc atgtaccgca aagccatcta cgaggtcctg caggtggcca
      901 gcagccgcgc ggggaagctg ttcccggtgt gccacgacag cgatgagagt gacactgcca
      961 aggccgtgga ggtgcagaac aagcccatga ttgaatgggc cctggggggc ttccagcctt
     1021 ctggccctaa gggcctggaa ccaccaactg gatcgcccaa gaaaaaacgc aaggtggcca
     1081 ccatggagta tgaggacggc cggggctttg gcattgggga gctggtgtgg gggaaactgc
     1141 ggggcttctc ctggtggcca ggccgcattg tgtcttggtg gatgacgggc cggagccgag
     1201 cagctgaagg cacccgctgg gtcatgtggt tcggagacgg caaattctca gtggtgtgtg
     1261 ttgagaagct gatgccgctg agctcgtttt gcagtgcgtt ccaccaggcc acgtacaaca
     1321 agcagcccat gtaccgcaaa gccatctacg aggtcctgca ggtggccagc agccgcgcgg
     1381 ggaagctgtt cccggtgtgc cacgacagcg atgagagtga cactgccaag gccgtggagg
     1441 tgcagaacaa gcccatgatt gaatgggccc tggggggctt ccagccttct ggccctaagg
     1501 gcctggagcc accaactgga tcgtcacaaa cctcgggttc atccacgtcg gagggccagg
     1561 agggcacgga ctctattgga ccccacgctc tgagaaatgc aatgttcgct ctggagagtg
     1621 ctcctaatct ccgggcactg gctgaagtcg caacccaaac agtccgggaa ctgtcaggtt
     1681 tcgaccgggt gatgctgtac aagtttgcac cagacgcaac aggagaggtt attgccgaag
     1741 caaggcgcga gggcatgcag gcttacctcg ggcataggtt tcccgcatcc accacccctg
     1801 cacaagctag ggccctctac acaagacacc tgctccggct gaccgcagac accagggctg
     1861 cagcagtgcc cctcgacccc gtgctgaatc cccagacaaa tgctcctaca cctctgggcg
     1921 gagctgtcct cagagctaca tccccaatgc acatgcagta cctgaggaat atgggagtgg
     1981 gctcctccct gagcgtcagc gtcgtggtcg gcggccagct gtggggactg attgtctgcc
     2041 accatcagac accctacgtg ctgccacccg atctgcggac caccctggag tatctgggga
     2101 ggctcctgtc cctgcaggtg cagaggaaag aagcttaatc cggactcaga tctcgagctc
     2161 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
     2221 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2281 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2341 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2401 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2461 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2521 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2581 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2641 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2701 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2761 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2821 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2881 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2941 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3001 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3061 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3121 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3181 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3241 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3301 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3361 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3421 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3481 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3541 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3601 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3661 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3721 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3781 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3841 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3901 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3961 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4021 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4081 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4141 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4201 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4261 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4321 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4381 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4441 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4501 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4561 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4621 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4681 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4741 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4801 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4861 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4921 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4981 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5041 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5101 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5161 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5221 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5281 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5341 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5401 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5461 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5521 accgtattac cgccatgc
//
