LOCUS       sequence-425328-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses reversibly switchable fluorescent protein ffDronpaF 
            targeted to vimentin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  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 6065)
  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: "Talanta. 
            2024 Mar"
COMMENT     Sequence Label: pvimentin-ffDronpaF
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1996..2670
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVI
                     EGGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDVKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     polyA_signal    2793..2914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2830..2849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2884..2903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2921..3376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3270..3289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3403..3507
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3509..3866
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3509..3700
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3532..3552)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3717..3852
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3901..4695
                     /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             3901..4695
                     /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(3955..3974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4565..4584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4883..4902)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4927..4974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5303..5891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5792..5811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcccatgt ccaccaggtc cgtgtcctcg
      601 tcctcctacc gcaggatgtt cggcggcccg ggcaccgcga gccggccgag ctccagccgg
      661 agctacgtga ctacgtccac ccgcacctac agcctgggca gcgcgctgcg ccccagcacc
      721 agccgcagcc tctacgcctc gtccccgggc ggcgtgtatg ccacgcgctc ctctgccgtg
      781 cgcctgcgga gcagcgtgcc cggggtgcgg ctcctgcagg actcggtgga cttctcgctg
      841 gccgacgcca tcaacaccga gttcaagaac acccgcacca acgagaaggt ggagctgcag
      901 gagctgaatg accgcttcgc caactacatc gacaaggtgc gcttcctgga gcagcagaat
      961 aagatcctgc tggccgagct cgagcagctc aagggccaag gcaagtcgcg cctgggggac
     1021 ctctacgagg aggagatgcg ggagctgcgc cggcaggtgg accagctaac caacgacaaa
     1081 gcccgcgtcg aggtggagcg cgacaacctg gccgaggaca tcatgcgcct ccgggagaaa
     1141 ttgcaggagg agatgcttca gagagaggaa gccgaaaaca ccctgcaatc tttcagacag
     1201 gatgttgaca atgcgtctct ggcacgtctt gaccttgaac gcaaagtgga atctttgcaa
     1261 gaagagattg cctttttgaa gaaactccac gaagaggaaa tccaggagct gcaggctcag
     1321 attcaggaac agcatgtcca aatcgatgtg gatgtttcca agcctgacct cacggctgcc
     1381 ctgcgtgacg tacgtcagca atatgaaagt gtggctgcca agaacctgca ggaggcagaa
     1441 gaatggtaca aatccaagtt tgctgacctc tctgaggctg ccaaccggaa caatgacgcc
     1501 ctgcgccagg caaagcagga gtccactgag taccggagac aggtgcagtc cctcacctgt
     1561 gaagtggatg cccttaaagg aaccaatgag tccctggaac gccagatgcg tgaaatggaa
     1621 gagaactttg ccgttgaagc tgctaactac caagacacta ttggccgcct gcaggatgag
     1681 attcagaata tgaaggagga aatggctcgt caccttcgtg aataccaaga cctgctcaat
     1741 gttaagatgg cccttgacat tgagattgcc acctacagga agctgctgga aggcgaggag
     1801 agcaggattt ctctgcctct tccaaacttt tcctccctga acctgaggga aactaatctg
     1861 gattcactcc ctctggttga tacccactca aaaaggacac ttctgattaa gacggttgaa
     1921 actagagatg gacaggttat caacgaaact tctcagcatc acgatgacct tgaaggggat
     1981 ccaccggtcg ccaccatgag tgtgattaaa ccagacatga agatcaagct gcgtatggaa
     2041 ggcgctgtaa atggacaccc gttcgcgatt gaaggagttg gccttgggaa gcctttcgag
     2101 ggaaaacaga gtatggacct taaagtcata gagggcggac ctctgccttt cgcctatgac
     2161 atcttgacaa ctgcgttctg ttacggcaac agggtattcg ccaaataccc agaaaatata
     2221 gtagactatt tcaagcagtc gtttcctgag ggctactctt gggaacgaag catgaattac
     2281 gaagacgggg gcatttgtaa cgcgacaaac gacataaccc tggatggtga ctgttatatc
     2341 tatgaaattc gatttgatgg tgtgaacttt cctgccaatg gtccagttat gcagaagagg
     2401 actgtgaaat gggagccatc cactgagaaa ttgtatgtgc gtgatggagt gctgaagggc
     2461 gatgttaaca tggctctgtc gcttgaagga ggtggccatt accgatgtga cgtgaaaact
     2521 acttataaag ctaagaaggt tgtccagttg ccagactatc actttgtgga ccaccacatt
     2581 gagattaaaa gccacgacaa agattacagt aatgttaatc tgcatgagca tgccgaagcg
     2641 cattctgagc tgccgaggca ggccaagtaa agcggccgcg actctagatc ataatcagcc
     2701 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2761 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2881 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2941 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3001 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3061 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3121 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3181 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3241 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3301 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3361 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3421 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3481 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3541 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3601 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3781 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3841 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4081 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4681 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4741 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4801 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4861 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4921 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4981 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5041 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5101 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6001 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6061 ttccg
//
