LOCUS       sequence-459701-        4279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of C-terminally truncated F-tractin fused to 
            N-terminally truncated monomeric sfGFP, for actin filament 
            organization measurements using fluorescence polarization 
            microscopy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4279)
  AUTHORS   Martins CS, Iv F, Suman SK, Panagiotou TC, Sidor C, Ruso-Lopez M, 
            Plancke CN, Omi S, Pages R, Gomes M, Llewellyn A, Bandi SR, Ramond 
            L, Arbizzani F, Rimoli CV, Schnorrer F, Robin F, Wilde A, LeGoff L, 
            Pedelacq JD, Jegou A, Cabantous S, Rincon SA, Chandre C, Brasselet 
            S, Mavrakis M
  TITLE     Genetically encoded reporters of actin filament organization in 
            living cells and tissues.
  JOURNAL   Cell. 2025 Apr 1:S0092-8674(25)00276-4. doi: 
            10.1016/j.cell.2025.03.003.
  PUBMED    40179884
REFERENCE   2  (bases 1 to 4279)
  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. 2025 
            Apr 1:S0092-8674(25)00276-4. doi: 10.1016/j.cell.2025.03.003."
COMMENT     Sequence Label: pCMVtrunc F-tractinN9-39-msfGFP-Delta-N7
FEATURES             Location/Qualifiers
     source          1..4279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1067..1188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1104..1123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1158..1177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1195..1650)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1332..1353)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1544..1563
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1677..1781
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1783..2140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1783..1974
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1806..1826)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1991..2126
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2175..2969
                     /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             2175..2969
                     /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(2229..2248)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2839..2858
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3157..3176)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3201..3248
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3577..4165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4066..4085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tgggaggtct atataagcag agctggttta
      121 gtgaaccgtc agatccgcta gcatgggcat ggcgcgacca cggggcgcgg ggccctgcag
      181 ccccgggttg gagcgggctc cgcgccggag cgtcggggag ctgcgcctgc tcttcgaact
      241 gttcaccggc gtggtgccca tcctggtgga gctggatggc gacgtgaacg gccacaagtt
      301 cagcgtgcgc ggcgagggcg agggcgacgc caccaacggc aagctgaccc tgaagttcat
      361 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctg gtgaccaccc tgacctacgg
      421 cgtgcagtgc ttcagccgct accccgatca catgaagcag cacgatttct tcaagagcgc
      481 catgcccgag ggctacgtgc aggagcgcac catcagcttc aaggatgacg gcacctacaa
      541 gacccgcgcc gaggtgaagt tcgagggcga taccctggtg aaccgcatcg agctgaaggg
      601 catcgatttc aaggaggatg gcaacatcct gggccacaag ctggagtaca acttcaacag
      661 ccacaacgtg tacatcaccg ccgataagca gaagaacggc atcaaggcca acttcaagat
      721 ccgccacaat gtggaggatg gctccgtgca gctggccgat cactaccagc agaacacccc
      781 catcggcgac ggcccagtgc tgctgcccga taaccactac ctgagcaccc agagcaagct
      841 gtccaaggac cccaacgaga agcgcgatca catggtgctg ctggagttcg tgaccgccgc
      901 cggcatcacc ctgggcatgg atgagctgta caagtgagga tccaccggat ctagataact
      961 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1021 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     1201 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1261 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1321 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1381 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1441 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1501 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1561 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     1621 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     1681 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1741 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     1801 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     1861 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1921 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     1981 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2041 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2101 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     2161 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2221 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2281 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2341 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2401 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2461 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2521 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2581 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     2641 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     2701 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     2761 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     2821 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     2881 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     2941 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3001 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3061 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3121 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     3181 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     3241 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3301 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3361 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     3421 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3481 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3541 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3601 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3661 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3721 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3781 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3841 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3901 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3961 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4021 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4081 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4141 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4201 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4261 ccgtattacc gccatgcat
//
