LOCUS Exported 6869 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Zebrafish Tol1 construct for UAS-mediated expression of the optogenetic opsin GtACR2 fused to the red fluorescent protein tdTomato. ACCESSION . VERSION . KEYWORDS pTol1-UAS:GtACR2-tdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6869) 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 6869) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6869 /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)" primer_bind complement(4473..4490) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4473..4489) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4482..4504) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4698..4717) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4817..4839 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4877..4895) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4963..5067 /gene="bla" /label=AmpR promoter CDS 5068..5928 /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(5286..5305) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6099..6687 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6588..6607 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6841..6858 /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 accatggcct cccaggtcgt gtacggcgag tgggccagca cccacacaga gtgctacaac 1621 atgagcagga tcgacagcac cttcgtgagc ctgctgcagc tggtgtgggc cgtggtgagc 1681 ggatgccaga ccatcttcat gatcagcagg gcccccaagg tgccctggga gagcgtgtac 1741 ctgcccttcg tggagtccat cacatacgcc ctggccagca ccggcaatgg cacactgcag 1801 atgagggacg gcaggttctt tccctggtcc aggatggcca gctggctgtg cacatgcccc 1861 attatgctgg gccagatctc caacatggct ctggtgaagt acaagagcat ccctctgaac 1921 cccatcgccc aggccgccag cattatcagg gtggtgatgg gcattaccgc taccatctcc 1981 cccgccgagt acatgaagtg gctgttcttc ttcttcggcg ctacctgcct ggtcttcgaa 2041 tacagcgtgg tgttcaccat cttccaggtg ggactgtacg gcttcgagtc cgtgggcacc 2101 cctctggccc agaaggtggt ggtgaggatc aagatgctga ggctgatctt cttcatcgcc 2161 tggaccatgt tccccatcgt gtggctgatt tcccccaccg gcgtgtgcgt gatccacgag 2221 aacgtgagcg ccatcctgta tctgctcgcc gacggcctgt gcaagaacac ctacggcgtg 2281 atcctgtggt ccaccgcctg gggagtgctc gagggaaagt gggacccggc ttgcctccct 2341 ggccaggaga agcctgaggc cgacgacccc ttcggactga accacgagaa gaatgccccc 2401 cccaacgacg aggtcaacat cagaatgttc ggcagggcgg caccggtagt agcagtgagt 2461 aagggcgagg aagtgatcaa agagttcatg cggtttaagg tgagaatgga aggaagcatg 2521 aacggccacg agttcgaaat tgagggagaa ggagagggac ggccctacga gggcacccag 2581 acagccaagc tgaaagtgac aaagggcggg cctctgccat tcgcttggga catcctgagc 2641 ccacagttta tgtacggctc caaggcctat gtgaaacatc cagctgacat tcccgattat 2701 aagaaactga gcttccccga ggggtttaag tgggaaagag tgatgaactt cgaggacgga 2761 ggcctggtga ctgtgaccca ggacagctcc ctgcaggatg ggaccctgat ctacaaggtg 2821 aaaatgagag ggacaaattt tccccctgat ggacctgtga tgcagaagaa aactatggga 2881 tgggaggcct ccaccgaaag gctgtatcca cgcgacgggg tgctgaaagg agaaatccac 2941 caggctctga agctgaaaga tgggggacat tacctggtgg agttcaagac aatctacatg 3001 gccaagaaac ctgtgcagct gccaggctac tattacgtgg acacaaaact ggatatcact 3061 tcacacaacg aggactacac tattgtggag cagtatgaac ggagcgaggg gagacaccat 3121 ctgttcctgg gccatgggac tggaagtacc ggctcagggt ctagtggaac cgcctcaagc 3181 gaggataaca atatggctgt gatcaaagag ttcatgaggt ttaaggtgcg catggagggc 3241 agcatgaatg ggcacgaatt tgagattgaa ggagagggcg aagggaggcc ttacgagggc 3301 acacagactg ccaagctgaa agtgaccaag ggaggaccac tgcctttcgc ttgggatatc 3361 ctgtctcctc agtttatgta cggaagtaag gcctatgtca agcatcccgc tgacattcct 3421 gattacaaga aactgtcttt cccagagggc tttaagtggg agagagtgat gaattttgaa 3481 gatggaggcc tggtgaccgt gacacaggac tcctctctgc aggatggcac tctgatctac 3541 aaagtcaaaa tgcgcggcac caattttcca cccgatgggc ccgtgatgca gaagaaaaca 3601 atggggtggg aggccagcac tgaacggctg tatcctagag acggagtgct gaagggcgaa 3661 atccaccagg ccctgaagct gaaagacggc ggccactacc tggtggagtt caaaaccatc 3721 tacatggcca agaaaccagt gcagctgccc ggctattact atgtggacac caagctggat 3781 atcacatccc acaatgaaga ctacaccatt gtggaacagt atgagaggtc tgaaggacgc 3841 caccatctgt ttctgtacgg catggatgag ctgtataagt aaacatggag atctcgagac 3901 tagtaattaa gtctcagcca ccgttaactg aacatgtcaa aacctgtgga gactgttgag 3961 atttgatgtt ctgaaaagat aaagcctata aataaaatgt tgcccaaatt tcctgcctga 4021 tgtttttctt tgtctttgct acatggcttt gctgctcgga tcggctcact ctgtgtatgc 4081 cacgttcact ttgtactctc cttctcacgg taggtttatt atttttagat gtgcagttag 4141 tttctgtgaa ataacacacc acacactgat attgtctgtg cattgacttg gtgagtgcac 4201 attgtttttg atcttgacat atttatattt gattgatcag gtgaactgtg tgaatctaaa 4261 gtgctccata cagatgttct gcattgaaaa tattctcatt ttattagtgg aagtgagtgt 4321 atgccagcgg ccgcggtacc ctgcagtcta gacatttcag ttgacgaaga caaacaaagt 4381 tctgttgtga ctatgggggg ggggggcgcc tggggatggt ctcgcccggg gagtaattca 4441 gggtagaacc gccactgcct ttagcgacat tcactggccg tcgttttaca acgtcgtgac 4501 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 4561 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 4621 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 4681 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac 4741 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga 4801 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa 4861 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata 4921 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 4981 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 5041 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt 5101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 5161 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc 5221 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa 5281 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 5341 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 5401 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact 5461 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac 5521 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 5581 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta 5641 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg 5701 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 5761 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt 5821 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga 5881 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa 5941 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 6001 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 6061 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 6121 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 6181 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 6241 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 6301 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 6361 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 6421 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 6481 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 6541 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 6601 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 6661 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 6721 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 6781 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 6841 agcgagtcag tgagcgagga agcggaaga //