LOCUS       sequence-229560-        7585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For selectively expressing ChR2(H134R) in ASH sensory neurons under 
            the expression of a Psra-6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7585)
  AUTHORS   Hori S, Oda S, Suehiro Y, Iino Y, Mitani S
  TITLE     OFF-responses of interneurons optimize avoidance behaviors depending
            on stimulus strength via electrical synapses.
  JOURNAL   PLoS Genet. 2018 Jun 25;14(6):e1007477. doi: 
            10.1371/journal.pgen.1007477. eCollection 2018 Jun.
  PUBMED    29939997
REFERENCE   2  (bases 1 to 7585)
  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: "PLoS 
            Genet."; date: "2018-06-25"; volume: "14(6):e1007477. doi"; pages: "
            10.1371/journal.pgen.1007477. eCollection 2018 Jun"
COMMENT     Sequence Label: pPD_Psra-6::ChR2(H134R)::mCherry
FEATURES             Location/Qualifiers
     source          1..7585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    92..113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(128..133,134..151,152..158)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    166..182
                     /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     171..193
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     190..206
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     190..206
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2789..3715
                     /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"
     CDS             3728..4438
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3873..3891)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4141..4160
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5360..5382)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5482..5501
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5579..5597)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5665..5769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5770..5838,5839..6630)
                     /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(5988..6007)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6801..7389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7290..7309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7543..7560
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
       61 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      121 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      181 aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat
      241 cttactaact aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca
      301 caacgatgga tacgctaaca acttggaaat gaaataagct tgcatgcctg caggtcgact
      361 ctagaggatc cctgaggtgc atttgcgagg ggcacttcag atttgaaact ttgaattttg
      421 cacctcgcca tgtctataaa aggaggagtt tgataatatc tgacgcaagt agagtacaat
      481 tttattaatc caaataaatt gtgccaattt ttataagatg atataaatga gaaacgggaa
      541 ttgagaatat gatgagtagt ttttaaagtt tgtcattcaa aattgttgtt gcctgccatg
      601 caaatgaaag tcagaagagt tccaaaccag aagaggaaat atgatttttc catgtaacag
      661 tagttgtctt tttttagaac ggcaggccaa acacaaaagt attccattat ttcctgattt
      721 gccttttcta aacaatttcc agtcaagtga taaattcaca tttccagtgc tctgaaaatc
      781 ttgtcaaaaa gtttgtattt cggatgtcag gtgatgtttg taagacatac gttagctggt
      841 ctcttacgta ttccaaaacc gctacaatat gataatcagt caaaatgaaa cactaaagtt
      901 tttgaactga atgatcacag agttgattga gatagataga gttataattt tgaaggaatc
      961 ttacgcatac ggaagcttca aagagtttag gataatttca tagttagagt agacgtgaca
     1021 aagttattaa tatcataatt tgtttttctc gaaattgaat atgactgaaa cgcaagtcag
     1081 agagattcct gaacctaata atctaaaatt ttaattgcta ttttcaatat tgcatttttc
     1141 gattcacgcg taaaggaaga gacaatttca cagcatcgga actatcttcc gatttggaag
     1201 ataatttttt aaagtgtttt ccttattggc tttgaaattt caataaaagt acacctgacg
     1261 aattttcaaa taagtgaaaa atagaagcag acatcgagta aacgtgcata aaataaatgc
     1321 aataagtgct cgttaagtaa tgaaataaag ttagtctaca aattgatcaa ccagtacatg
     1381 actaaaaatt ggtgactgtt ttgttttagt ctttacccaa tttttatttc agcgatttca
     1441 ttttatactc aaatgtgatt tttctacatt ttttcgaatt tagaaaagat cagacgattt
     1501 ccttaaagag gatttctagt ttagtctttg attcaataac ttatagtaca gcttagttat
     1561 aactatagtt cactatgaaa tatcacattt ataatcgttt catttaattg taaggacatt
     1621 tttgcatcaa acgccagttt tctgtcccca tttaattata agttttactc ggccgatgcg
     1681 agatggatgc tcctctcaca caccttaccc cacgccttga caccttattt attgggtttt
     1741 aaattttttt gatattttag agaattataa aggtttccgt taaaaaaaaa catggccttc
     1801 aaggcgagta cgtctgcgtt gtaaactcac aaataaactt aaaaaatatt tttttttgga
     1861 ttgaagcttt atttgaaagg ataactcact cgctatctaa aaattgagaa agttttgtaa
     1921 taacacgaaa tttcatgaag tgcggaaaat ttttgaaaat tggatgaaac tgttcctttc
     1981 acgagtttgg tcgtgtttta ttcttcattc aaataccaat gcaacagtcg gttttaactt
     2041 ttcaaaatta tacttcaagt ttttacaagt gcatatgtaa aagtgttttc tatcttcaca
     2101 ccttcgaaaa cagcaataat aaattaaagc cgcgtgcagc tatcttatag tttggttctc
     2161 tgtattttca aatatcttaa gtgtcacatc aaattcaatg ataatttgga aatcacagaa
     2221 gatctttaat gtgtttatag tctctatgaa ctcatcaatt acaaatatat ttctagtcag
     2281 actattcaca caaaccgcag tttttaatca tccatatctt cttctcatgg aagttctagc
     2341 taacgtcaag tcagactttc acaagtatga gttttatttt gtccaacatt ttatgctctt
     2401 gatgaactca aatatcattt ctctttaaac tatagtttca taagattcag taattttgat
     2461 tctgcagttc agattctaaa tttatccgaa gagaaaacca attgatcaag tctttattca
