LOCUS       sequence-236945-        6923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish Tol1 construct for UAS-mediated expression of the 
            optogenetic opsin ChR2(H134R) fused to the red fluorescent protein 
            tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6923)
  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 6923)
  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:ChR2(H134R)-tdTomato
FEATURES             Location/Qualifiers
     source          1..6923
                     /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..2490
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     primer_bind     complement(4527..4544)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4527..4543)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4536..4558)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4752..4771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4871..4893
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4931..4949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5017..5121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5122..5190,5191..5982)
                     /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(5340..5359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6153..6741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6642..6661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6895..6912
                     /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 accatggact atggcggcgc tttgtctgcc gtcggacgcg aacttttgtt cgttactaat
     1621 cctgtggtgg tgaacgggtc cgtcctggtc cctgaggatc aatgttactg tgccggatgg
     1681 attgaatctc gcggcacgaa cggcgctcag accgcgtcaa atgtcctgca gtggcttgca
     1741 gcaggattca gcattttgct gctgatgttc tatgcctacc aaacctggaa atctacatgc
     1801 ggctgggagg agatctatgt gtgcgccatt gaaatggtta aggtgattct cgagttcttt
     1861 tttgagttta agaatccctc tatgctctac cttgccacag gacaccgggt gcagtggctg
     1921 cgctatgcag agtggctgct cacttgtcct gtcatcctta tcagactgag caacctcacc
     1981 ggcctgagca acgactacag caggagaacc atgggactcc ttgtctcaga catcgggact
     2041 atcgtgtggg gggctaccag cgccatggca accggctatg ttaaagtcat cttcttttgt
     2101 cttggattgt gctatggcgc gaacacattt tttcacgccg ccaaagcata tatcgagggt
     2161 tatcatactg tgccaaaggg tcggtgccgc caggtcgtga ccggcatggc atggctgttt
     2221 ttcgtgagct ggggtatgtt cccaattctc ttcattttgg ggcccgaagg ttttggcgtc
     2281 ctgagcgtct atggctccac cgtaggtcac acgattattg atctgatgag taaaaattgt
     2341 tgggggttgt tgggacacta cctgcgcgtc ctgatccacg agcacatatt gattcacgga
     2401 gatatccgca aaaccaccaa actgaacatc ggcggaacgg agatcgaggt cgagactctc
     2461 gtcgaagacg aagccgaggc cggagccgtg gcggcaccgg tagtagcagt gagtaagggc
     2521 gaggaagtga tcaaagagtt catgcggttt aaggtgagaa tggaaggaag catgaacggc
     2581 cacgagttcg aaattgaggg agaaggagag ggacggccct acgagggcac ccagacagcc
     2641 aagctgaaag tgacaaaggg cgggcctctg ccattcgctt gggacatcct gagcccacag
     2701 tttatgtacg gctccaaggc ctatgtgaaa catccagctg acattcccga ttataagaaa
     2761 ctgagcttcc ccgaggggtt taagtgggaa agagtgatga acttcgagga cggaggcctg
     2821 gtgactgtga cccaggacag ctccctgcag gatgggaccc tgatctacaa ggtgaaaatg
     2881 agagggacaa attttccccc tgatggacct gtgatgcaga agaaaactat gggatgggag
     2941 gcctccaccg aaaggctgta tccacgcgac ggggtgctga aaggagaaat ccaccaggct
     3001 ctgaagctga aagatggggg acattacctg gtggagttca agacaatcta catggccaag
     3061 aaacctgtgc agctgccagg ctactattac gtggacacaa aactggatat cacttcacac
     3121 aacgaggact acactattgt ggagcagtat gaacggagcg aggggagaca ccatctgttc
     3181 ctgggccatg ggactggaag taccggctca gggtctagtg gaaccgcctc aagcgaggat
     3241 aacaatatgg ctgtgatcaa agagttcatg aggtttaagg tgcgcatgga gggcagcatg
     3301 aatgggcacg aatttgagat tgaaggagag ggcgaaggga ggccttacga gggcacacag
     3361 actgccaagc tgaaagtgac caagggagga ccactgcctt tcgcttggga tatcctgtct
     3421 cctcagttta tgtacggaag taaggcctat gtcaagcatc ccgctgacat tcctgattac
     3481 aagaaactgt ctttcccaga gggctttaag tgggagagag tgatgaattt tgaagatgga
     3541 ggcctggtga ccgtgacaca ggactcctct ctgcaggatg gcactctgat ctacaaagtc
     3601 aaaatgcgcg gcaccaattt tccacccgat gggcccgtga tgcagaagaa aacaatgggg
     3661 tgggaggcca gcactgaacg gctgtatcct agagacggag tgctgaaggg cgaaatccac
     3721 caggccctga agctgaaaga cggcggccac tacctggtgg agttcaaaac catctacatg
     3781 gccaagaaac cagtgcagct gcccggctat tactatgtgg acaccaagct ggatatcaca
     3841 tcccacaatg aagactacac cattgtggaa cagtatgaga ggtctgaagg acgccaccat
     3901 ctgtttctgt acggcatgga tgagctgtat aagtaaacat ggagatctcg agactagtaa
     3961 ttaagtctca gccaccgtta actgaacatg tcaaaacctg tggagactgt tgagatttga
     4021 tgttctgaaa agataaagcc tataaataaa atgttgccca aatttcctgc ctgatgtttt
     4081 tctttgtctt tgctacatgg ctttgctgct cggatcggct cactctgtgt atgccacgtt
     4141 cactttgtac tctccttctc acggtaggtt tattattttt agatgtgcag ttagtttctg
     4201 tgaaataaca caccacacac tgatattgtc tgtgcattga cttggtgagt gcacattgtt
     4261 tttgatcttg acatatttat atttgattga tcaggtgaac tgtgtgaatc taaagtgctc
     4321 catacagatg ttctgcattg aaaatattct cattttatta gtggaagtga gtgtatgcca
     4381 gcggccgcgg taccctgcag tctagacatt tcagttgacg aagacaaaca aagttctgtt
     4441 gtgactatgg gggggggggg cgcctgggga tggtctcgcc cggggagtaa ttcagggtag
     4501 aaccgccact gcctttagcg acattcactg gccgtcgttt tacaacgtcg tgactgggaa
     4561 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4621 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4681 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     4741 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4801 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4861 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4921 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4981 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5101 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5161 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5221 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5281 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5341 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5401 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5461 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5521 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5581 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5641 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5701 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5761 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5821 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5881 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5941 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6121 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6181 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6241 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6301 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6361 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6421 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6481 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6541 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6601 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6661 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6721 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6781 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6841 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6901 tcagtgagcg aggaagcgga aga
//
