LOCUS       sequence-425326-        4701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses reversibly switchable fluorescent protein ffDronpa2F 
            targeted to peroxisome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4701)
  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 4701)
  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: pffDronpa2F-peroxisome
FEATURES             Location/Qualifiers
     source          1..4701
                     /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"
     misc_feature    531..611
                     /label=MCS
                     /note="multiple cloning site"
     CDS             619..1290
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVI
                     EGGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNTALSL
                     EGGGHYRCDVKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     polyA_signal    1429..1550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1466..1485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1520..1539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1557..2012)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1694..1715)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1906..1925
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2039..2143
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2145..2502
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2145..2336
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2168..2188)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2353..2488
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2537..3331
                     /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             2537..3331
                     /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(2591..2610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3201..3220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3519..3538)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3563..3610
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3939..4527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4428..4447
                     /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 attctgcagt cgacggtacc gcgggcccgg
      601 gatccaccgg tcgccaccat gagtgtgatt aaaccagaca tgaagatcaa gctgcgtatg
      661 gaaggcgctg taaatggaca cccgttcgcg attgaaggag ttggccttgg gaagcctttc
      721 gagggaaaac agagtatgga ccttaaagtc atagagggcg gacctctgcc tttcgcctat
      781 gacatcttga caactgcgtt ctgttacggc aacagggtat tcgccaaata cccagaaaat
      841 atagtagact atttcaagca gtcgtttcct gagggctact cttgggaacg aagcatgaat
      901 tacgaagacg ggggcatttg taacgcgaca aacgacataa ccctggatgg tgactgttat
      961 atctatgaaa ttcgatttga tggtgtgaac tttcctgcca atggtccagt tatgcagaag
     1021 aggactgtga aatgggagcc atccactgag aaattgtatg tgcgtgatgg agtgctgaag
     1081 ggtgatgtta acacggctct gtcgcttgaa ggaggtggcc attaccgatg tgacgtgaaa
     1141 actacttata aagctaagaa ggttgtccag ttgccagact atcactttgt ggaccaccac
     1201 attgagatta aaagccacga caaagattac agtaatgtta atctgcatga gcatgccgaa
     1261 gcgcattctg agctgccgag gcaggccaag tccggatcca agctgtagcg gccgcgactc
     1321 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1381 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1441 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1501 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     1561 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1621 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1681 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1741 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1801 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1861 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1921 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     1981 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2041 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2101 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2161 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2221 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2281 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2341 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2401 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2461 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2521 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2581 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2641 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2701 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2761 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2821 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2881 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2941 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3001 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3061 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3121 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3181 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3241 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3301 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3361 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3421 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3481 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3541 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     3601 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3661 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3721 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     3781 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3841 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3901 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3961 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4021 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4081 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4141 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4201 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4261 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4321 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4381 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4441 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4501 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4561 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4621 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     4681 tagcccatat atggagttcc g
//
