LOCUS       sequence-462091-        8988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pan-neuronal expression of opsin 'ChR2(H134R)', regulated by rab-3 
            promoter and unc-54 3' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8988)
  AUTHORS   Sharma AK, Randi F, Kumar S, Dvali S, Leifer AM
  TITLE     TWISP: a transgenic worm for interrogating signal propagation in 
            Caenorhabditis elegans.
  JOURNAL   Genetics. 2024 Jul 8;227(3):iyae077. doi: 10.1093/genetics/iyae077.
  PUBMED    38733622
REFERENCE   2  (bases 1 to 8988)
  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: "Genetics.";
            date: "2024-07-8"; volume: "227(3):iyae077. doi"; pages: " 
            10.1093/genetics/iyae077"
COMMENT     Sequence Label: pAS3-rab-3P::AI::ChR2(H134R)
FEATURES             Location/Qualifiers
     source          1..8988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4606..5532
                     /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(6450..6472)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6572..6591
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6669..6687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6755..6859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6860..6928,6929..7720)
                     /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(7078..7097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7891..8479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8380..8399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8633..8650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8767..8788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8803..8808,8809..8826,8827..8833)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8841..8857
                     /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     8846..8868
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8865..8881
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8865..8881
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gctaacaact tggaaatgaa ataagcttgc atgcctgcag gcgagttttg actggctttc
       61 aaaaactgag tgattactaa ttatttttga aaaaattaac agttgtgata tgatatttag
      121 acatttttgc aatatataat ttgtttgaaa aaaaaacatt ttttaatggt cttcgaagct
      181 ataatagttt ttcgtaaaat tttaaataaa acagctgaca gatatccaca ttcaataatt
      241 gaaagtatgc agttctcttt ttttttgtca aatagaaaaa aagtaacaga tggtcagata
      301 taatctaggt taatctgact ctggtcatcg atgaactcat tttcatcata atcattcttc
      361 tttcaaatgg tctgttcgga attcgaatat tcaataaatg gatatcttat tagatgctcc
      421 ggatttattc caccaaactt ccaagacgta agttcaattt aaacaaaatt actatagctt
      481 gaaaaaatga gaaaaaacga gcagataaaa aaataacaga aaaaatccat aaaaatgacg
      541 agggttccgt gcgaaaatcg tcattgtttt tggcttgcat cacacttttc cgcgagtcag
      601 ccacactgtt ttatcgatta ttatttttct gatttcagaa accggaaact tacaactgag
      661 atgctttcat tttgattttt ttaaatttag ttttgatttc agcgatttgt ttgaatataa
      721 ttatgcatga gttggctttt gcatttttag ataaataagt tcacatgctc ggacaaattg
      781 tgtggtttaa tctgtgaaag ttttgaatct tctgtttgaa aaaaaaaaca tttttaattt
      841 gtatatacga gtgaattggt taggccctgg ttacgttttc cagcgaattc attcaaattt
      901 cttatttttt ctattttttc aattaaataa tttagttctc tatcacatgt gtatgatcaa
      961 aatttcattc ctaatcgtaa gcaaatttag gctcatccgt agtataattt tgttatgaag
     1021 ggcttcacag taaaagctca aactccaaca tttagcaact agcaaccaac aaaaaaccaa
     1081 agaaatgtca gttttgaaaa gtcgttattt ctttgtcttc tcacaattca cacatcatct
     1141 tctcgttttc ctccgttttt ttgtcgtgct tccttctttt ctttacacat ttttctaacc
     1201 tttttttaag tctatcctct cctaatcctt tgtactctct aatcagtttt gctggttgtg
     1261 ctgcacaatt tttatcagat ttatgcgcga aacaaaagcc tctcgcggag agctccgcca
     1321 aatccggtct tcttttttcc ttgcctatcg gatttcacaa acattcagtt tgtcacttga
     1381 cgatgtgttg gaggcattac tactaggaag tgttattcat attttttatt cgtttctttt
     1441 tggtgactaa acgaaatatt ccgaacgttt tcctgatttc aggcggcgga aacgaagaag
     1501 accacgaata atcaacaggc tgccattgca tccgccagaa gtcgagtgag ttctttgaaa
     1561 atctttatta gagttatttg tgacaatttg ccactttttt tttaatgttt aatgaatctg
     1621 taacgaaatt aataccaagg gtgtccaaca gaactatctg agcggttatt taatttttag
