LOCUS       sequence-315114-        5808 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 5808)
  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 5808)
  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-3XNLS-optoNES-WPRE
FEATURES             Location/Qualifiers
     source          1..5808
                     /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"
     CDS             1396..1416
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1937..2525
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1990..2010)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2408..2419)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2555..3031
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2754..2773)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3071..3211
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3071..3200
                     /label=AAV2 ITR
     rep_origin      3286..3741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3373..3392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3583..3604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3758..3777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3877..3899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3937..3955)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4023..4127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4128..4196,4197..4988)
                     /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(4346..4365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5159..5747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5648..5667
                     /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 tcagatccga ccccagaaga
     1381 aagcgcaagg tggaccccaa gaagaagcgc aaggtggacc ccaagaagaa gcgcagatct
     1441 cgagctcaag cttcgaattc actggcaaca acactggaga gaatcgagaa aaacttcgtc
     1501 atcaccgacc ctagactgcc cgataatcct atcatcttcg cctccgactc atttctccag
     1561 ctcactgagt actctcggga ggaaattctg ggccggaatt gccggttcct ccagggacct
     1621 gaaaccgata gagccactgt ccgaaaaatc cgggatgcca tcgacaatca gactgaggtg
     1681 accgtccagc tcatcaatta caccaaatct ggcaaaaaat tctggaacct gttccacctc
     1741 cagcctatga gagatcagaa gggcgacgtc cagtacttta ttggcgtcca gctggatgga
     1801 actgaacatg tgagggacgc tgcagaacga gagggcgtga tgctcatcaa aaaaaccgcc
     1861 gaaaacatcg atgagctgct gaaggagctg gccgacctga acctggacta gggatccgcg
     1921 gccgctagat aactgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1981 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2041 tgctattgct tcccgtatgg ccttcatttt ctcctccttg tataaatcct ggttgctgtc
     2101 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2161 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2221 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2281 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2341 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     2401 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2461 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2521 cccgcatcga ttgtaccgcg ggcccgggat ctacgggtgg catccctgtg acccctcccc
     2581 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     2641 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     2701 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     2761 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     2821 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     2881 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     2941 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     3001 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3061 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3121 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3181 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3241 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3301 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3361 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3421 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3481 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3541 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3601 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3661 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3721 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3781 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3841 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3901 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3961 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