     2521 atttccgaga aaacgtggtt gagcttaaca cattttagaa tttgtaaacg tgttgttgtt
     2581 tttcaaaacc tcgctaataa tttctcccta gttactcccg taaagtattt tctggaaaat
     2641 ttgcaggttt aatagagttg gtttgaaata ctgcaaatca aaacactttt tatagattaa
     2701 atcaattttg ataatgtttt ctgtcacaca taataaactg cactcgcaga atttaatatt
     2761 tattatttca gattttgcca tgggatccat ggactatggc ggcgctttgt ctgccgtcgg
     2821 acgcgaactt ttgttcgtta ctaatcctgt ggtggtgaac gggtccgtcc tggtccctga
     2881 ggatcaatgt tactgtgccg gatggattga atctcgcggc acgaacggcg ctcagaccgc
     2941 gtcaaatgtc ctgcagtggc ttgcagcagg attcagcatt ttgctgctga tgttctatgc
     3001 ctaccaaacc tggaaatcta catgcggctg ggaggagatc tatgtgtgcg ccattgaaat
     3061 ggttaaggtg attctcgagt tcttttttga gtttaagaat ccctctatgc tctaccttgc
     3121 cacaggacac cgggtgcagt ggctgcgcta tgcagagtgg ctgctcactt gtcctgtcat
     3181 ccttatccgc ctgagcaacc tcaccggcct gagcaacgac tacagcagga gaaccatggg
     3241 actccttgtc tcagacatcg ggactatcgt gtggggggct accagcgcca tggcaaccgg
     3301 ctatgttaaa gtcatcttct tttgtcttgg attgtgctat ggcgcgaaca cattttttca
     3361 cgccgccaaa gcatatatcg agggttatca tactgtgcca aagggtcggt gccgccaggt
     3421 cgtgaccggc atggcatggc tgtttttcgt gagctggggt atgttcccaa ttctcttcat
     3481 tttggggccc gaaggttttg gcgtcctgag cgtctatggc tccaccgtag gtcacacgat
     3541 tattgatctg atgagtaaaa attgttgggg gttgttggga cactacctgc gcgtcctgat
     3601 ccacgagcac atattgattc acggagatat ccgcaaaacc accaaactga acatcggcgg
     3661 aacggagatc gaggtcgaga ctctcgtcga agacgaagcc gaggccggag ccgtgccagg
     3721 cggccgcatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     3781 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     3841 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     3901 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     3961 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     4021 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     4081 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     4141 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     4201 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     4261 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     4321 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     4381 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaag
     4441 atctgaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     4501 ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat
     4561 ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca
     4621 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     4681 tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     4741 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     4801 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     4861 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     4921 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     4981 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     5041 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     5101 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     5161 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     5221 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     5281 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     5341 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5401 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5461 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5521 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     5581 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5641 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5701 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5761 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5821 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5881 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5941 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6001 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6061 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6121 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6181 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6241 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6301 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6361 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6421 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6481 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6541 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6601 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6661 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6721 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6781 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6841 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6901 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6961 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7021 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7081 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7141 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7201 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7261 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7321 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7381 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7441 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7501 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7561 gaagcggaag agcgcccaat acgca
//