     1681 atagtatatt tcttccgatc gtaacataca tattttcaag catagtactg tgatttaagt
     1741 tacagttttc caatcagtct tgattttaat aaaaactttt caactgcaaa aatgcctctg
     1801 aagtattttt ctatattttt atagttgaat aattttttga ttggcacaaa gaccgccgag
     1861 atatgagccg ttaaagtaaa gctgcatgga ttcaatatgg tttgtatttg attggagtca
     1921 cttacatgga aatatttgta tttaaatctc atattttaaa acaagacaaa caccaaatta
     1981 aatttttatt aatctgaaca tttttcaatc gatttggtaa acaaattatt agatgtttcc
     2041 ctcaaaaatt gtaacaattc ctaaattgtg gcacttataa taacatcgtc ctgaattctc
     2101 aaaaaaaacc gaatgctaag agagatttcg tttaactggg acctgtaact gaattttcaa
     2161 atattggcag tttttgataa actgggaggg aaatcaaaaa ccaaaactac agtacccctc
     2221 gtgttataat aaattgtcat tcactccttt tttgttccca acatttgctc ccatctcctg
     2281 acgagaaaga acaattttat gagcacggga acccgtgggc gtccatccgt tcctcctgtg
     2341 caatttcagc ccccttcctc tacattctct gtgttcccgt ttttatcgtt ttatgggtat
     2401 cgggaacgga cagatcgctt ccatttgctt ctattccgtc ctcacaagac gtcccactaa
     2461 ctgattttct tctgatttct tttcactgaa aatgggaaag aaagaaaaag acgatgaatc
     2521 gacgacttct tctcaggaga aggtgaagaa gaaggataag gagaaagttc atctaaggct
     2581 gaaagataaa gtaaaaaaac ttagaaaatc taagtcgatt gcagtatctt ctccagctga
     2641 ttatatgact tgtcagtgtg aaccatgcaa aaaagtagct ctcgaggtaa taatttttaa
     2701 actcaatacg tagttaagtt tgaactcctt ttaatttact tatcatctaa tttaccgtca
     2761 caatagtact tccattagca attattttaa aattacatgc atagcttcaa tgttcgtttt
     2821 tttgcgttct catatcgtat gttgtctgcc tctcgaatac tgactgtccg actagaatgt
     2881 cccggtagga ttactgtacg gtcactggga ataaactgtt tctgaaatca atctgaaatt
     2941 ccgtagatct tcttggttca atttcttatt cctcaatctt tggacaacgt caccttcaga
     3001 gtcgtcgttg agtatcccat cattaactag tcaatctttc tctctttttc atttccccca
     3061 acttttctgc atattttttg acgacgacga cctcgacggc aacgagaata catttatcta
     3121 ttttttagta tagcatatat acattttcta cttttctcta acccatttca gacgtataga
     3181 ttttggtttt cgaatgcaac tttgagattt ttcaaagatc ttcagatggg agcagtggac
     3241 tgtctatttg atattcttaa aatgtactct ataaaagatg aaactgtaat gaccaattcc
     3301 ctacataaat atttttaact aattggtcgg atatttgggg atcagagaag tacttgtttt
     3361 tgcgatatta ataatgatat atttttctag taaaaaaatg ttaagcgatt tgtgaataat
     3421 taagttggct tgaaaaagat tttttgatag ctatgtattt tgttaattta aatcagattt
     3481 taaaacttta ttattgacta tttatatata ttatatatta tatattttca tggtttaatc
     3541 aaaatctatt aaattatttc taagatctct aaatatcgaa ttctctaaaa agcttcatca
     3601 acaaaagcat ttcaattatc taacacaatg agaataattc aagaactgaa atagaaagaa
     3661 atctagaaaa aaaagagagt gaggtgccct attaaatcgg tgctctcctc tctaacagaa
     3721 gcgcctacac gcacccaaga gagctctttt tctttctttt ttcggagcac acatacacgc
     3781 atacactcct cagaagtgta tagagaaaat gggagaagaa gcgaaagaag acgaagaagc
     3841 ggttgtgatg tatgcatcgg gcgtttctat ccctctccgt gagcaacgag ctagtcaacc
     3901 catcgatcca tttttgtgag cacacacaga gagagactca aacacactca cagcttcaca
     3961 ctgagctcca agagctctgt agagagaagc cagagtatgc tttcctcgat tttcctaacg
     4021 tcttcttctt ctgctttttt ctttttccca acgagttgta gtccactaac ttcaattttt
     4081 cctccagaga aagagtttta gagagacagc tgataaatac actagagaaa taaaaagaat
     4141 gataaaaatt tgctatttta atcagttccc tctcgtttct ctaagttttg ttgttgggtt
     4201 gggaaattaa aattggaaaa tggaaataac gtttttttct gtctaatctt gaattctcca
     4261 gacatttctt atttcctttt ttgttgcaag tttttcttcc ttctctaatt tttaatcaaa
     4321 gatatctcgc tatgaacttt gaatttttga tattttgatt ctataaatac ataaagtaat
     4381 ctaaatgctc ttttaaaata aatctacagt agccctattt tcagggatcc ccgggattgg
     4441 ccaaaggacc caaaggtatg tttcgaatga tactaacata acatagaaca ttttcaggag
     4501 gacccttgga gggtaccgag ctcagaaaaa atgttttctg tcacacataa taaactgcac
     4561 tcgcagaatt taatatttat tatttcagat tttgccatgg gatccatgga ctatggcggc
     4621 gctttgtctg ccgtcggacg cgaacttttg ttcgttacta atcctgtggt ggtgaacggg
     4681 tccgtcctgg tccctgagga tcaatgttac tgtgccggat ggattgaatc tcgcggcacg
     4741 aacggcgctc agaccgcgtc aaatgtcctg cagtggcttg cagcaggatt cagcattttg
     4801 ctgctgatgt tctatgccta ccaaacctgg aaatctacat gcggctggga ggagatctat
     4861 gtgtgcgcca ttgaaatggt taaggtgatt ctcgagttct tttttgagtt taagaatccc
     4921 tctatgctct accttgccac aggacaccgg gtgcagtggc tgcgctatgc agagtggctg
