LOCUS Exported 6944 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Zebrafish Tol1 construct for UAS-mediated expression of the optogenetic opsin CheRiff fused to the red fluorescent protein tdTomato. ACCESSION . VERSION . KEYWORDS pTol1-UAS:CheRiff-tdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6944) AUTHORS Antinucci P, Dumitrescu A, Deleuze C, Morley HJ, Leung K, Hagley T, Kubo F, Baier H, Bianco IH, Wyart C TITLE A calibrated optogenetic toolbox of stable zebrafish opsin lines. JOURNAL Elife. 2020 Mar 27;9. pii: 54937. doi: 10.7554/eLife.54937. PUBMED 32216873 REFERENCE 2 (bases 1 to 6944) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6944 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 106..127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 185..207 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 204..220 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" regulatory 1558..1567 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1564..2511 /codon_start=1 /product="blue light-activated optogenetic actuator derived from Scherffelia dubia channelrhodopsin (Hochbaum et al., 2014)" /label=CheRiff /translation="MGGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQ CSSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKV ILEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALL VSDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLV LAMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKI HEHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV" primer_bind complement(4548..4565) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4548..4564) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4557..4579) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4773..4792) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4892..4914 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4952..4970) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5038..5142 /gene="bla" /label=AmpR promoter CDS 5143..6003 /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(5361..5380) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6174..6762 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6663..6682 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6916..6933 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 241 atggtccctt tagccagtag cggttctagg cacgggccgt ccgggcggtg gcctggggcg 301 gaaaactgaa ggggggcggc accggcggct cagcccttgt aatatattat atgcaccact 361 attggtttac ttatgtcaca gtttgtaagt ttgtaacagc ctgaacctgg atcagcttag 421 gcctccaagg cggagtactg tcctccgggc tggcggagta ctgtcctccg gcaaggtcgg 481 agtactgtcc tccgacacta gaggtcggag tactgtcctc cgacgcaagg cggagtactg 541 tcctccgggc tgcggagtac tgtcctccgg caaggtcgga gtactgtcct ccgacactag 601 aggtcggagt actgtcctcc gacgcaaggt cggagtactg tcctccgaca ctagaggtcg 661 gagtactgtc ctccgacgca aggtcggagt actgtcctcc gacactagag gtcggagtac 721 tgtcctccga cgcaaggcgg agtactgtcc tccgggctgg cggagtactg tcctccggca 781 agggtcgact ctagagggta tataatggat cccatcgcgt ctcagcctca ctttgagctc 841 ctccacacga attcgatact aaggatcctt gtttgtgagt atgatgttaa atataagcaa 901 tcgacgtttg ttttgtcgtt tgttaaatct agcaatgttt tatttcaatt aaagtgttaa 961 ccggttcatt taaaggtgta tttctgaatg agctaacgta ttagctttta acgctagccg 1021 gactgtttgg ttttgaggat caataaacca acgttttttt ctgatttatt ttgctttaaa 1081 atataaaggt ataattcgtc gtcgagaatt ttgggttttg ttcgagttta gaaatttggc 1141 ggcgtttttt tgtaccgcga aacagggcgg tggttcggtg agcttctcaa ttcatctgct 1201 tgacaaacgg ttttcggcaa ctgtaaatgc gctatgatgc ttttattctt ttcagcgtgt 1261 aactttgtta aatataattg actttccgac tatcgacgtt cgtttcatct ccgtgttatt 1321 gtttaatggc cattttctgt ggtaaactgt cgccattttg ttttcagtcg tccgtttgtt 1381 ccgcccttac tcacagctgt gtcagcatta gtgatgactg ttctcacctg gcctgtttgc 1441 tttcattatt gatgaagagc tttctagaga attctgaagc tcgtctttgt cctgaattca 1501 tctgtattca tttaaaaaaa aaaattaacc ttaatttttt tttcttctca gcgtaaagcc 1561 accatgggcg gagctcctgc tccagacgct cacagcgccc cacctggaaa cgattctgcc 1621 ggaggcagtg agtaccatgc cccagctgga tatcaagtga atccacccta ccaccccgtg 1681 catgggtatg aggaacagtg cagctccatc tacatctact atggggccct gtgggagcag 1741 gaaacagcta ggggcttcca gtggtttgcc gtgttcctgt ctgccctgtt tctggctttc 1801 tacggctggc acgcctataa ggccagcgtg ggatgggagg aagtgtacgt gtgctccgtg 1861 gagctgatca aagtgattct ggagatctat ttcgagttca ccagtcctgc tatgctgttc 1921 ctgtacggag ggaacattac cccatggctg agatatgccg aatggctgct gacatgtccc 1981 gtgatcctga ttcatctgtc taacatcacc ggcctgagtg aggcatacaa taagcggaca 2041 atggctctgc tggtgtccga cctgggaact atttgcatgg gagtgacagc cgctctggcc 2101 actgggtggg tgaagtggct gttttactgt atcggcctgg tgtatggaac ccagacattc 2161 tacaacgctg gaatcatcta cgtggagtct tactatatca tgcctgccgg cggctgtaag 2221 aaactggtgc tggccatgac tgccgtgtac tattctagtt ggctgatgtt tcccggcctg 2281 ttcatctttg ggcctgaagg catgcacacc ctgagcgtgg ctgggtccac tattggccat 2341 accatcgccg acctgctgtc caagaatatt tggggactgc tggggcactt cctgcggatc 2401 aaaattcacg agcatatcat tatgtacggc