LOCUS       sequence-472157-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BRD9 Bromodomain as tandem double domain fused to 
            C-terminal part of IFP2.0 for histone acetylation detection in BiAD 
            sensors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  AUTHORS   Kohler AR, Gutekunst N, Harsch A, Bashtrykov P, Jeltsch A
  TITLE     Development of a BiAD Sensor for Locus-Specific Detection of 
            Cellular Histone Acetylation Dynamics by Fluorescence Microscopy.
  JOURNAL   Genes (Basel). 2025 Apr 10;16(4):444. doi: 10.3390/genes16040444.
  PUBMED    40282404
REFERENCE   2  (bases 1 to 5406)
  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: "Genes 
            (Basel)."; date: "2025-04-10"; volume: "16(4):444. doi"; pages: " 
            10.3390/genes16040444"
COMMENT     Sequence Label: pAK208_2xBRD9BD-IFP2.0C
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             988..1008
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2005..2070
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2194..2315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2231..2250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2285..2304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2322..2777)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2459..2480)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2671..2690
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2804..2908
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2910..3267
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2910..3101
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2933..2953)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3118..3253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3302..4096
                     /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             3302..4096
                     /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(3356..3375)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3966..3985
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4284..4303)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4328..4375
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4704..5292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5193..5212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatgcttaa attgtcagcc gaaaatgaga gcacacctat tcagcaactc
      661 ctggaacact tcctccgcca gcttcagaga aaagatcccc atggattttt tgcttttcct
      721 gtcacggatg caattgctcc tggagcttca atgataataa aacatcccat ggattttggt
      781 accatgaaag acaaaattgt agctaatgaa tacaagtcag ttacggaatt taaggcagat
      841 ttcaagctga tgtgtgataa tgcaatgaca tacaataggc cagataccgt gtactacaag
      901 ttggcgaaga agatccttca cgcaggcttt aagatgatga gcaaagagcg gctgttagct
      961 ttgaagcgca gcatgtcgac tggatcgccc aagaaaaaac gcaaggtggc caccatgctt
     1021 aaattgtcag ccgaaaatga gagcacacct attcagcaac tcctggaaca cttcctccgc
     1081 cagcttcaga gaaaagatcc ccatggattt tttgcttttc ctgtcacgga tgcaattgct
     1141 cctggagctt caatgataat aaaacatccc atggattttg gcaccatgaa agacaaaatt
     1201 gtagctaatg aatacaagtc agttacggaa tttaaggcag atttcaagct gatgtgtgat
     1261 aatgcaatga catacaatag gccagatacc gtgtactaca agttggcgaa gaagatcctt
     1321 cacgcaggct ttaagatgat gagcaaagag cggctgttag ctttgaagcg cagcatgtcg
     1381 actggatcgt cacaaacctc gggttcatcc acgtcggaag gccaggaggg cacggactct
     1441 attggacccc acgctctgag aaatgcaatg ttcgctctgg agagtgctcc taatctccgg
     1501 gcactggctg aagtcgcaac ccaaacagtc cgggaactgt caggtttcga ccgggtgatg
     1561 ctgtacaagt ttgcaccaga cgcaacagga gaggttattg ccgaagcaag gcgcgagggc
     1621 atgcaggctt acctcgggca taggtttccc gcatccacca cccctgcaca agctagggcc
     1681 ctctacacaa gacacctgct ccggctgacc gcagacacca gggctgcagc agtgcccctc
     1741 gaccccgtgc tgaatcccca gacaaatgct cctacacctc tgggcggagc tgtcctcaga
     1801 gctacatccc caatgcacat gcagtacctg aggaatatgg gagtgggctc ctccctgagc
     1861 gtcagcgtcg tggtcggcgg ccagctgtgg ggactgattg tctgccacca tcagacaccc
     1921 tacgtgctgc cacccgatct gcggaccacc ctggagtatc tggggaggct cctgtccctg
     1981 caggtgcaga ggaaagaagc ctaatccgga ctcagatctc gagctcaagc ttcgaattct
     2041 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     2101 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2161 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2221 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2281 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     2341 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2401 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2461 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2521 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2581 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2641 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2701 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2761 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2821 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2881 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2941 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3001 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3121 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3181 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3241 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3301 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3361 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3421 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3481 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3541 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3601 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3661 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3721 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3781 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3841 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3901 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3961 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4021 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4081 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4141 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4201 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4261 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4321 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4381 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4441 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4501 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4561 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4621 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4681 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4741 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4801 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4861 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4921 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4981 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5041 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5101 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5161 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5221 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5281 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5341 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5401 atgcat
//
