LOCUS       sequence-372879-        5208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S41E phosphomimetic GFP-tagged human cofilin in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5208)
  AUTHORS   Warner H, Franciosa G, van der Borg G, Coenen B, Faas F, Koenig C, 
            de Boer R, Classens R, Maassen S, Baranov MV, Mahajan S, Dabral D, 
            Bianchi F, van Hilten N, Risselada HJ, Roos WH, Olsen JV, Cano LQ, 
            van den Bogaart G
  TITLE     Atypical cofilin signaling drives dendritic cell migration through 
            the extracellular matrix via nuclear deformation.
  JOURNAL   Cell Rep. 2024 Feb 27;43(3):113866. doi: 
            10.1016/j.celrep.2024.113866.
  PUBMED    38416638
REFERENCE   2  (bases 1 to 5208)
  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: "Cell Rep.";
            date: "2024-02-27"; volume: "43(3):113866. doi"; pages: " 
            10.1016/j.celrep.2024.113866"
COMMENT     Sequence Label: peGFP-N1-Cofilin-S41E
FEATURES             Location/Qualifiers
     source          1..5208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        162..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1249..1258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1255..1257,1258..1260,1261..1974)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1255..1257,1258..1260,1261..1971)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1300..1321)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1561..1580)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1908..1929
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2097..2218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2134..2153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2188..2207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2225..2680)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2362..2383)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2574..2593
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2707..2811
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2813..3170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2813..3004
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2836..2856)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3021..3156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3205..3999
                     /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             3205..3999
                     /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(3259..3278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3869..3888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4187..4206)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4231..4278
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4607..5195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5096..5115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
       61 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca gatctcgagc
      721 tcaagcttcg aattcatggc ctccggtgtg gctgtctctg atggtgtcat caaggtgttc
      781 aacgacatga aggtgcgtaa gtcttcaacg ccagaggagg tgaagaagcg caagaaggcg
      841 gtgctcttct gcctggaaga ggacaagaag aacatcatcc tggaggaggg caaggagatc
      901 ctggtgggcg atgtgggcca gactgtcgac gatccctacg ccacctttgt caagatgctg
      961 ccagataagg actgccgcta tgccctctat gatgcaacct atgagaccaa ggagagcaag
     1021 aaggaggatc tggtgtttat cttctgggcc cccgagtctg cgccccttaa gagcaaaatg
     1081 atttatgcca gctccaagga cgccatcaag aagaagctga cagggatcaa gcatgaattg
     1141 caagcaaact gctacgagga ggtcaaggac cgctgcaccc tggcagagaa gctggggggc
     1201 agtgcggtca tctccctgga gggcaagcct ttgtgggatc caccggtcgc caccatggtg
     1261 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1321 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1381 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1441 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1501 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1561 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1621 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1681 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1741 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1861 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     1981 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2041 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2101 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2161 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     2221 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2281 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2341 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2401 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2461 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2521 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2581 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2641 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2701 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2761 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     2821 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2881 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3001 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3061 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3121 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3181 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3361 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3721 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3841 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4021 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4081 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4141 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4201 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4261 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4321 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4381 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4441 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacg
//