gatatcagga gaccagtgag ctcccagttt 2461 ctgggacgca aggtggacgt gctggccttc gtgacagagg aagataaagt ggcggcaccg 2521 gtagtagcag tgagtaaggg cgaggaagtg atcaaagagt tcatgcggtt taaggtgaga 2581 atggaaggaa gcatgaacgg ccacgagttc gaaattgagg gagaaggaga gggacggccc 2641 tacgagggca cccagacagc caagctgaaa gtgacaaagg gcgggcctct gccattcgct 2701 tgggacatcc tgagcccaca gtttatgtac ggctccaagg cctatgtgaa acatccagct 2761 gacattcccg attataagaa actgagcttc cccgaggggt ttaagtggga aagagtgatg 2821 aacttcgagg acggaggcct ggtgactgtg acccaggaca gctccctgca ggatgggacc 2881 ctgatctaca aggtgaaaat gagagggaca aattttcccc ctgatggacc tgtgatgcag 2941 aagaaaacta tgggatggga ggcctccacc gaaaggctgt atccacgcga cggggtgctg 3001 aaaggagaaa tccaccaggc tctgaagctg aaagatgggg gacattacct ggtggagttc 3061 aagacaatct acatggccaa gaaacctgtg cagctgccag gctactatta cgtggacaca 3121 aaactggata tcacttcaca caacgaggac tacactattg tggagcagta tgaacggagc 3181 gaggggagac accatctgtt cctgggccat gggactggaa gtaccggctc agggtctagt 3241 ggaaccgcct caagcgagga taacaatatg gctgtgatca aagagttcat gaggtttaag 3301 gtgcgcatgg agggcagcat gaatgggcac gaatttgaga ttgaaggaga gggcgaaggg 3361 aggccttacg agggcacaca gactgccaag ctgaaagtga ccaagggagg accactgcct 3421 ttcgcttggg atatcctgtc tcctcagttt atgtacggaa gtaaggccta tgtcaagcat 3481 cccgctgaca ttcctgatta caagaaactg tctttcccag agggctttaa gtgggagaga 3541 gtgatgaatt ttgaagatgg aggcctggtg accgtgacac aggactcctc tctgcaggat 3601 ggcactctga tctacaaagt caaaatgcgc ggcaccaatt ttccacccga tgggcccgtg 3661 atgcagaaga aaacaatggg gtgggaggcc agcactgaac ggctgtatcc tagagacgga 3721 gtgctgaagg gcgaaatcca ccaggccctg aagctgaaag acggcggcca ctacctggtg 3781 gagttcaaaa ccatctacat ggccaagaaa ccagtgcagc tgcccggcta ttactatgtg 3841 gacaccaagc tggatatcac atcccacaat gaagactaca ccattgtgga acagtatgag 3901 aggtctgaag gacgccacca tctgtttctg tacggcatgg atgagctgta taagtaaaca 3961 tggagatctc gagactagta attaagtctc agccaccgtt aactgaacat gtcaaaacct 4021 gtggagactg ttgagatttg atgttctgaa aagataaagc ctataaataa aatgttgccc 4081 aaatttcctg cctgatgttt ttctttgtct ttgctacatg gctttgctgc tcggatcggc 4141 tcactctgtg tatgccacgt tcactttgta ctctccttct cacggtaggt ttattatttt 4201 tagatgtgca gttagtttct gtgaaataac acaccacaca ctgatattgt ctgtgcattg 4261 acttggtgag tgcacattgt ttttgatctt gacatattta tatttgattg atcaggtgaa 4321 ctgtgtgaat ctaaagtgct ccatacagat gttctgcatt gaaaatattc tcattttatt 4381 agtggaagtg agtgtatgcc agcggccgcg gtaccctgca gtctagacat ttcagttgac 4441 gaagacaaac aaagttctgt tgtgactatg gggggggggg gcgcctgggg atggtctcgc 4501 ccggggagta attcagggta gaaccgccac tgcctttagc gacattcact ggccgtcgtt 4561 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 4621 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 4681 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc 4741 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 4801 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 4861 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 4921 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 4981 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 5041 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 5101 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 5161 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 5221 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 5281 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 5341 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 5401 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 5461 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 5521 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 5581 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 5641 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 5701 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 5761 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 5821 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 5881 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 5941 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 6001 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 6061 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 6121 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 6181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 6241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 6301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 6361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 6421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 6481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 6541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 6601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 6661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 6721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 6781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 6841 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 6901 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga //