LOCUS Exported 5823 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION optoNES construct with modified mTurquoise2-LOV2 RET. ACCESSION . VERSION . KEYWORDS AAV-CMV-NLS-mTurquoise2dC7-408LOV2-Ja(delta)-NES21-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5823) 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 5823) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5823 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" enhancer 205..508 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 509..712 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 663..683 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 742..751 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 787..796 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(838..859) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1099..1118) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1446..1467 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1952..2540 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2005..2025) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2423..2434) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2570..3046 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2769..2788) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3086..3226 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3086..3215 /label=AAV2 ITR rep_origin 3301..3756 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3388..3407) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3598..3619 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3773..3792) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3892..3914 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3952..3970) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4038..4142 /gene="bla" /label=AmpR promoter CDS 4143..5003 /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(4361..4380) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5174..5762 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5663..5682 /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 agaatagtaa tcaattacgg ggtcattagt 181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 481 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 541 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 601 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 661 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt 721 gaaccgtcag atccgctagc cgccaccatg gccgcagcaa agcgcgtgaa gctggaccta 781 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 841 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 901 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 961 ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc 1021 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 1081 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 1141 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1201 atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac 1261 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc 1321 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1381 cccgacaacc actacctgag cacccagtcc aagcttagca aagaccccaa cgagaagcgc 1441 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcct cgagagaatc 1501 gagaaaaact tcgtcatcac cgaccctaga ctgcccgata atcctatcat cttcgcctcc 1561 gactcatttc tccagctcac tgagtactct cgggaggaaa ttctgggccg gaattgccgg 1621 ttcctccagg gacctgaaac cgatagagcc actgtccgaa aaatccggga tgccatcgac 1681 aatcagactg aggtgaccgt ccagctcatc aattacacca aatctggcaa aaaattctgg 1741 aacctgttcc acctccagcc tatgagagat cagaagggcg acgtccagta ctttattggc 1801 gtccagctgg atggaactga acatgtgagg gacgctgcag aacgagaggg cgtgatgctc 1861 atcaaaaaaa ccgccgaaaa catcgatgag ctgctgaagg agctggccga cctgaacctg 1921 gactagggat ccgcggccgc tagataactg aaatcaacct ctggattaca aaatttgtga 1981 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 2041 aatgcctttg tatcatgcta ttgcttcccg tatggccttc attttctcct ccttgtataa 2101 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 2161 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 2221 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 2281 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 2341 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg 2401 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 2461 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 2521 cctttgggcc gcctccccgc atcgattgta ccgcgggccc gggatctacg ggtggcatcc 2581 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 2641 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 2701 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 2761 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 2821 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 2881 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 2941 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 3001 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 3061 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 3121 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 3181 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 3241 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3361 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 3481 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 3541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3601 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 3661 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 3721 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 3781 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 3841 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 3901 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 3961 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 4021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 4081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 4141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 4201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 4261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 4321 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 4381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 4441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 4501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 4561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 4621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 4681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 4741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 4801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 4861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 4921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 4981 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 5041 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 5101 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 5161 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 5221 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 5281 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 5341 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 5401 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 5461 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 5521 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 5581 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 5641 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 5701 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 5761 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 5821 tgt //