LOCUS       sequence-192487-        6119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of light-regulated JNK inhibitor, firefly 
            luciferase-fused, dark-state mutant photosensor (AsLov2Ja.C450A), 
            inhibitor JIP11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6119)
  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 6119)
  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-OptoJNKi-dsm
FEATURES             Location/Qualifiers
     source          1..6119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        151..454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        455..658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     609..629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      697..706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             703..2352
                     /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(769..787)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2997..3118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3034..3053)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3088..3107
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3125..3580)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3262..3283)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3474..3493
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3607..3711
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3713..4070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3713..3904
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3736..3756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3921..4056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4105..4899
                     /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             4105..4899
                     /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(4159..4178)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4769..4788
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5087..5106)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5131..5178
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5507..6095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5996..6015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
       61 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
      121 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      181 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      241 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      361 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      421 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      481 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      541 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      601 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      661 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggaaga cgccaaaaac
      721 ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg
      781 cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat
      841 atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct
      901 atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt
      961 caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac
     1021 gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg
     1081 ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc
     1141 caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg
     1201 ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc
     1261 gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa
     1321 ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt
     1381 tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt
     1441 tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat
     1501 agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg
     1561 ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta
     1621 tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg
     1681 gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca
     1741 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
     1801 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa
     1861 agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa
     1921 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     1981 gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc
     2041 tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac
     2101 gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     2161 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     2221 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     2281 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     2341 aagatcgccg tgcggtacaa gtccggactc agatctcgag ctcaagcttc gaattcactg
     2401 gcaacaacac tggagagaat cgagaaaaac ttcgtcatca ccgaccctag actgcccgat
     2461 aatcctatca tcttcgcctc cgactcattt ctccagctca ctgagtactc tcgggaggaa
     2521 attctgggcc ggaatgcccg gttcctccag ggacctgaaa ccgatagagc cactgtccga
     2581 aaaatccggg atgccatcga caatcagact gaggtgaccg tccagctcat caattacacc
     2641 aaatctggca aaaaattctg gaacctgttc cacctccagc ctatgagaga tcagaagggc
     2701 gacgtccagt actttattgg cgtccagctc gatggaactg aacatgtgag ggacgctgca
     2761 gaacgagagg gcgtgatgct catcaaaaaa accgccgaaa acatcgatga ggccgctaaa
     2821 gagctgtcta gaagacccaa acgaccaacc acactgaacc tcttctagga tccaccggat
     2881 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2941 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3001 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3061 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     3121 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3181 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3241 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3301 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3361 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3421 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3481 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3541 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3601 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3661 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3721 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3841 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3901 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3961 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4021 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4081 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4141 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4201 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4261 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4321 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4381 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4441 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4501 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4561 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4621 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4681 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4741 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4801 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4861 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4921 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5041 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5101 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5161 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5221 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5281 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5341 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5401 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5461 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5521 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5581 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5641 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5701 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5761 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5821 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5881 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5941 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6001 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6061 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gccttttta
//
