LOCUS       sequence-459697-        4399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Affimer6 fused to N-terminally truncated 
            superfolder Cherry2, 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 4399)
  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 4399)
  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-sfCherry2-Delta-N12
FEATURES             Location/Qualifiers
     source          1..4399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(510..528)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1187..1308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1224..1243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1278..1297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1315..1770)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1452..1473)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1664..1683
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1797..1901
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1903..2260
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1903..2094
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1926..1946)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2111..2246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2295..3089
                     /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             2295..3089
                     /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(2349..2368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2959..2978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3277..3296)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3321..3368
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3697..4285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4186..4205
                     /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 ttatgagatt
      421 caaggtgcac atggagggca gcgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      481 gggccacccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
      541 gcccttcgcc tgggacatcc tgagccccca gttcatgtac ggcagcaagg cctacgtgaa
      601 gcaccccgcc gacatccccg actacctgaa gctgagcttc cccgagggct tcacctggga
      661 gagagtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggaca gcagcctgca
      721 ggacggccag ttcatctaca aggtgaagct gctgggcatc aacttcccca gcgacggccc
      781 cgtgatgcag aagaagacca tgggctggga ggccagcacc gagagaatgt accccgagga
      841 cggcgccctg aagggcgaga tcaaccagag actgaagctg aaggacggcg gccactacga
      901 cgccgaggtg aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
      961 cgtggacatc aagctggaca tcaccagcca caacgaggac tacaccatcg tggagcagta
     1021 cgagagagcc gaggccagac acagcaccgg cggctaagga tccaccggat ctagataact
     1081 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1141 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1201 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1261 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     1321 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1381 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1441 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1501 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1561 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1621 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1681 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     1741 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     1801 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1861 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     1921 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     1981 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2041 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2101 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2161 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2221 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     2281 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2341 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2401 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2461 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2521 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2581 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2641 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2701 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     2761 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     2821 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     2881 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     2941 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3001 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3061 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3121 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3181 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3241 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     3301 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     3361 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3421 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3481 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     3541 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3601 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3841 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4321 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4381 ccgtattacc gccatgcat
//
