LOCUS       sequence-236922-        6860 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Zebrafish Tol1 construct for UAS-mediated expression of the 
            optogenetic opsin CoChR fused to the red fluorescent protein 
            tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6860)
  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 6860)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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:CoChR-tdTomato
FEATURES             Location/Qualifiers
     source          1..6860
                     /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"
     CDS             1564..2427
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     primer_bind     complement(4464..4481)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4464..4480)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4473..4495)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4689..4708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4808..4830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4868..4886)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4954..5058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5059..5127,5128..5919)
                     /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(5277..5296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6090..6678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6579..6598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6832..6849
                     /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 accatgctgg gaaacggcag cgccattgtg cctatcgacc agtgcttttg cctggcttgg
     1621 accgacagcc tgggaagcga tacagagcag ctggtggcca acatcctcca gtggttcgcc
     1681 ttcggcttca gcatcctgat cctgatgttc tacgcctacc agacttggag agccacttgc
     1741 ggttgggagg aggtctacgt ctgttgcgtc gagctgacca aggtcatcat cgagttcttc
     1801 cacgagttcg acgaccccag catgctgtac ctggctaacg gacaccgagt ccagtggctg
     1861 agatacgcag agtggctgct gacttgtccc gtcatcctga tccacctgag caacctgacc
     1921 ggcctgaagg acgactacag caagcggacc atgaggctgc tggtgtcaga cgtgggaacc
     1981 atcgtgtggg gagctacaag cgccatgagc acaggctacg tcaaggtcat cttcttcgtg
     2041 ctgggttgca tctacggcgc caacaccttc ttccacgccg ccaaggtgta tatcgagagc
     2101 taccacgtgg tgccaaaggg cagacctaga accgtcgtgc ggatcatggc ttggctgttc
     2161 ttcctgtctt ggggcatgtt ccccgtgctg ttcgtcgtgg gaccagaagg attcgacgcc
     2221 atcagcgtgt acggctctac cattggccac accatcatcg acctcatgag caagaattgt
     2281 tggggcctgc tgggacacta tctgagagtg ctgatccacc agcacatcat catctacggc
     2341 gacatccgga agaagaccaa gatcaacgtg gccggcgagg agatggaagt ggagaccatg
     2401 gtggaccagg aggacgagga gacagtggcg gcaccggtag tagcagtgag taagggcgag
     2461 gaagtgatca aagagttcat gcggtttaag gtgagaatgg aaggaagcat gaacggccac
     2521 gagttcgaaa ttgagggaga aggagaggga cggccctacg agggcaccca gacagccaag
     2581 ctgaaagtga caaagggcgg gcctctgcca ttcgcttggg acatcctgag cccacagttt
     2641 atgtacggct ccaaggccta tgtgaaacat ccagctgaca ttcccgatta taagaaactg
     2701 agcttccccg aggggtttaa gtgggaaaga gtgatgaact tcgaggacgg aggcctggtg
     2761 actgtgaccc aggacagctc cctgcaggat gggaccctga tctacaaggt gaaaatgaga
     2821 gggacaaatt ttccccctga tggacctgtg atgcagaaga aaactatggg atgggaggcc
     2881 tccaccgaaa ggctgtatcc acgcgacggg gtgctgaaag gagaaatcca ccaggctctg
     2941 aagctgaaag atgggggaca ttacctggtg gagttcaaga caatctacat ggccaagaaa
     3001 cctgtgcagc tgccaggcta ctattacgtg gacacaaaac tggatatcac ttcacacaac
     3061 gaggactaca ctattgtgga gcagtatgaa cggagcgagg ggagacacca tctgttcctg
     3121 ggccatggga ctggaagtac cggctcaggg tctagtggaa ccgcctcaag cgaggataac
     3181 aatatggctg tgatcaaaga gttcatgagg tttaaggtgc gcatggaggg cagcatgaat
     3241 gggcacgaat ttgagattga aggagagggc gaagggaggc cttacgaggg cacacagact
     3301 gccaagctga aagtgaccaa gggaggacca ctgcctttcg cttgggatat cctgtctcct
     3361 cagtttatgt acggaagtaa ggcctatgtc aagcatcccg ctgacattcc tgattacaag
     3421 aaactgtctt tcccagaggg ctttaagtgg gagagagtga tgaattttga agatggaggc
     3481 ctggtgaccg tgacacagga ctcctctctg caggatggca ctctgatcta caaagtcaaa
     3541 atgcgcggca ccaattttcc acccgatggg cccgtgatgc agaagaaaac aatggggtgg
     3601 gaggccagca ctgaacggct gtatcctaga gacggagtgc tgaagggcga aatccaccag
     3661 gccctgaagc tgaaagacgg cggccactac ctggtggagt tcaaaaccat ctacatggcc
     3721 aagaaaccag tgcagctgcc cggctattac tatgtggaca ccaagctgga tatcacatcc
     3781 cacaatgaag actacaccat tgtggaacag tatgagaggt ctgaaggacg ccaccatctg
     3841 tttctgtacg gcatggatga gctgtataag taaacatgga gatctcgaga ctagtaatta
     3901 agtctcagcc accgttaact gaacatgtca aaacctgtgg agactgttga gatttgatgt
     3961 tctgaaaaga taaagcctat aaataaaatg ttgcccaaat ttcctgcctg atgtttttct
     4021 ttgtctttgc tacatggctt tgctgctcgg atcggctcac tctgtgtatg ccacgttcac
     4081 tttgtactct ccttctcacg gtaggtttat tatttttaga tgtgcagtta gtttctgtga
     4141 aataacacac cacacactga tattgtctgt gcattgactt ggtgagtgca cattgttttt
     4201 gatcttgaca tatttatatt tgattgatca ggtgaactgt gtgaatctaa agtgctccat
     4261 acagatgttc tgcattgaaa atattctcat tttattagtg gaagtgagtg tatgccagcg
     4321 gccgcggtac cctgcagtct agacatttca gttgacgaag acaaacaaag ttctgttgtg
     4381 actatggggg gggggggcgc ctggggatgg tctcgcccgg ggagtaattc agggtagaac
     4441 cgccactgcc tttagcgaca ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4501 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4561 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4621 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4681 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4741 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4801 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4861 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4921 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4981 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5041 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5101 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5161 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5221 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5281 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5341 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5401 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5461 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5521 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5581 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5641 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5701 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5761 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5821 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5881 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5941 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6001 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6061 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6721 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6781 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6841 gtgagcgagg aagcggaaga
//
