LOCUS       sequence-236921-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish Tol1 construct for UAS-mediated expression of the 
            optogenetic opsin Chronos fused to the red fluorescent protein 
            tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  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:Chronos-tdTomato
FEATURES             Location/Qualifiers
     source          1..6971
                     /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"
     primer_bind     complement(4575..4592)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4575..4591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4584..4606)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4800..4819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4919..4941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4979..4997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5065..5169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5170..5238,5239..6030)
                     /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(5388..5407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6201..6789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6690..6709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6943..6960
                     /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 accatggaaa cagccgccac aatgacccac gcctttatct cagccgtgcc tagcgccgaa
     1621 gccacaatta gaggcctgct gagcgccgca gcagtggtga caccagcagc agacgctcac
     1681 ggagaaacct ctaacgccac aacagccgga gccgatcacg gttgcttccc ccacatcaac
     1741 cacggaaccg agctgcagca caagatcgca gtgggactcc agtggttcac cgtgatcgtg
     1801 gctatcgtgc agctcatctt ctacggttgg cacagcttca aggccacaac cggctgggag
     1861 gaggtctacg tctgcgtgat cgagctcgtc aagtgcttca tcgagctgtt ccacgaggtc
     1921 gacagcccag ccacagtgta ccagaccaac ggaggagccg tgatttggct gcggtacagc
     1981 atgtggctcc tgacttgccc cgtgatcctg atccacctga gcaacctgac cggactgcac
     2041 gaagagtaca gcaagcggac catgaccatc ctggtgaccg acatcggcaa catcgtgtgg
     2101 gggatcacag ccgcctttac aaagggcccc ctgaagatcc tgttcttcat gatcggcctg
     2161 ttctacggcg tgacttgctt cttccagatc gccaaggtgt atatcgagag ctaccacacc
     2221 ctgcccaaag gcgtctgccg gaagatttgc aagatcatgg cctacgtctt cttctgctct
     2281 tggctgatgt tccccgtgat gttcatcgcc ggacacgagg gactgggcct gatcacacct
     2341 tacaccagcg gaatcggcca cctgatcctg gatctgatca gcaagaacac ttggggcttc
     2401 ctgggccacc acctgagagt gaagatccac gagcacatcc tgatccacgg cgacatccgg
     2461 aagacaacca ccatcaacgt ggccggcgag aacatggaga tcgagacctt cgtcgacgag
     2521 gaggaggagg gaggagtggc ggcaccggta gtagcagtga gtaagggcga ggaagtgatc
     2581 aaagagttca tgcggtttaa ggtgagaatg gaaggaagca tgaacggcca cgagttcgaa
     2641 attgagggag aaggagaggg acggccctac gagggcaccc agacagccaa gctgaaagtg
     2701 acaaagggcg ggcctctgcc attcgcttgg gacatcctga gcccacagtt tatgtacggc
     2761 tccaaggcct atgtgaaaca tccagctgac attcccgatt ataagaaact gagcttcccc
     2821 gaggggttta agtgggaaag agtgatgaac ttcgaggacg gaggcctggt gactgtgacc
     2881 caggacagct ccctgcagga tgggaccctg atctacaagg tgaaaatgag agggacaaat
     2941 tttccccctg atggacctgt gatgcagaag aaaactatgg gatgggaggc ctccaccgaa
     3001 aggctgtatc cacgcgacgg ggtgctgaaa ggagaaatcc accaggctct gaagctgaaa
     3061 gatgggggac attacctggt ggagttcaag acaatctaca tggccaagaa acctgtgcag
     3121 ctgccaggct actattacgt ggacacaaaa ctggatatca cttcacacaa cgaggactac
     3181 actattgtgg agcagtatga acggagcgag gggagacacc atctgttcct gggccatggg
     3241 actggaagta ccggctcagg gtctagtgga accgcctcaa gcgaggataa caatatggct
     3301 gtgatcaaag agttcatgag gtttaaggtg cgcatggagg gcagcatgaa tgggcacgaa
     3361 tttgagattg aaggagaggg cgaagggagg ccttacgagg gcacacagac tgccaagctg
     3421 aaagtgacca agggaggacc actgcctttc gcttgggata tcctgtctcc tcagtttatg
     3481 tacggaagta aggcctatgt caagcatccc gctgacattc ctgattacaa gaaactgtct
     3541 ttcccagagg gctttaagtg ggagagagtg atgaattttg aagatggagg cctggtgacc
     3601 gtgacacagg actcctctct gcaggatggc actctgatct acaaagtcaa aatgcgcggc
     3661 accaattttc cacccgatgg gcccgtgatg cagaagaaaa caatggggtg ggaggccagc
     3721 actgaacggc tgtatcctag agacggagtg ctgaagggcg aaatccacca ggccctgaag
     3781 ctgaaagacg gcggccacta cctggtggag ttcaaaacca tctacatggc caagaaacca
     3841 gtgcagctgc ccggctatta ctatgtggac accaagctgg atatcacatc ccacaatgaa
     3901 gactacacca ttgtggaaca gtatgagagg tctgaaggac gccaccatct gtttctgtac
     3961 ggcatggatg agctgtataa gtaaacatgg agatctcgag actagtaatt aagtctcagc
     4021 caccgttaac tgaacatgtc aaaacctgtg gagactgttg agatttgatg ttctgaaaag
     4081 ataaagccta taaataaaat gttgcccaaa tttcctgcct gatgtttttc tttgtctttg
     4141 ctacatggct ttgctgctcg gatcggctca ctctgtgtat gccacgttca ctttgtactc
     4201 tccttctcac ggtaggttta ttatttttag atgtgcagtt agtttctgtg aaataacaca
     4261 ccacacactg atattgtctg tgcattgact tggtgagtgc acattgtttt tgatcttgac
     4321 atatttatat ttgattgatc aggtgaactg tgtgaatcta aagtgctcca tacagatgtt
     4381 ctgcattgaa aatattctca ttttattagt ggaagtgagt gtatgccagc ggccgcggta
     4441 ccctgcagtc tagacatttc agttgacgaa gacaaacaaa gttctgttgt gactatgggg
     4501 ggggggggcg cctggggatg gtctcgcccg gggagtaatt cagggtagaa ccgccactgc
     4561 ctttagcgac attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4621 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4681 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     4741 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     4801 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4861 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4921 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4981 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5041 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5101 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5161 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5221 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5281 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5341 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5401 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5461 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5521 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5581 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5641 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5701 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5761 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5821 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5881 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5941 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6001 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6061 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6121 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6181 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6241 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6301 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6361 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6421 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6481 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6541 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6601 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6661 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6721 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6781 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6841 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6901 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6961 gaagcggaag a
//
