LOCUS       sequence-194847-        6723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of light-regulated JNK inhibitor, mCherry-tagged, 
            localised to chromatin with H2B, wild-type photosensor, inhibitor 
            JIP11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6723)
  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 6723)
  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: pKCAG-H2BmCherry-OptoJNKi
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..642
                     /label=chicken beta-actin promoter
     intron          643..1651
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1575..1597
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1772..2149
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      2162..2171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2168..2875
                     /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(2313..2331)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2581..2600
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3511..3632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3548..3567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3602..3621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3639..4094)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3776..3797)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3988..4007
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4121..4225
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4227..4584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4227..4418
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4250..4270)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4435..4570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4619..5413
                     /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             4619..5413
                     /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(4673..4692)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5283..5302
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5601..5620)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5645..5692
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6021..6609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6510..6529
                     /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 atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catgggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      421 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      481 ggggggcgcg cgccaggcgg ggcggggcgg gggcgagggc ggggcggggc gaggcggaga
      541 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      601 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgttgcct
      661 tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      721 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      781 gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa gccttaaagg gctccgggag
      841 ggccctttgt gcggggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
      901 cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      961 tgtgcgctcc gcgtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     1021 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     1081 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc
     1141 acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg
     1201 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     1261 gctcggggga ggggcgcggc ggccccggag cgccggcggc tgtcgaggcg cggcgagccg
     1321 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     1381 ctggcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cgggcgaagc
     1441 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     1501 gtccccttct ccatctccag cctcggggct gccgcagggg gacggctgcc ttcggggggg
     1561 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
     1621 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttgttgtgct
     1681 gtctcatcat tttggcaaag gaattgctag cgctaccgga ctcagatctc gagctcaagc
     1741 ttcgaattct gcagtcgacg gtaccgccac catgccagag ccagcgaagt ctgctcccgc
     1801 cccgaaaaag ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg
     1861 caagcgcagc cgcaaggaga gctattccat ctatgtgtac aaggttctga agcaggtcca
     1921 ccctgacacc ggcatttcgt ccaaggccat gggcatcatg aattcgtttg tgaacgacat
     1981 tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat tacaacaagc gctcgaccat
     2041 cacctccagg gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca
     2101 cgccgtgtcc gagggtacta aggccatcac caagtacacc agcgctaagg atccaccggt
     2161 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2221 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2281 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2341 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2401 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2461 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2521 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2581 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2641 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2701 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2761 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2821 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtccgg
     2881 actcagatct cgagctcaag cttcgaattc actggcaaca acactggaga gaatcgagaa
     2941 aaacttcgtc atcaccgacc ctagactgcc cgataatcct atcatcttcg cctccgactc
     3001 atttctccag ctcactgagt actctcggga ggaaattctg ggccggaatt gccggttcct
     3061 ccagggacct gaaaccgata gagccactgt ccgaaaaatc cgggatgcca tcgacaatca
     3121 gactgaggtg accgtccagc tcatcaatta caccaaatct ggcaaaaaat tctggaacct
     3181 gttccacctc cagcctatga gagatcagaa gggcgacgtc cagtacttta ttggcgtcca
     3241 gctggatgga actgaacatg tgagggacgc tgcagaacga gagggcgtga tgctcatcaa
     3301 aaaaaccgcc gaaaacatcg atgaggccgc taaagagctg tctagaagac ccaaacgacc
     3361 aaccacactg aacctcttct aggatccacc ggatctagat aactgatcat aatcagccat
     3421 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3481 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3541 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3601 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3661 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3721 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3781 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3841 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3901 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3961 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4021 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4081 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4141 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4201 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4261 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4321 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4381 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4441 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4501 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4561 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4621 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4681 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4741 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4801 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4861 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4921 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4981 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5041 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5101 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5161 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5221 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5281 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5341 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5401 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5461 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5521 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5581 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5641 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5701 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5761 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5821 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5881 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5941 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6001 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6061 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     6181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6721 cat
//