     4981 ctcacttgtc ctgtcatcct tatccgcctg agcaacctca ccggcctgag caacgactac
     5041 agcaggagaa ccatgggact ccttgtctca gacatcggga ctatcgtgtg gggggctacc
     5101 agcgccatgg caaccggcta tgttaaagtc atcttctttt gtcttggatt gtgctatggc
     5161 gcgaacacat tttttcacgc cgccaaagca tatatcgagg gttatcatac tgtgccaaag
     5221 ggtcggtgcc gccaggtcgt gaccggcatg gcatggctgt ttttcgtgag ctggggtatg
     5281 ttcccaattc tcttcatttt ggggcccgaa ggttttggcg tcctgagcgt ctatggctcc
     5341 accgtaggtc acacgattat tgatctgatg agtaaaaatt gttgggggtt gttgggacac
     5401 tacctgcgcg tcctgatcca cgagcacata ttgattcacg gagatatccg caaaaccacc
     5461 aaactgaaca tcggcggaac ggagatcgag gtcgagactc tcgtcgaaga cgaagccgag
     5521 gccggagccg tgccaggcgg ccgctaagcg ccggtcgcta ccattaccaa cttgtctggt
     5581 gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa
     5641 cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta
     5701 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     5761 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg
     5821 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     5881 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     5941 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     6001 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     6061 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     6121 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     6181 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     6241 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     6301 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     6361 acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt
     6421 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6481 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6541 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6601 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     6661 ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6721 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6781 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6841 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6901 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6961 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7021 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7081 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7141 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7201 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7261 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7321 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7381 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7441 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7501 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7561 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7621 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7681 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7741 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7801 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7861 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7921 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7981 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     8041 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8101 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8161 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8221 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     8281 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8341 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8401 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8461 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8521 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     8581 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     8641 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     8701 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     8761 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     8821 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     8881 ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta
     8941 aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatac
//
