LOCUS       sequence-315026-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optical regulation of nuclear export.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  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 5757)
  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-Dronpa-optoNES-WPRE
FEATURES             Location/Qualifiers
     source          1..5757
                     /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"
     CDS             694..1365
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     misc_feature    1886..2474
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1939..1959)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2357..2368)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2504..2980
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2703..2722)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3020..3160
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3020..3149
                     /label=AAV2 ITR
     rep_origin      3235..3690
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3322..3341)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3532..3553
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3707..3726)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3826..3848
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3886..3904)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3972..4076
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4077..4145,4146..4937)
                     /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(4295..4314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5108..5696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5597..5616
                     /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 accatgagtg tgattaaacc agacatgaag
      721 atcaagctgc gtatggaagg cgctgtaaat ggacacccgt tcgcgattga aggagttggc
      781 cttgggaagc ctttcgaggg aaaacagagt atggacctta aagtcaaaga aggcggacct
      841 ctgcctttcg cctatgacat cttgacaact gtgttctgtt acggcaacag ggtattcgcc
      901 aaatacccag aaaatatagt agactatttc aagcagtcgt ttcctgaggg ctactcttgg
      961 gaacgaagca tgaattacga agacgggggc atttgtaacg cgacaaacga cataaccctg
     1021 gatggtgact gttatatcta tgaaattcga tttgatggtg tgaactttcc tgccaatggt
     1081 ccagttatgc agaagaggac tgtgaaatgg gagccatcca ctgagaaatt gtatgtgcgt
     1141 gatggagtgc tgaagggtga tgttaacatg gctctgtcgc ttgaaggagg tggccattac
     1201 cgatgtgact tcaaaactac ttataaagct aagaaggttg tccagttgcc agactatcac
     1261 tttgtggacc accacattga gattaaaagc cacgacaaag attacagtaa tgttaatctg
     1321 catgagcatg ccgaagcgca ttctgagctg ccgaggcagg ccaagctgta caagtccgga
     1381 ctcagatctc gagctcaagc ttcgaattca ctggcaacaa cactggagag aatcgagaaa
     1441 aacttcgtca tcaccgaccc tagactgccc gataatccta tcatcttcgc ctccgactca
     1501 tttctccagc tcactgagta ctctcgggag gaaattctgg gccggaattg ccggttcctc
     1561 cagggacctg aaaccgatag agccactgtc cgaaaaatcc gggatgccat cgacaatcag
     1621 actgaggtga ccgtccagct catcaattac accaaatctg gcaaaaaatt ctggaacctg
     1681 ttccacctcc agcctatgag agatcagaag ggcgacgtcc agtactttat tggcgtccag
     1741 ctggatggaa ctgaacatgt gagggacgct gcagaacgag agggcgtgat gctcatcaaa
     1801 aaaaccgccg aaaacatcga tgagctgctg aaggagctgg ccgacctgaa cctggactag
     1861 ggatccgcgg ccgctagata actgaaatca acctctggat tacaaaattt gtgaaagatt
     1921 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1981 tttgtatcat gctattgctt cccgtatggc cttcattttc tcctccttgt ataaatcctg
     2041 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2101 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2161 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2221 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2281 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     2341 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2401 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2461 ggccgcctcc ccgcatcgat tgtaccgcgg gcccgggatc tacgggtggc atccctgtga
     2521 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     2581 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     2641 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     2701 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     2761 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     2821 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     2881 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     2941 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     3001 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3061 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3121 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3181 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3241 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3301 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3361 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3421 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3481 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3541 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3601 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3661 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3721 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3781 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3841 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3901 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3961 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4021 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4081 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4141 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4201 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4261 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4321 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4381 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4441 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4501 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4561 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4621 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4681 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4741 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4801 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4861 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4921 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4981 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5041 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5101 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5161 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5221 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5281 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5341 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5401 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5461 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5521 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5581 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5641 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5701 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
