LOCUS       sequence-319424-        5820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a control for optogenetic cofilin analog.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   Stone OJ, Pankow N, Liu B, Sharma VP, Eddy RJ, Wang H, Putz AT, 
            Teets FD, Kuhlman B, Condeelis JS, Hahn KM
  TITLE     Optogenetic control of cofilin and alphaTAT in living cells using 
            Z-lock.
  JOURNAL   Nat Chem Biol. 2019 Dec;15(12):1183-1190. doi: 
            10.1038/s41589-019-0405-4. Epub 2019 Nov 18.
  PUBMED    31740825
REFERENCE   2  (bases 1 to 5820)
  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"; doi: 
            "10.1038/s41589-019-0405-4"; journalName: "Nat Chem Biol"; date: 
            "2019-12"; volume: "15"; issue: "12"; pages: "1183-1190"
COMMENT     Sequence Label: Mammalian expression: Z-lock cofilin dark stat...
FEATURES             Location/Qualifiers
     source          1..5820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             564..1271
                     /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(709..727)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     977..996
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1287..1460
                     /codon_start=1
                     /product="""Zdark"" protein, derived from the Z subunit of 
                     protein A, that binds to the dark state of LOV2 and is 
                     released upon irradiation (Wang et al., 2016)"
                     /label=Zdk2
                     /translation="VDNKFNKEMLSARVEIYGLPNLNWGQRFAFFSSLTDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1998..2423
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="ATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGRN
                     ARFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQY
                     FIGVQKDGTEHVRDAAEREAVMEIKKTAEEIDEAAKEL"
     polyA_signal    2548..2669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2585..2604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2639..2658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2676..3131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2813..2834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3025..3044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3158..3262
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3264..3621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3264..3455
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3287..3307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3472..3607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3656..4450
                     /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             3656..4450
                     /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(3710..3729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4320..4339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4638..4657)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4682..4729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5058..5646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5547..5566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagcc accatggtga gcaagggcga ggaggataac atggccatca
      601 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      661 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      721 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      781 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
      841 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
      901 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
      961 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1021 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1081 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1141 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1201 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1261 agctgtacaa gggaagcggt agtggagtgg ataacaaatt caataaagaa atgttgagtg
     1321 cccgtgtgga aatctatggt ctgccaaacc tgaattgggg tcagaggttt gccttcttca
     1381 gtagcctgac ggatgatcca tctcagagcg ccaatctgct ggccgaagcc aaaaaactga
     1441 acgatgccca ggccccaaaa ggttctggag gcggaatggc ctctggtgtg gctgtctctg
     1501 atggagtcat caaggtgttc aatgacatga aagttcgcaa gtcttcaacg ccagaagaag
     1561 tgaagaaacg caagaaggca gtgctctttt gcctgagtga ggacaagaag aacatcatcc
     1621 tggaggaggg caaggagatt ctggtaggag atgtggggca gactgtggac gacccctaca
     1681 ccacttttgt caagatgctg ccagacaagg actgccgcta tgctctctat gacgcaacct
     1741 atgagaccaa ggagagcaag aaggaggacc tggtattcat tttctgggcc cccgagagtg
     1801 caccccttaa gagcaaaatg atctatgcca gctccaagga tgccatcaag aagaaactga
     1861 caggaatcaa gcacgaatta caagctaact gctacgagga ggtcaaggac cgctgcaccc
     1921 tggcagagaa actaggtggc agcgccgtca tttccctgga gggcaagcct ttgggcggta
     1981 gcggagggag cggcggtgct actacacttg aacgtattga gaagaacttt gtcattactg
     2041 acccaagatt gccagataat cccattatat tcgcgtccga tagtttcttg cagttgacag
     2101 aatatagccg tgaagaaatt ttgggaagaa acgccaggtt tctacaaggt cctgaaactg
     2161 atcgcgcgac agtgagaaaa attagagatg ccatagataa ccaaacagag gtcactgttc
     2221 agctgattaa ttatacaaag agtggtaaaa agttctggaa cctctttcac ttgcagccta
     2281 tgcgagatca gaagggagat gtccagtact ttattggggt tcagaaggat ggaactgagc
     2341 atgtccgaga tgctgccgag agagaggcag tcatggagat taagaaaact gcagaagaaa
     2401 ttgatgaggc ggcaaaagaa ctttaagcgg ccgcgactct agatcataat cagccatacc
     2461 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2521 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2581 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2641 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2701 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2761 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2821 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2881 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2941 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3001 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3061 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3121 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3241 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3301 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3421 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3481 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3541 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3601 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3661 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3721 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3781 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3841 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3901 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3961 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4021 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4081 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4141 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4201 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4261 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4321 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4381 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4441 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4501 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4561 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4621 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4681 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4741 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4801 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4861 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4981 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     5761 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
