LOCUS       sequence-459698-        4759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N- and C-terminally truncated Utrophin calponin homology 
            domain fused to C-terminally truncated msfGFP 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 4759)
  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 4759)
  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-C10-Utr28-222
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1547..1668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1584..1603)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1638..1657
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1675..2130)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1812..1833)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2024..2043
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2157..2261
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2263..2620
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2263..2454
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2286..2306)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2471..2606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2655..3449
                     /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             2655..3449
                     /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(2709..2728)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3319..3338
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3637..3656)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3681..3728
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4057..4645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4546..4565
                     /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 gccgccggcg aacacaatga
      841 cgtacagaag aaaaccttta ccaaatggat aaatgctcga ttttcaaaga gtgggaaacc
      901 acccatcaat gatatgttca cagacctcaa agatggaagg aagctattgg atcttctaga
      961 aggcctcaca ggaacatcac tgccaaagga acgtggttcc acaagggtac atgccttaaa
     1021 taacgtcaac agagtgctgc aggttttaca tcagaacaat gtggaattag tgaatatagg
     1081 gggaaccgac attgtggatg gaaatcacaa actgactttg gggttacttt ggagcatcat
     1141 tttgcactgg caggtgaaag atgtcatgaa ggatgtcatg tcggacctgc agcagacgaa
     1201 cagtgagaag atcctgctca gctgggtgcg tcagaccacc aggccctaca gccaagtcaa
     1261 cgtcctcaac ttcaccacca gctggacaga tggactcgcc tttaatgctg tcctccaccg
     1321 acataaacct gatctcttca gctgggataa agttgtcaaa atgtcaccaa ttgagagact
     1381 tgaacatgcc ttcagcaagg ctcaaactta tttgtgagga tccaccggat ctagataact
     1441 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1501 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1561 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1621 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     1681 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1741 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1801 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1861 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1921 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1981 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2041 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2101 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     2161 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2221 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     2281 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2341 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2401 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2461 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2521 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2581 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     2641 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2701 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2761 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2821 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2881 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2941 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3001 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3061 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3121 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3181 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3241 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3301 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3361 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3421 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3481 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3541 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3601 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     3661 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     3721 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3781 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3841 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     3901 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3961 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4021 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4081 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4141 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4201 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4261 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4321 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4381 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4441 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4501 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4561 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4621 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4681 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4741 ccgtattacc gccatgcat
//
