LOCUS       sequence-459689-        4222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N-terminally truncated Lifeact fused to 
            C-terminally truncated monomeric superfolder GFP 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 4222)
  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 4222)
  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 msfGFP-Delta-C9-Lifeact-Delta-N2
FEATURES             Location/Qualifiers
     source          1..4222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1010..1131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1047..1066)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1101..1120
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1138..1593)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1275..1296)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1487..1506
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1620..1724
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1726..2083
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1726..1917
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1749..1769)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1934..2069
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2118..2912
                     /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             2118..2912
                     /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(2172..2191)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2782..2801
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3100..3119)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3144..3191
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3520..4108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4009..4028
                     /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 gcatggtgtc caagggcgag gagctgttca ccggcgtggt
      181 gcccatcctg gtggagctgg atggcgacgt gaacggccac aagttcagcg tgcgcggcga
      241 gggcgagggc gacgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      301 gctgcccgtg ccctggccca ccctggtgac caccctgacc tacggcgtgc agtgcttcag
      361 ccgctacccc gatcacatga agcagcacga tttcttcaag agcgccatgc ccgagggcta
      421 cgtgcaggag cgcaccatca gcttcaagga tgacggcacc tacaagaccc gcgccgaggt
      481 gaagttcgag ggcgataccc tggtgaaccg catcgagctg aagggcatcg atttcaagga
      541 ggatggcaac atcctgggcc acaagctgga gtacaacttc aacagccaca acgtgtacat
      601 caccgccgat aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaatgtgga
      661 ggatggctcc gtgcagctgg ccgatcacta ccagcagaac acccccatcg gcgacggccc
      721 agtgctgctg cccgataacc actacctgag cacccagagc aagctgtcca aggaccccaa
      781 cgagaagcgc gatcacatgg tgctgctgga gttcgtgacc gccgccggca tcgtcgcaga
      841 tttgatcaag aaattcgaaa gcatctcaaa ggaagaatga ggatccaccg gatctagata
      901 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
      961 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1021 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1081 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     1141 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1201 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1261 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1321 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1381 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     1441 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     1501 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     1561 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     1621 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     1741 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     1801 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     1861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     1921 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     1981 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2041 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     2101 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2161 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2221 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2281 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2341 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2401 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2461 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2521 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2581 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2641 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2701 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2761 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2821 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2881 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     2941 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3001 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3061 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     3121 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     3181 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3241 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3301 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     3361 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3421 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4141 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4201 taaccgtatt accgccatgc at
//
