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.I427V-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 aatcctgtca tcttcgcctc cgactcattt 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 //