LOCUS       sequence-459702-        5275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human beta-actin fused intramolecularly to 
            N- and C-terminally truncated monomeric sfGFP, for actin filament 
            organization measurements using fluorescence polarization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5275)
  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 5275)
  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 
            beta-actinh7TAA-msfGFP-Delta-N7-Delta-C11-SSSSactin
FEATURES             Location/Qualifiers
     source          1..5275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2063..2184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2100..2119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2154..2173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2191..2646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2328..2349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2540..2559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2673..2777
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2779..3136
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2779..2970
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2802..2822)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2987..3122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3171..3965
                     /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             3171..3965
                     /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(3225..3244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3835..3854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4153..4172)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4197..4244
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4573..5161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5062..5081
                     /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 gcatggatga tgatatcgcc gcgctcgtcg tcgacaacgg
      181 ctccggcatg tgcaaggccg gcttcgcggg cgacgatgcc ccccgggccg tcttcccctc
      241 catcgtgggg cgccccaggc accagggcgt gatggtgggc atgggtcaga aggattccta
      301 tgtgggcgac gaggcccaga gcaagagagg catcctcacc ctgaagtacc ccatcgagca
      361 cggcatcgtc accaactggg acgacatgga gaaaatctgg caccacacct tctacaatga
      421 gctgcgtgtg gctcccgagg agcaccccgt gctgctgacc gaggcccccc tgaaccccaa
      481 ggccaaccgc gagaagatga cccagatcat gtttgagacc ttcaacaccc cagccatgta
      541 cgttgctatc caggctgtgc tatccctgta cgcctctggc cgtaccactg gcatcgtgat
      601 ggactccggt gacggggtca cccacactgt gcccatctac gaggggtatg ccctccccca
      661 tgccatcctg cgtctggacc tggctggccg ggacctgact gactacctca tgaagatcct
      721 caccgagcgc ggctacagct tcaccaccac ggccgagcgg gaaatcgtgc gtgacattaa
      781 ggagaagctg tgctacgtcg ccctggactt cgagcaagag atggccacgg ctgctctgtt
      841 caccggcgtg gtgcccatcc tggtggagct ggatggcgac gtgaacggcc acaagttcag
      901 cgtgcgcggc gagggcgagg gcgacgccac caacggcaag ctgaccctga agttcatctg
      961 caccaccggc aagctgcccg tgccctggcc caccctggtg accaccctga cctacggcgt
     1021 gcagtgcttc agccgctacc ccgatcacat gaagcagcac gatttcttca agagcgccat
     1081 gcccgagggc tacgtgcagg agcgcaccat cagcttcaag gatgacggca cctacaagac
     1141 ccgcgccgag gtgaagttcg agggcgatac cctggtgaac cgcatcgagc tgaagggcat
     1201 cgatttcaag gaggatggca acatcctggg ccacaagctg gagtacaact tcaacagcca
     1261 caacgtgtac atcaccgccg ataagcagaa gaacggcatc aaggccaact tcaagatccg
     1321 ccacaatgtg gaggatggct ccgtgcagct ggccgatcac taccagcaga acacccccat
     1381 cggcgacggc ccagtgctgc tgcccgataa ccactacctg agcacccaga gcaagctgtc
     1441 caaggacccc aacgagaagc gcgatcacat ggtgctgctg gagttcgtga ccgccgcctc
     1501 cagctcctcc ctggagaaga gctacgagct gcctgacggc caggtcatca ccattggcaa
     1561 tgagcggttc cgctgccctg aggcactctt ccagccttcc ttcctgggca tggagtcctg
     1621 tggcatccac gaaactacct tcaactccat catgaagtgt gacgtggaca tccgcaaaga
     1681 cctgtacgcc aacacagtgc tgtctggcgg caccaccatg taccctggca ttgccgacag
     1741 gatgcagaag gagatcactg ccctggcacc cagcacaatg aagatcaaga tcattgctcc
     1801 tcctgagcgc aagtactccg tgtggatcgg cggctccatc ctggcctcgc tgtccacctt
     1861 ccagcagatg tggatcagca agcaggagta tgacgagtcc ggcccctcca tcgtccaccg
     1921 caaatgcttc tgaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     1981 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2041 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2101 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2161 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     2221 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2281 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2341 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2401 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2461 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2521 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2581 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2641 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2761 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2821 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2881 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2941 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3001 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3061 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3121 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3181 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3241 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3301 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3361 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3421 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3481 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3541 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3601 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3661 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3721 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3781 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3841 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3901 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3961 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4021 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4081 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4141 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4201 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4261 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4321 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4381 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4441 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4501 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4561 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4621 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4681 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4741 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4801 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4861 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4921 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4981 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5041 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5101 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5161 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5221 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
