LOCUS       sequence-173988-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of light-regulated p38 inhibitor, firefly 
            luciferase-fused, dark-state mutant photosensor (AsLov2Ja.C450A), 
            inhibitor MK3BD3-13F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Melero-Fernandez de Mera RM, Li LL, Popinigis A, Cisek K, Tuittila 
            M, Yadav L, Serva A, Courtney MJ
  TITLE     A simple optogenetic MAPK inhibitor design reveals resonance between
            transcription-regulating circuitry and temporally-encoded inputs.
  JOURNAL   Nat Commun. 2017 May 12;8:15017. doi: 10.1038/ncomms15017.
  PUBMED    28497795
REFERENCE   2  (bases 1 to 6122)
  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: "Nat 
            Commun."; date: "2017-05-12"; volume: "8:15017. doi"; pages: " 
            10.1038/ncomms15017"
COMMENT     Sequence Label: pLuc-Optop38i-dsm
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..2262
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(679..697)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2910..3031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2947..2966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3001..3020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3038..3493)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3175..3196)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3387..3406
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3520..3624
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3626..3983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3626..3817
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3649..3669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3834..3969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4018..4812
                     /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             4018..4812
                     /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(4072..4091)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4682..4701
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5000..5019)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5044..5091
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5420..6008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5909..5928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg
      661 ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
      721 cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac
      781 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
      841 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
      901 tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac
      961 agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt
     1021 ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg
     1081 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     1141 aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg
     1201 aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc
     1261 gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     1321 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     1381 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc
     1441 cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc
     1501 aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc
     1561 gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc
     1621 aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat
     1681 gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg
     1741 gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg
     1801 attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga
     1861 tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac
     1921 cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc
     1981 atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac
     2041 gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa
     2101 gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt
     2161 gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga
     2221 gagatcctca taaaggccaa gaagggcgga aagatcgccg tgcggtacaa gtccggactc
     2281 agatctcgag ctcaagcttc gaattcactg gcaacaacac tggagagaat cgagaaaaac
     2341 ttcgtcatca ccgaccctag actgcccgat aatcctatca tcttcgcctc cgactcattt
     2401 ctccagctca ctgagtactc tcgggaggaa attctgggcc ggaatgcccg gttcctccag
     2461 ggacctgaaa ccgatagagc cactgtccga aaaatccggg atgccatcga caatcagact
     2521 gaggtgaccg tccagctcat caattacacc aaatctggca aaaaattctg gaacctgttc
     2581 cacctccagc ctatgagaga tcagaagggc gacgtccagt actttattgg cgtccagctc
     2641 gatggaactg aacatgtgag ggacgctgca gaacgagagg gcgtgatgct catcaaaaaa
     2701 accgccgaaa acatcgatga ggccgctaaa gagctgtcta gaaagcggaa gaaggacctg
     2761 cggatctcct gcatgttcta ggatccaccg gatctagata actgatcata atcagccata
     2821 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2881 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2941 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3001 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     3061 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3121 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3181 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3241 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3301 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3361 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3421 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3481 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3541 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3601 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3661 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3841 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3901 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3961 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4021 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4081 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4141 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4201 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4261 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4321 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4381 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4441 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4501 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4561 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4621 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4681 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4741 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4801 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4861 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4921 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4981 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5041 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5101 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5161 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5221 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6121 at
//
