LOCUS       sequence-192490-        6119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of light-regulated JNK inhibitor, firefly 
            luciferase-fused, lit-state mutant photosensor (AsLov2Ja.I539E), 
            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-lsm
FEATURES             Location/Qualifiers
     source          1..6119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        443..646
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     597..617
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      685..694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             691..2340
                     /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(757..775)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2985..3106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3076..3095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3113..3568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3250..3271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3462..3481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3595..3699
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3701..4058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3701..3892
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3724..3744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3909..4044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4093..4887
                     /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             4093..4887
                     /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(4147..4166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4757..4776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5075..5094)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5119..5166
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5495..6083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5984..6003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
       61 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
      121 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      181 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      241 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      301 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      361 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      421 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      481 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      541 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      601 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      661 tcagatccgc tagcgctacc ggtcgccacc atggaagacg ccaaaaacat aaagaaaggc
      721 ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg
      781 aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac
      841 atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
      901 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
      961 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
     1021 gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa
     1081 aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt
     1141 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
     1201 catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag
     1261 acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg
     1321 cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa
     1381 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     1441 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     1501 gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc
     1561 ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     1621 gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg
     1681 ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc agctattctg
     1741 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     1801 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg
     1861 tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     1921 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa
     1981 cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct
     2041 cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca
     2101 ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     2161 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     2221 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     2281 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg
     2341 cggtacaagt ccggactcag atctcgagct caagcttcga attcactggc aacaacactg
     2401 gagagaatcg agaaaaactt cgtcatcacc gaccctagac tgcccgataa tcctatcatc
     2461 ttcgcctccg actcatttct ccagctcact gagtactctc gggaggaaat tctgggccgg
     2521 aattgccggt tcctccaggg acctgaaacc gatagagcca ctgtccgaaa aatccgggat
     2581 gccatcgaca atcagactga ggtgaccgtc cagctcatca attacaccaa atctggcaaa
     2641 aaattctgga acctgttcca cctccagcct atgagagatc agaagggcga cgtccagtac
     2701 tttattggcg tccagctgga tggaactgaa catgtgaggg acgctgcaga acgagagggc
     2761 gtgatgctca tcaaaaaaac cgccgaaaac gaggacgagg ccgctaaaga gctgtctaga
     2821 agacccaaac gaccaaccac actgaacctc ttctaggatc caccggatct agataactga
     2881 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2941 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3121 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3181 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3241 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3301 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3361 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3421 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3481 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3541 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3601 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3661 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3721 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3781 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3841 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3901 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3961 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4021 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4081 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4141 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4201 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4261 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4321 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4381 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4441 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4501 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4561 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4621 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4681 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4741 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4801 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4861 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4921 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4981 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5041 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5101 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5161 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5221 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5281 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5341 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5401 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5461 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5521 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5581 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5641 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5701 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5761 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5821 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5881 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5941 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6001 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6061 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcc
//
