LOCUS       sequence-236930-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish Tol1 construct for UAS-mediated expression of the 
            optogenetic opsin CheRiff fused to the red fluorescent protein 
            tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2020-03-27"; volume: "9. pii: 54937. doi"; pages: " 
            10.7554/eLife.54937"
COMMENT     Sequence Label: pTol1-UAS:CheRiff-tdTomato
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        join(142..147,148..165,166..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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             join(5143..5211,5212..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
//
