LOCUS       sequence-236953-        6869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish Tol1 construct for UAS-mediated expression of the 
            optogenetic opsin GtACR2 fused to the red fluorescent protein 
            tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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:GtACR2-tdTomato
FEATURES             Location/Qualifiers
     source          1..6869
                     /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(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             join(5068..5136,5137..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
//
