LOCUS       Exported                5784 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  optical regulation of nuclear export with shorter relaxation time.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-Synapsin-NLS-YPET-optoNES.F434L-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  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 5784)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5784
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             688..1401
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELYK"
     primer_bind     complement(730..751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(991..1010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1913..2501
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1966..1986)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2384..2395)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2531..3007
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2730..2749)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3047..3187
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3047..3176
                     /label=AAV2 ITR
     rep_origin      3262..3717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3349..3368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3559..3580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3734..3753)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3853..3875
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3913..3931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3999..4103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4104..4964
                     /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(4322..4341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5135..5723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5624..5643
                     /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 agctagacct accggtcgtg agcaagggcg aggagctgtt caccggggtg
      721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agcttctatg caccaccggc
      841 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
      901 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1081 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1141 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1201 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1261 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgtt caaagacccc
     1321 aacgagaagc gcgatcacat ggtcctgctg gagttcctga ccgccgccgg gatcactgag
     1381 ggcatgaacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattcactg
     1441 gcaacaacac tggagagaat cgagaaaaac ttcgtcatca ccgaccctag actgcccgat
     1501 aatcctatca tcttcgcctc cgactcactc ctccagctca ctgagtactc tcgggaggaa
     1561 attctgggcc ggaattgccg gttcctccag ggacctgaaa ccgatagagc cactgtccga
     1621 aaaatccggg atgccatcga caatcagact gaggtgaccg tccagctcat caattacacc
     1681 aaatctggca aaaaattctg gaacctgttc cacctccagc ctatgagaga tcagaagggc
     1741 gacgtccagt actttattgg cgtccagctg gatggaactg aacatgtgag ggacgctgca
     1801 gaacgagagg gcgtgatgct catcaaaaaa accgccgaaa acatcgatga gctgctgaag
     1861 gagctggccg acctgaacct ggactaggga tccgcggccg ctagataact gaaatcaacc
     1921 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1981 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcctt
     2041 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2101 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2161 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2221 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2281 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     2341 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2401 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2461 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgattgt accgcgggcc
     2521 cgggatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     2581 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2641 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2701 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     2761 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     2821 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     2881 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     2941 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3001 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     3061 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3121 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3181 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3241 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3301 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     3361 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3421 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3481 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3541 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3601 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3661 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3721 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3781 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3841 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3901 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3961 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4021 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4081 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4141 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4201 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4261 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4321 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4381 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4441 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4501 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4561 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4621 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4681 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4741 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4801 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4861 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4921 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5761 tttgctggcc ttttgctcac atgt
//
