LOCUS       sequence-192488-        6119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of light-regulated JNK inhibitor, firefly 
            luciferase-fused, wild-type photosensor, 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
FEATURES             Location/Qualifiers
     source          1..6119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        129..432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        433..636
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     587..607
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      675..684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             681..2330
                     /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(747..765)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2975..3096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3012..3031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3066..3085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3103..3558)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3240..3261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3452..3471
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3585..3689
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3691..4048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3691..3882
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3714..3734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3899..4034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4083..4877
                     /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             4083..4877
                     /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(4137..4156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4747..4766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5065..5084)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5109..5156
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5485..6073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5974..5993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
       61 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      121 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      181 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      241 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      301 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      361 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      421 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      481 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      541 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      601 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      661 tagcgctacc ggtcgccacc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
      721 tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg
      781 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg
      841 ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
      901 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
      961 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     1021 tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc
     1081 aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt
     1141 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     1201 ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac
     1261 tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa
     1321 ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     1381 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     1441 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc
     1501 tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct
     1561 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     1621 ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc
     1681 caggtatcag gcaaggatat gggctcactg agactacatc agctattctg attacacccg
     1741 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     1801 tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag
     1861 gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     1921 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     1981 tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat
     2041 tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg
     2101 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     2161 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     2221 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     2281 aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg cggtacaagt
     2341 ccggactcag atctcgagct caagcttcga attcactggc aacaacactg gagagaatcg
     2401 agaaaaactt cgtcatcacc gaccctagac tgcccgataa tcctatcatc ttcgcctccg
     2461 actcatttct ccagctcact gagtactctc gggaggaaat tctgggccgg aattgccggt
     2521 tcctccaggg acctgaaacc gatagagcca ctgtccgaaa aatccgggat gccatcgaca
     2581 atcagactga ggtgaccgtc cagctcatca attacaccaa atctggcaaa aaattctgga
     2641 acctgttcca cctccagcct atgagagatc agaagggcga cgtccagtac tttattggcg
     2701 tccagctgga tggaactgaa catgtgaggg acgctgcaga acgagagggc gtgatgctca
     2761 tcaaaaaaac cgccgaaaac atcgatgagg ccgctaaaga gctgtctaga agacccaaac
     2821 gaccaaccac actgaacctc ttctaggatc caccggatct agataactga tcataatcag
     2881 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2941 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3001 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3061 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3121 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3181 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3241 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3301 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3361 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3421 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3481 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3541 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3661 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3721 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3781 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3841 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3901 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3961 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4021 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4081 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4261 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4381 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4621 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4861 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4921 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4981 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5041 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5101 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5161 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5221 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5281 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5341 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5401 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5461 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5521 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5581 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5641 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5701 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5761 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5821 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5881 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5941 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6001 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6061 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggc
//
