LOCUS       sequence-425324-        5062 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses reversibly switchable fluorescent protein ffDronpa 
            targeted to the nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5062)
  AUTHORS   Valenta H, Bierbuesse F, Vitale R, Ruckebusch C, Vandenberg W, 
            Dedecker P
  TITLE     Per-pixel unmixing of spectrally overlapping fluorophores using 
            intra-exposure excitation modulation
  JOURNAL   Talanta. 2024 Mar
  PUBMED    38048682
REFERENCE   2  (bases 1 to 5062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Talanta. 
            2024 Mar"
COMMENT     Sequence Label: pH2B-6AA-ffDronpa
FEATURES             Location/Qualifiers
     source          1..5062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        46..349
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        350..553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     504..524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             642..1019
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1038..1712
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     polyA_signal    1835..1956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1872..1891)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1926..1945
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1963..2418)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2100..2121)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2312..2331
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2445..2549
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2551..2908
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2551..2742
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2574..2594)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2759..2894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2943..3737
                     /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             2943..3737
                     /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(2997..3016)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3607..3626
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3925..3944)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3969..4016
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4345..4933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4834..4853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
       61 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      121 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      181 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      241 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      301 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      361 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      421 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      481 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      541 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      601 gagctcaagc ttcgaattct gcagtcgacg gtaccgccac catgccagag ccagcgaagt
      661 ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac taaggcgcag aagaaaggcg
      721 gcaagaagcg caagcgcagc cgcaaggaga gctattccat ctatgtgtac aaggttctga
      781 agcaggtcca ccctgacacc ggcatttcgt ccaaggccat gggcatcatg aattcgtttg
      841 tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat tacaacaagc
      901 gctcgaccat cacctccagg gagatccaga cggccgtgcg cctgctgctg cctggggagt
      961 tggccaagca cgccgtgtcc gagggtacta aggccatcac caagtacacc agcgctaagg
     1021 atccaccggt cgccaccatg agtgtgatta aaccagacat gaagatcaag ctgcgtatgg
     1081 aaggcgctgt aaatggacac ccgttcgcga ttgaaggagt tggccttggg aagcctttcg
     1141 agggaaaaca gagtatggac cttaaagtca aagaaggcgg acctctgcct ttcgcctatg
     1201 acatcttgac aactgtgttc tgttacggca acagggtatt cgccaaatac ccagaaaata
     1261 tagtagacta tttcaagcag tcgtttcctg agggctactc ttgggaacga agcatgaatt
     1321 acgaagacgg gggcatttgt aacgcgacaa acgacataac cctggatggt gactgttata
     1381 tctatgaaat tcgatttgat ggtgtgaact ttcctgccaa tggtccagtt atgcagaaga
     1441 ggactgtgaa atgggagcca tccactgaga aattgtatgt gcgtgatgga gtgctgaagg
     1501 gtgatgttaa catggctctg tcgcttgaag gaggtggcca ttaccgatgt gacttcaaaa
     1561 ctacttataa agctaagaag gttgtccagt tgccagacta tcactttgtg gaccaccaca
     1621 ttgagattaa aagccacgac aaagattaca gtaatgttaa tctgcatgag catgccgaag
     1681 cgcattctga gctgccgagg caggccaagt aaagcggccg cgactctaga tcataatcag
     1741 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1801 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1861 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1921 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     1981 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2041 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2101 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2161 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2221 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2281 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2341 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2401 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2461 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2521 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2581 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2641 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2701 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2761 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2821 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2881 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     2941 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3001 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3061 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3121 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3181 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3241 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3301 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3361 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3421 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3481 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3541 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3601 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3661 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3721 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3781 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3841 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3901 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3961 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4021 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4081 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4141 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4201 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4261 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4321 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4381 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4441 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4501 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4561 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4621 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4681 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4741 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4801 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4861 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4921 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4981 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5041 catgcattag ttattaatag ta
//
