LOCUS       sequence-372883-        5199 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses S41E phosphomimetic mCherry-tagged human cofilin in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5199)
  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 5199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pmCherry-N1-Cofilin-S41E
FEATURES             Location/Qualifiers
     source          1..5199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        352..555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     506..526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1135..1144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1141..1851
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1286..1304)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1554..1573
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1974..2095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2011..2030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2065..2084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2102..2557)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2239..2260)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2451..2470
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2584..2688
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2690..3047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2690..2881
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2713..2733)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2898..3033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3082..3876
                     /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             3082..3876
                     /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(3136..3155)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3746..3765
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4064..4083)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4108..4155
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4484..5072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4973..4992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
       61 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      121 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      181 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      241 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      301 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      361 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      421 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      481 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      541 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
      601 tcgagctcaa gcttcgaatt catggcctcc ggtgtggctg tctctgatgg tgtcatcaag
      661 gtgttcaacg acatgaaggt gcgtaagtct tcaacgccag aggaggtgaa gaagcgcaag
      721 aaggcggtgc tcttctgcct ggaagaggac aagaagaaca tcatcctgga ggagggcaag
      781 gagatcctgg tgggcgatgt gggccagact gtcgacgatc cctacgccac ctttgtcaag
      841 atgctgccag ataaggactg ccgctatgcc ctctatgatg caacctatga gaccaaggag
      901 agcaagaagg aggatctggt gtttatcttc tgggcccccg agtctgcgcc ccttaagagc
      961 aaaatgattt atgccagctc caaggacgcc atcaagaaga agctgacagg gatcaagcat
     1021 gaattgcaag caaactgcta cgaggaggtc aaggaccgct gcaccctggc agagaagctg
     1081 gggggcagtg cggtcatctc cctggagggc aagcctttgt gggatccacc ggtcgccacc
     1141 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     1201 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1261 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     1321 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     1381 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     1441 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     1501 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1561 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1621 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1681 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1741 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1801 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta aagcggccgc
     1861 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1921 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1981 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2041 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2101 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2161 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2221 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2281 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2341 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2401 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2461 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2521 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2581 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2641 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2701 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2821 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2881 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2941 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3001 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3061 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3121 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3181 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3241 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3301 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3361 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3421 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3481 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3541 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3601 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3661 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3721 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3781 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3841 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3901 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     3961 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4021 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4081 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4141 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4201 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4261 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4321 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4381 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4441 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4501 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4561 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4621 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4681 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4741 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4801 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4861 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4921 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4981 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5041 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5101 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5161 tggataaccg tattaccgcc atgcattagt tattaatag
//
