LOCUS       sequence-315098-        5715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optoNES construct with modified mTurquoise2-LOV2 RET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5715)
  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 5715)
  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-NLS-mTurquoise2dC10-408LOV2-Ja(delta)-NES21-WPRE
FEATURES             Location/Qualifiers
     source          1..5715
                     /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      643..652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(694..696,697..699,700..1212)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(739..760)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1000..1019)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1347..1368
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1844..2432
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1897..1917)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2315..2326)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2462..2938
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2661..2680)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2978..3118
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2978..3107
                     /label=AAV2 ITR
     rep_origin      3193..3648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3280..3299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3490..3511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3665..3684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3784..3806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3844..3862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3930..4034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4035..4103,4104..4895)
                     /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(4253..4272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5066..5654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5555..5574
                     /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 ccgccaccat ggccgcagca
      661 aagcgcgtga agctggacct accggtcgcc accatggtga gcaagggcga ggagctgttc
      721 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      781 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      841 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgtc ctggggcgtg
      901 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      961 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1021 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1081 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta ctttagcgac
     1141 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1201 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1261 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagcttagc
     1321 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1381 ctcgagagaa tcgagaaaaa cttcgtcatc accgacccta gactgcccga taatcctatc
     1441 atcttcgcct ccgactcatt tctccagctc actgagtact ctcgggagga aattctgggc
     1501 cggaattgcc ggttcctcca gggacctgaa accgatagag ccactgtccg aaaaatccgg
     1561 gatgccatcg acaatcagac tgaggtgacc gtccagctca tcaattacac caaatctggc
     1621 aaaaaattct ggaacctgtt ccacctccag cctatgagag atcagaaggg cgacgtccag
     1681 tactttattg gcgtccagct ggatggaact gaacatgtga gggacgctgc agaacgagag
     1741 ggcgtgatgc tcatcaaaaa aaccgccgaa aacatcgatg agctgctgaa ggagctggcc
     1801 gacctgaacc tggactaggg atccgcggcc gctagataac tgaaatcaac ctctggatta
     1861 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1921 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggcct tcattttctc
     1981 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2041 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2101 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2161 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2221 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     2281 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2341 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2401 gagtcggatc tccctttggg ccgcctcccc gcatcgattg taccgcgggc ccgggatcta
     2461 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     2521 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     2581 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     2641 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     2701 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     2761 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     2821 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     2881 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     2941 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3001 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3061 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3121 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3181 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3241 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     3301 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3361 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3421 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3481 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3541 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3601 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3661 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3721 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3781 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3841 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3901 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3961 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4021 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4081 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4141 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4201 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4261 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4321 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4381 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4441 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4501 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4561 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4621 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4681 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4741 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4801 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4861 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4921 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4981 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5041 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5641 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5701 cttttgctca catgt
//
