LOCUS       sequence-323619-        5439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 554, Emission: 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5439)
  AUTHORS   Fiolka R, Shao L, Rego EH, Davidson MW, Gustafsson MG
  TITLE     Time-lapse two-color 3D imaging of live cells with doubled 
            resolution using structured illumination.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Apr 3;109(14):5311-5. doi: 
            10.1073/pnas.1119262109. Epub 2012 Mar 19.
  PUBMED    22431626
REFERENCE   2  (bases 1 to 5439)
  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.1119262109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-04-3- 3"; volume: "109"; issue: "14"; pages: "5311-5"
COMMENT     Sequence Label: tdTomato-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..5439
                     /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"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             543..593
                     /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      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             615..2045
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    2167..2288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2204..2223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2258..2277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2295..2750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2432..2453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2644..2663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2777..2881
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2883..3240
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2883..3074
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2906..2926)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3091..3226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3275..4069
                     /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             3275..4069
                     /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(3329..3348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3939..3958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4257..4276)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4301..4348
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4677..5265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5166..5185
                     /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 gctagcgcca
      541 ccatgggcgt ggccgacttg atcaagaagt tcgagtccat ctccaaggag gagggggatc
      601 caccggtcgc caccatggtg agcaagggcg aggaggtcat caaagagttc atgcgcttca
      661 aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
      721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
      781 ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc
      841 accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc
      901 gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg
      961 acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg
     1021 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     1081 gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg
     1141 tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg
     1201 tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1261 agcgctccga gggccgccac cacctgttcc tggggcatgg caccggcagc accggcagcg
     1321 gcagctccgg caccgcctcc tccgaggaca acaacatggc cgtcatcaaa gagttcatgc
     1381 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     1441 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     1501 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg
     1561 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt
     1621 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc
     1681 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg
     1741 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     1801 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact
     1861 acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact
     1921 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     1981 agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac gagctgtaca
     2041 agtaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     2101 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2161 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2221 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2281 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2341 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2401 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2461 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2521 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2581 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2641 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2701 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2761 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2881 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3061 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3241 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3301 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3361 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3421 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3481 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3541 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3601 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3661 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3721 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3781 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3841 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3901 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3961 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4021 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4081 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4141 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4201 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4261 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4321 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4381 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4441 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4501 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4681 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4801 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5281 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5341 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     5401 tacggggtca ttagttcata gcccatatat ggagttccg
//
