LOCUS       sequence-315045-        5772 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 5772)
  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 5772)
  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-mScarlet-optoNES-WPRE
FEATURES             Location/Qualifiers
     source          1..5772
                     /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             694..1389
                     /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(827..845)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1095..1114
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1901..2489
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1954..1974)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2372..2383)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2519..2995
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2718..2737)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3035..3175
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3035..3164
                     /label=AAV2 ITR
     rep_origin      3250..3705
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3337..3356)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3547..3568
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3722..3741)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3841..3863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3901..3919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3987..4091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4092..4160,4161..4952)
                     /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(4310..4329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5123..5711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5612..5631
                     /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 ggcagtgatc
      721 aaggagttca tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag
      781 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      841 accaagggtg gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc
      901 tccagggcct tcaccaagca ccccgccgac atccccgact actataagca gtccttcccc
      961 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc
     1021 caggacacct ccctggagga cggcaccctg atctacaagg tgaagctccg cggcaccaac
     1081 ttccctcctg acggccccgt aatgcagaag aagacaatgg gctgggaagc gtccaccgag
     1141 cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta agatggccct gcgcctgaag
     1201 gacggcggcc gctacctggc ggacttcaag accacctaca aggccaagaa gcccgtgcag
     1261 atgcccggcg cctacaacgt cgaccgcaag ttggacatca cctcccacaa cgaggactac
     1321 accgtggtgg aacagtacga acgctccgag ggccgccact ccaccggcgg catggacgag
     1381 ctgtacaagt ccggactcag atctcgagct caagcttcga attcactggc aacaacactg
     1441 gagagaatcg agaaaaactt cgtcatcacc gaccctagac tgcccgataa tcctatcatc
     1501 ttcgcctccg actcatttct ccagctcact gagtactctc gggaggaaat tctgggccgg
     1561 aattgccggt tcctccaggg acctgaaacc gatagagcca ctgtccgaaa aatccgggat
     1621 gccatcgaca atcagactga ggtgaccgtc cagctcatca attacaccaa atctggcaaa
     1681 aaattctgga acctgttcca cctccagcct atgagagatc agaagggcga cgtccagtac
     1741 tttattggcg tccagctgga tggaactgaa catgtgaggg acgctgcaga acgagagggc
     1801 gtgatgctca tcaaaaaaac cgccgaaaac atcgatgagc tgctgaagga gctggccgac
     1861 ctgaacctgg actagggatc cgcggccgct agataactga aatcaacctc tggattacaa
     1921 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1981 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggccttca ttttctcctc
     2041 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2101 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2161 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2221 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2281 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     2341 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2401 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2461 tcggatctcc ctttgggccg cctccccgca tcgattgtac cgcgggcccg ggatctacgg
     2521 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2581 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2641 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2701 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2761 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2821 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2881 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     2941 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3001 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3061 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3121 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3241 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3301 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3361 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3421 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3481 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3541 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     3601 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     3661 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3721 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3781 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3841 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3901 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4381 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4441 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4501 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4561 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4621 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4681 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4741 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4801 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4861 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4921 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4981 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5041 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5101 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5161 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5221 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5281 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5341 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5401 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5461 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5521 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5581 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5641 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5701 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5761 ttgctcacat gt
//
