LOCUS       sequence-315051-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optogenetic JNK inhibitor with increased RET from LOV2 to reduce 
            actuation sensitivity, detectable without activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  AUTHORS   Li LL, Klein FM, Li Greci L, Popinigis A, Freudenberg F, Courtney MJ
  TITLE     Resonance energy transfer sensitises and monitors in situ switching 
            of LOV2-based optogenetic actuators.
  JOURNAL   Nat Commun. 2020 Oct 9;11(1):5107. doi: 10.1038/s41467-020-18816-8.
  PUBMED    33037199
REFERENCE   2  (bases 1 to 5756)
  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: "2020-10-9"; volume: "11(1):5107. doi"; pages: " 
            10.1038/s41467-020-18816-8"
COMMENT     Sequence Label: AAV-Synapsin-mScarlet-optoJNKi-WPRE
FEATURES             Location/Qualifiers
     source          1..5756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             658..1353
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(791..809)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1059..1078
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1885..2473
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1938..1958)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2356..2367)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2503..2979
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2702..2721)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3019..3159
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3019..3148
                     /label=AAV2 ITR
     rep_origin      3234..3689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3321..3340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3531..3552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3706..3725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3825..3847
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3885..3903)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3971..4075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4076..4144,4145..4936)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4294..4313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5107..5695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5596..5615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtacagctag cgctaccggt cgccaccatg
      661 gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc
      721 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      781 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc
      841 ctgtcccctc agttcatgta cggctccagg gccttcacca agcaccccgc cgacatcccc
      901 gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      961 gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac
     1021 aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca
     1081 atgggctggg aagcgtccac cgagcggttg taccccgagg acggcgtgct gaagggcgac
     1141 attaagatgg ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc
     1201 tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac
     1261 atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc
     1321 cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctcg agctcaagct
     1381 tcgaattcac tggcaacaac actggagaga atcgagaaaa acttcgtcat caccgaccct
     1441 agactgcccg ataatcctat catcttcgcc tccgactcat ttctccagct cactgagtac
     1501 tctcgggagg aaattctggg ccggaattgc cggttcctcc agggacctga aaccgataga
     1561 gccactgtcc gaaaaatccg ggatgccatc gacaatcaga ctgaggtgac cgtccagctc
     1621 atcaattaca ccaaatctgg caaaaaattc tggaacctgt tccacctcca gcctatgaga
     1681 gatcagaagg gcgacgtcca gtactttatt ggcgtccagc tggatggaac tgaacatgtg
     1741 agggacgctg cagaacgaga gggcgtgatg ctcatcaaaa aaaccgccga aaacatcgat
     1801 gaggccgcta aagagctgtc tagaagaccc aaacgaccaa ccacactgaa cctcttctag
     1861 gatccgcggc cgctagataa ctgaaatcaa cctctggatt acaaaatttg tgaaagattg
     1921 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1981 ttgtatcatg ctattgcttc ccgtatggcc ttcattttct cctccttgta taaatcctgg
     2041 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2101 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2161 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2221 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2281 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2341 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2401 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2461 gccgcctccc cgcatcgatt gtaccgcggg cccgggatct acgggtggca tccctgtgac
     2521 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2581 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2641 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2701 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2761 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2821 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2881 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2941 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3001 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3061 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3121 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3181 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3241 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3301 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3361 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3421 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3481 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3541 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3601 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3661 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3721 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3781 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3841 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3901 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3961 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4021 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4081 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4141 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4201 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4261 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4321 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4381 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4441 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4501 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4561 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4621 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4681 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4741 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4801 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4861 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4921 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5221 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
