LOCUS Exported 5814 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION optoNES construct with modified mTurquoise2-LOV2 RET. ACCESSION . VERSION . KEYWORDS AAV-CMV-NLS-mTurquoise2dC10-408LOV2-Ja(delta)-NES21-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5814) 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 5814) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5814 /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 1943..2531 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1996..2016) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2414..2425) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2561..3037 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2760..2779) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3077..3217 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3077..3206 /label=AAV2 ITR rep_origin 3292..3747 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3379..3398) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3589..3610 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3764..3783) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3883..3905 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3943..3961) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4029..4133 /gene="bla" /label=AmpR promoter CDS 4134..4994 /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(4352..4371) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5165..5753 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5654..5673 /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 gccgccgggc tcgagagaat cgagaaaaac 1501 ttcgtcatca ccgaccctag actgcccgat aatcctatca tcttcgcctc cgactcattt 1561 ctccagctca ctgagtactc tcgggaggaa attctgggcc ggaattgccg gttcctccag 1621 ggacctgaaa ccgatagagc cactgtccga aaaatccggg atgccatcga caatcagact 1681 gaggtgaccg tccagctcat caattacacc aaatctggca aaaaattctg gaacctgttc 1741 cacctccagc ctatgagaga tcagaagggc gacgtccagt actttattgg cgtccagctg 1801 gatggaactg aacatgtgag ggacgctgca gaacgagagg gcgtgatgct catcaaaaaa 1861 accgccgaaa acatcgatga gctgctgaag gagctggccg acctgaacct ggactaggga 1921 tccgcggccg ctagataact gaaatcaacc tctggattac aaaatttgtg aaagattgac 1981 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 2041 gtatcatgct attgcttccc gtatggcctt cattttctcc tccttgtata aatcctggtt 2101 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 2161 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 2221 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 2281 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 2341 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt 2401 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 2461 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 2521 cgcctccccg catcgattgt accgcgggcc cgggatctac gggtggcatc cctgtgaccc 2581 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct 2641 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg 2701 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt 2761 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc 2821 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat 2881 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag 2941 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg 3001 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg 3061 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct 3121 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg 3181 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc 3241 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct 3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 3361 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 3481 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct 3541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 3601 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt 3661 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 3721 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg 3781 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 3841 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 3901 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 3961 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 4021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 4081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 4141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 4201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 4261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 4321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 4381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 4441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 4501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 4561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 4621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 4681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 4741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 4801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 4861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 4921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 4981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 5041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 5101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 5161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 5221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 5281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 5341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 5401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 5461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 5521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 5581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 5641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 5701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 5761 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt //