LOCUS       sequence-416460-        6884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ActuAtor for iLID optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6884)
  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 6884)
  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-mSspB-mCherry
FEATURES             Location/Qualifiers
     source          1..6884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        85..442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        85..276
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(108..128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      293..428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             477..1271
                     /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             477..1271
                     /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(531..550)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1141..1160
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1459..1478)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1503..1550
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1879..2467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2368..2387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2642..2945
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2946..3149
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3100..3120
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3200..3209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5414..5423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5420..6130
                     /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(5565..5583)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5833..5852
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    6253..6374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6290..6309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6344..6363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6381..6836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6518..6539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6730..6749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6863..6884,1..83)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
       61 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
      121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      361 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
      421 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
      481 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      541 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      601 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
      661 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
      721 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
      781 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
      841 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
      901 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
      961 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     1021 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1081 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     1141 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     1201 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     1261 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     1321 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1381 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1441 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     1501 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     1561 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     1621 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     1681 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     1741 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1801 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1861 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1921 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1981 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2041 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2101 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2161 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2221 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2281 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2341 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2401 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2461 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2521 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     2581 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     2641 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     2701 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     2761 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     2821 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     2881 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     2941 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     3001 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     3061 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     3121 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     3181 accggactca gatctcgagg ccaccatggc tacagattca gaggatagct ccctgaacac
     3241 agacgagtgg gaggaggaga agaccgagga gcagccctcc gaagtgaata caggccctag
     3301 atacgagacc gccagagagg tgtctagccg ggacatcaag gagctggaga agtctaacaa
     3361 ggtgcgcaac acaaataagg ccgatctgat cgccatgctg aaggagaagg ccgagaaggg
     3421 cccaaatatc aacaataaca attccgagca gaccgagaac gccgccatca atgaggaggc
     3481 ctctggagca gacaggccag caatccaggt ggagaggaga cacccaggac tgccatccga
     3541 ttctgccgca gagatcaaga agcggcgcaa ggccatcgcc tcctctgact ctgagctgga
     3601 gagcctgaca tatcctgata agccaaccaa ggtgaacaag aagaaggtgg caaaggagag
     3661 cgtggcagac gccagcgagt ccgacctgga tagctccatg cagtccgccg atgagtctag
     3721 cccacagccc ctgaaggcca accagcagcc cttctttcct aaggtgttca agaagatcaa
     3781 ggacgccggc aagtgggtgc gcgacaagat cgatgagaat cccgaggtga agaaggccat
     3841 cgtggacaag agcgccggcc tgatcgatca gctgctgacc aagaagaaga gcgaggaggt
     3901 gaacgccagc gacttccccc ctccacccac agatgaggag ctgagactgg ccctgccaga
     3961 gaccccaatg ctgctgggct ttaatgcccc agccacaagc gagccctcct ctttcgagtt
     4021 tcctccacca cctactgacg aggagctgcg gctggccctg cctgagacac caatgctgct
     4081 gggcttcaac gcacctgcca cctccgagcc aagctccttc gagtttccac cccctccaac
     4141 cgaggatgag ctggagatca tcagagagac agcctctagc ctggactcct cttttacccg
     4201 gggcgatctg gcctccctga ggaacgccat caataggcac tcccagaact tctctgactt
     4261 tccccctatc cctacagagg aggagctgaa tggaaggggc ggcaggccca ccagcgagga
     4321 gttcagctcc ctgaactctg gcgactttac agacgatgag aacagcgaga ccacagagga
     4381 ggagatcgat cgcctggccg acctgaggga tagaggcacc ggcaagcaca gcaggaacgc
     4441 aggcttcctg cccctgaatc cttttgcctc tagccctgtg ccatctctga gcccaaaggt
     4501 gagcaagatc agcgcccccg ccctgatcag cgatatcaca aagaagaccc ctttcaagaa
     4561 cccatcccag cccctgaacg tgtttaataa gaagaccaca accaagacag tgaccaagaa
     4621 gcccacacct gtgaagaccg caccaaagct ggccgagctg ccagccacaa agcctcagga
     4681 gaccgtgctg agagagaata agacaccttt catcgagaag caggccgaga ccaacaagca
     4741 gagcatcaat atgccttccc tgccagtgat ccagaaggag gccacagaga gcgacaagga
     4801 ggagatgaag cctcagaccg aggagaagat ggtggaggag tccgagtctg ccaacaatgc
     4861 caacggcaag aatcggtctg ccggcatcga ggagggcaag ctgatcgcca agagcgccga
     4921 ggatgagaag gccaaggagg agccaggcaa ccacacaacc cgaattctgc agtcgacggt
     4981 accgcgggcc cgggatccaa gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc
     5041 tggtggtagt gctggtggta gctccccaaa gaggcctaag ctgctgagag agtactatga
     5101 ctggctggtg gataacagct tcacccccta cctggtggtg gacgccacat atctgggcgt
     5161 gaacgtgcct gtggagtacg tgaaggatgg ccagatcgtg ctgaatctga gcgcctccgc
     5221 caccggaaac ctgcagctga caaatgactt catccagttt aatgcccagt tcaagggcgt
     5281 gtccagggag ctctacatcc caatgggagc cgccctggca atctatgcaa gggagaacgg
     5341 cgacggcgtg atgtttgagc ctgaagaaat ctacgatgag ctgaatatcg gcagtgctgg
     5401 tggtccaccg gtcgccacca tggtaagcaa gggcgaggag gataacatgg ccatcatcaa
     5461 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     5521 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     5581 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     5641 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     5701 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     5761 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     5821 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     5881 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     5941 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     6001 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     6061 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     6121 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     6181 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     6241 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6301 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6361 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     6421 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6481 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6541 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6601 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6661 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6721 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6781 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     6841 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttat
//
