LOCUS       sequence-459699-        4438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Affimer6 fused to N-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 4438)
  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 4438)
  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 Affimer6-msfGFP-Delta-N12
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1226..1347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1263..1282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1317..1336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1354..1809)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1491..1512)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1703..1722
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1836..1940
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1942..2299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1942..2133
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1965..1985)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2150..2285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2334..3128
                     /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             2334..3128
                     /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(2388..2407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2998..3017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3316..3335)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3360..3407
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3736..4324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4225..4244
                     /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 gcatgaactc cctggaaatc gaagaactgg ctcgtttcgc
      181 tgttgacgaa cacaacaaaa aagaaaacgc tctgctggaa ttcgttcgtg ttgttaaagc
      241 gaaagaacag tcttctgttc cgcattggtg gtggactacc atgtactacc tgaccctgga
      301 agctaaagac ggtggtaaaa agaaactgta cgaagcgaaa gtttgggtta agagagatcc
      361 aaacatgatt ttcaaaatta acttcaaaga actgcaggag ttcaaaccgg ttgtgcccat
      421 cctggtggag ctggatggcg acgtgaacgg ccacaagttc agcgtgcgcg gcgagggcga
      481 gggcgacgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      541 cgtgccctgg cccaccctgg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      601 ccccgatcac atgaagcagc acgatttctt caagagcgcc atgcccgagg gctacgtgca
      661 ggagcgcacc atcagcttca aggatgacgg cacctacaag acccgcgccg aggtgaagtt
      721 cgagggcgat accctggtga accgcatcga gctgaagggc atcgatttca aggaggatgg
      781 caacatcctg ggccacaagc tggagtacaa cttcaacagc cacaacgtgt acatcaccgc
      841 cgataagcag aagaacggca tcaaggccaa cttcaagatc cgccacaatg tggaggatgg
      901 ctccgtgcag ctggccgatc actaccagca gaacaccccc atcggcgacg gcccagtgct
      961 gctgcccgat aaccactacc tgagcaccca gagcaagctg tccaaggacc ccaacgagaa
     1021 gcgcgatcac atggtgctgc tggagttcgt gaccgccgcc ggcatcaccc tgggcatgga
     1081 tgagctgtac aagtgaggat ccaccggatc tagataactg atcataatca gccataccac
     1141 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     1201 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1261 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1321 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     1381 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     1441 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     1501 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1561 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     1621 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     1681 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     1741 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     1801 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1921 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     1981 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2041 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2101 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2161 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2221 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2281 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     2341 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2401 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2461 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     2521 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2581 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2641 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2701 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2761 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2821 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     2881 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     2941 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3001 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3061 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3121 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3181 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3241 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3301 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3361 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     3421 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3481 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3541 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     3601 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3661 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3721 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3781 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3841 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3901 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3961 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4021 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4081 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4141 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4201 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4261 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4321 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4381 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
