LOCUS       sequence-472158-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BRD9 Bromodomain as tandem double domain Y57A mutant fused
            to C-terminal part of IFP2.0 as negative control for plasmid 233734.
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_Y57A-IFP2.0C
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     189..208
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(297..654)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      311..446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        463..654
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     611..631
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(656..760)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      787..1242
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(874..893)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1084..1105
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1249..1370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1260..1279)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1314..1333
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    1494..1559
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(2556..2576)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2996..3199)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3025..3045)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3200..3503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3678..4266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3758..3777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4595..4642
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4667..4686
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(4874..5406,1..262))
                     /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             complement(join(4874..5406,1..262))
                     /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(4985..5004)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
       61 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
      121 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
      181 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
      241 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
      301 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
      361 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
      421 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
      481 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      541 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      601 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
      661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      781 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      841 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      901 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      961 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1021 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1081 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1141 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1201 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg
     1261 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1321 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1381 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1441 aaaacctcta caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatccc
     1501 gggcccgcgg taccgtcgac tgcagaattc gaagcttgag ctcgagatct gagtccggat
     1561 taggcttctt tcctctgcac ctgcagggac aggagcctcc ccagatactc cagggtggtc
     1621 cgcagatcgg gtggcagcac gtagggtgtc tgatggtggc agacaatcag tccccacagc
     1681 tggccgccga ccacgacgct gacgctcagg gaggagccca ctcccatatt cctcaggtac
     1741 tgcatgtgca ttggggatgt agctctgagg acagctccgc ccagaggtgt aggagcattt
     1801 gtctggggat tcagcacggg gtcgaggggc actgctgcag ccctggtgtc tgcggtcagc
     1861 cggagcaggt gtcttgtgta gagggcccta gcttgtgcag gggtggtgga tgcgggaaac
     1921 ctatgcccga ggtaagcctg catgccctcg cgccttgctt cggcaataac ctctcctgtt
     1981 gcgtctggtg caaacttgta cagcatcacc cggtcgaaac ctgacagttc ccggactgtt
     2041 tgggttgcga cttcagccag tgcccggaga ttaggagcac tctccagagc gaacattgca
     2101 tttctcagag cgtggggtcc aatagagtcc gtgccctcct ggccttccga cgtggatgaa
     2161 cccgaggttt gtgacgatcc agtcgacatg ctgcgcttca aagctaacag ccgctctttg
     2221 ctcatcatct taaagcctgc gtgaaggatc ttcttcgcca acttgtagta cacggtatct
     2281 ggcctattgt atgtcattgc attatcacac atcagcttga aatctgcctt aaattccgta
     2341 actgacttgt attcattagc tacaattttg tctttcatgg tgccaaaatc catgggatgt
     2401 tttattatca ttgaatatcc aggagcaatt gcatccgtga caggaaaagc aaaaaatcca
     2461 tggggatctt ttctctgaag ctggcggagg aagtgttcca ggagttgctg aataggtgtg
     2521 ctctcatttt cggctgacaa tttaagcatg gtggccacct tgcgtttttt cttgggcgat
     2581 ccagtcgaca tgctgcgctt caaagctaac agccgctctt tgctcatcat cttaaagcct
     2641 gcgtgaagga tcttcttcgc caacttgtag tacacggtat ctggcctatt gtatgtcatt
     2701 gcattatcac acatcagctt gaaatctgcc ttaaattccg taactgactt gtattcatta
     2761 gctacaattt tgtctttcat ggtgccaaaa tccatgggat gttttattat cattgaatat
     2821 ccaggagcaa ttgcatccgt gacaggaaaa gcaaaaaatc catggggatc ttttctctga
     2881 agctggcgga ggaagtgttc caggagttgc tgaataggtg tgctctcatt ttcggctgac
     2941 aatttaagca tggtggcgac cggtagcgct agcggatctg acggttcact aaaccagctc
     3001 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     3061 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     3121 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     3181 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     3241 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     3301 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     3361 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     3421 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     3481 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     3541 taattgatta ctattaataa ctaatgcatg gcggtaatac ggttatccac agaatcaggg
     3601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg
     4441 cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt
     4501 ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca
     4561 gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt
     4621 ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt
     4681 gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg
     4741 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     4801 gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac
     4861 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     4921 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     4981 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     5041 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     5101 cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca
     5161 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     5221 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     5281 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     5341 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     5401 cccttc
//
