LOCUS       sequence-416399-        6872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ActuAtor for FKBP-FRB CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6872)
  AUTHORS   Nakamura H, Rho E, Lee CT, Itoh K, Deng D, Watanabe S, Razavi S, 
            Matsubayashi HT, Zhu C, Jung E, Rangamani P, Watanabe S, Inoue T
  TITLE     ActuAtor, a Listeria-inspired molecular tool for physical 
            manipulation of intracellular organizations through de novo actin 
            polymerization.
  JOURNAL   Cell Rep. 2023 Sep 2:113089. doi: 10.1016/j.celrep.2023.113089.
  PUBMED    37734382
REFERENCE   2  (bases 1 to 6872)
  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 Rep. 
            2023 Sep 2:113089. doi: 10.1016/j.celrep.2023.113089."
COMMENT     Sequence Label: mActA-FKBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1544..1864
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     regulatory      1886..1895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1892..2602
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2037..2055)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2305..2324
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2725..2846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2762..2781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2816..2835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2853..3308)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2990..3011)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3202..3221
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3335..3439
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3441..3798
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3441..3632
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3464..3484)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3649..3784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3833..4627
                     /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             3833..4627
                     /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(3887..3906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4497..4516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4815..4834)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4859..4906
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5235..5823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5724..5743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5998..6301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6302..6505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6456..6476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6556..6565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gagacaccca ggactgccat ccgattctgc cgcagagatc aagaagcggc gcaaggccat
       61 cgcctcctct gactctgagc tggagagcct gacatatcct gataagccaa ccaaggtgaa
      121 caagaagaag gtggcaaagg agagcgtggc agacgccagc gagtccgacc tggatagctc
      181 catgcagtcc gccgatgagt ctagcccaca gcccctgaag gccaaccagc agcccttctt
      241 tcctaaggtg ttcaagaaga tcaaggacgc cggcaagtgg gtgcgcgaca agatcgatga
      301 gaatcccgag gtgaagaagg ccatcgtgga caagagcgcc ggcctgatcg atcagctgct
      361 gaccaagaag aagagcgagg aggtgaacgc cagcgacttc ccccctccac ccacagatga
      421 ggagctgaga ctggccctgc cagagacccc aatgctgctg ggctttaatg ccccagccac
      481 aagcgagccc tcctctttcg agtttcctcc accacctact gacgaggagc tgcggctggc
      541 cctgcctgag acaccaatgc tgctgggctt caacgcacct gccacctccg agccaagctc
      601 cttcgagttt ccaccccctc caaccgagga tgagctggag atcatcagag agacagcctc
      661 tagcctggac tcctctttta cccggggcga tctggcctcc ctgaggaacg ccatcaatag
      721 gcactcccag aacttctctg actttccccc tatccctaca gaggaggagc tgaatggaag
      781 gggcggcagg cccaccagcg aggagttcag ctccctgaac tctggcgact ttacagacga
      841 tgagaacagc gagaccacag aggaggagat cgatcgcctg gccgacctga gggatagagg
      901 caccggcaag cacagcagga acgcaggctt cctgcccctg aatccttttg cctctagccc
      961 tgtgccatct ctgagcccaa aggtgagcaa gatcagcgcc cccgccctga tcagcgatat
     1021 cacaaagaag acccctttca agaacccatc ccagcccctg aacgtgttta ataagaagac
     1081 cacaaccaag acagtgacca agaagcccac acctgtgaag accgcaccaa agctggccga
     1141 gctgccagcc acaaagcctc aggagaccgt gctgagagag aataagacac ctttcatcga
     1201 gaagcaggcc gagaccaaca agcagagcat caatatgcct tccctgccag tgatccagaa
     1261 ggaggccaca gagagcgaca aggaggagat gaagcctcag accgaggaga agatggtgga
     1321 ggagtccgag tctgccaaca atgccaacgg caagaatcgg tctgccggca tcgaggaggg
     1381 caagctgatc gccaagagcg ccgaggatga gaaggccaag gaggagccag gcaaccacac
     1441 aacccgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaagtgctg gtggtagtgc
     1501 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtggagtgc aggtggaaac
     1561 catctcccca ggagacgggc gcaccttccc caagcgcggc cagacctgcg tggtgcacta
     1621 caccgggatg cttgaagatg gaaagaaatt tgattcctcc cgggacagaa acaagccctt
     1681 taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat
     1741 gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg
     1801 gcacccaggc atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaact
     1861 ggaaagtgct ggtggtccac cggtcgccac catggtaagc aagggcgagg aggataacat
     1921 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     1981 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2041 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2101 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2161 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2221 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2281 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2341 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2401 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2461 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2521 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2581 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     2641 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2701 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2761 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2821 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     2881 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2941 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3001 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3061 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3121 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3181 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3241 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3301 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3361 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3421 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3481 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3541 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3601 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3661 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3721 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3781 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3841 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3901 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3961 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4021 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4081 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4141 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4201 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4261 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4321 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4381 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4441 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4501 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4561 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4621 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4681 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4741 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4801 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4861 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     4921 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4981 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5041 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5101 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5161 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5221 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5281 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5341 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5401 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5461 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5521 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5581 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5641 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5701 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5761 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5821 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5881 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     5941 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     6001 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     6061 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6121 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6181 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     6241 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6301 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     6361 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     6421 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     6481 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     6541 gactcagatc tcgaggccac catggctaca gattcagagg atagctccct gaacacagac
     6601 gagtgggagg aggagaagac cgaggagcag ccctccgaag tgaatacagg ccctagatac
     6661 gagaccgcca gagaggtgtc tagccgggac atcaaggagc tggagaagtc taacaaggtg
     6721 cgcaacacaa ataaggccga tctgatcgcc atgctgaagg agaaggccga gaagggccca
     6781 aatatcaaca ataacaattc cgagcagacc gagaacgccg ccatcaatga ggaggcctct
     6841 ggagcagaca ggccagcaat ccaggtggag ag
//
