LOCUS       sequence-138982-        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 503, Emission: 518.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  AUTHORS   Rego EH, Shao L, Macklin JJ, Winoto L, Johansson GA, Kamps-Hughes N,
            Davidson MW, Gustafsson MG
  TITLE     Nonlinear structured-illumination microscopy with a photoswitchable 
            protein reveals cellular structures at 50-nm resolution.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Jan 17;109(3):E135-43. doi: 
            10.1073/pnas.1107547108. Epub 2011 Dec 12.
  PUBMED    22160683
REFERENCE   2  (bases 1 to 4692)
  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"; doi: 
            "10.1073/pnas.1107547108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-01-17- 17"; volume: "109"; issue: "3"; pages: "E135-43"
COMMENT     Sequence Label: Dronpa-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..4692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             610..660
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      676..685
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..1359
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MVSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKV
                     KEGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNA
                     TNDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALS
                     LEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPR
                     QAK"
     polyA_signal    1487..1608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1524..1543)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1578..1597
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1615..2070)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1752..1773)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1964..1983
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2097..2201
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2203..2560
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2203..2394
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2226..2246)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2411..2546
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2595..3389
                     /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             2595..3389
                     /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(2649..2668)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3259..3278
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3577..3596)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3621..3668
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3997..4585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4486..4505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgccacca tgggcgtggc cgacttgatc aagaagttcg agtccatctc caaggaggag
      661 ggggatccac cggtcgccac catggtgagt gtgattaaac cagacatgaa gatcaagctg
      721 cgtatggaag gcgctgtaaa tggacacccg ttcgcgattg aaggagttgg ccttgggaag
      781 cctttcgagg gaaaacagag tatggacctt aaagtcaaag aaggcggacc tctgcctttc
      841 gcctatgaca tcttgacaac tgtgttctgt tacggcaaca gggtattcgc caaataccca
      901 gaaaatatag tagactattt caagcagtcg tttcctgagg gctactcttg ggaacgaagc
      961 atgaattacg aagacggggg catttgtaac gcgacaaacg acataaccct ggatggtgac
     1021 tgttatatct atgaaattcg atttgatggt gtgaactttc ctgccaatgg tccagttatg
     1081 cagaagagga ctgtgaaatg ggagccatcc actgagaaat tgtatgtgcg tgatggagtg
     1141 ctgaagggtg atgttaacat ggctctgtcg cttgaaggag gtggccatta ccgatgtgac
     1201 ttcaaaacta cttataaagc taagaaggtt gtccagttgc cagactatca ctttgtggac
     1261 caccacattg agattaaaag ccacgacaaa gattacagta atgttaatct gcatgagcac
     1321 gccgaagcgc attctgagct gccgaggcag gccaagtaat gataggcggc cgcgactcta
     1381 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1441 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1501 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1561 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     1621 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1681 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1741 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1801 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1861 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1921 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1981 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2041 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     2101 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2161 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     2221 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2281 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2341 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2401 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2461 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2521 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     2581 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2641 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2701 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2761 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2821 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2881 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2941 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3001 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3061 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3121 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3181 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3241 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3301 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3361 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3421 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3481 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3541 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     3601 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     3661 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3721 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3781 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     3841 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3901 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3961 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4021 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4081 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4141 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4201 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4261 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4321 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4381 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4441 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4501 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4561 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4621 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4681 ccgtattacc gc
//
