LOCUS Exported 5403 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION optical regulation of nuclear export. ACCESSION . VERSION . KEYWORDS AAV-Synapsin-mTurquoise-3XNLS-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5403) 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 5403) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5403 /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 652..661 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 658..1374 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(703..724) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(964..983) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1311..1332 /label=EGFP-C /note="EGFP, forward primer" CDS 1417..1437 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 1456..1512 /label=MCS /note="multiple cloning site" misc_feature 1532..2120 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1585..1605) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2003..2014) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2150..2626 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2349..2368) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2666..2806 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2666..2795 /label=AAV2 ITR rep_origin 2881..3336 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2968..2987) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3178..3199 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3353..3372) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3472..3494 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3532..3550) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3618..3722 /gene="bla" /label=AmpR promoter CDS 3723..4583 /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(3941..3960) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4754..5342 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5243..5262 /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 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 721 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 781 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 841 gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag 901 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 961 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 1021 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 1081 ctggagtaca actactttag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc 1141 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac 1201 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 1261 ctgagcaccc agtccaagct tagcaaagac cccaacgaga agcgcgatca catggtcctg 1321 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga 1381 ctcagatccg accccagaag aaagcgcaag gtggacccca agaagaagcg caaggtggac 1441 cccaagaaga agcgcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg 1501 ggcccgggat ccgcggccgc tagataactg aaatcaacct ctggattaca aaatttgtga 1561 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 1621 aatgcctttg tatcatgcta ttgcttcccg tatggccttc attttctcct ccttgtataa 1681 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 1741 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 1801 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 1861 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 1921 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg 1981 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 2041 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 2101 cctttgggcc gcctccccgc atcgattgta ccgcgggccc gggatctacg ggtggcatcc 2161 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 2221 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 2281 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 2341 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 2401 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 2461 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 2521 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 2581 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 2641 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 2701 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 2761 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 2821 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 2881 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 2941 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3001 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 3061 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 3121 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3181 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 3241 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 3301 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 3361 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 3421 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 3481 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 3541 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 3601 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 3661 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 3721 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 3781 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 3841 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 3901 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 3961 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 4021 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 4081 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 4141 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 4201 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 4261 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 4321 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 4381 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 4441 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 4501 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 4561 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 4621 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 4681 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 4741 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 4801 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 4861 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 4921 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 4981 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 5041 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 5101 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 5161 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 5221 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 5281 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 5341 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 5401 tgt //