LOCUS       sequence-229561-        6164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For selectively expressing ChR2(H134R) in AiB interneurons under the
            expression of a inx-1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  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 6164)
  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_Pinx-1::ChR2(H134R)::mCherry
FEATURES             Location/Qualifiers
     source          1..6164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     360..377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    494..515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(530..535,536..553,554..560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    568..584
                     /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     573..595
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     592..608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     592..608
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1770..2696
                     /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             2709..3419
                     /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(2854..2872)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3122..3141
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4341..4363)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4463..4482
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4560..4578)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4646..4750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4751..4819,4820..5611)
                     /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(4969..4988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5782..6164,1..206)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
       61 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      301 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
      361 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      481 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      541 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      601 gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa ctaactattc
      661 tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg gatacgctaa
      721 caacttggaa atgaaataag cttgttacta aatagattca aaaattattt tctgtgcttt
      781 tcacaaatac actttatttt aggtaatcat attttaatta aacacgcggg aaatttttta
      841 gggtaaactt tgaagttttt tttgtcgcgg atatcagatt cataactgtc tatgtaccat
      901 aaattacgac ttttgaatat ttgctgttca aagtcaatgt ataaattaaa tttaagtttc
      961 tttcgtcata catctcaaaa atagctaaaa tgcgtttaac aaatgttata cctaattttc
     1021 aatcataact tggagagtat gaatgaattc aaacgtttcc ttgaggtgat atccttttgg
     1081 tacttgtata ttatcgaggc acagtttgaa aataaattaa attttaattc atttgatgat
     1141 tttgttttct cttgaggctt aaaaatgata aacggtacaa aactacaaaa aaactccata
     1201 agtctttatt ttcttaattt ttgaaatttt atttcaaaat gcacaagatc catttatcat
     1261 atttatagtt cttcatcttc ttttttttaa tatcctcttt ttgtgtttta ttcggtccat
     1321 acgtgctctt caactttttt ttcatttttg actttaaata agaaaagatg gcaataaaaa
     1381 atttggccga agagggcgat ggacggatga aaatctacta aaaggattat aactcaattt
     1441 gatatgctct tcgggaggta tcctcctgac gagatggaaa agaagaagaa gaagaagaag
     1501 aagcttgtca tcgtttcatc ggcaaagaga cgggtggaca ttagaccacg ccccacaggg
     1561 aaacctttcg agtttcatcc acctctgtct gtttaacata tttgtttttt tctagctcta
     1621 tttttcttcc gcttctactg cgtacttttg cataattcta tttctaagtc tgatcattat
     1681 aagtatcatc ctgaacatcg cacactaaac atcctcggcc ggcacggatg aagtagtttt
     1741 cattgcagtt cttgtccgcc ggaggatcca tggactatgg cggcgctttg tctgccgtcg
     1801 gacgcgaact tttgttcgtt actaatcctg tggtggtgaa cgggtccgtc ctggtccctg
     1861 aggatcaatg ttactgtgcc ggatggattg aatctcgcgg cacgaacggc gctcagaccg
     1921 cgtcaaatgt cctgcagtgg cttgcagcag gattcagcat tttgctgctg atgttctatg
     1981 cctaccaaac ctggaaatct acatgcggct gggaggagat ctatgtgtgc gccattgaaa
     2041 tggttaaggt gattctcgag ttcttttttg agtttaagaa tccctctatg ctctaccttg
     2101 ccacaggaca ccgggtgcag tggctgcgct atgcagagtg gctgctcact tgtcctgtca
     2161 tccttatccg cctgagcaac ctcaccggcc tgagcaacga ctacagcagg agaaccatgg
     2221 gactccttgt ctcagacatc gggactatcg tgtggggggc taccagcgcc atggcaaccg
     2281 gctatgttaa agtcatcttc ttttgtcttg gattgtgcta tggcgcgaac acattttttc
     2341 acgccgccaa agcatatatc gagggttatc atactgtgcc aaagggtcgg tgccgccagg
     2401 tcgtgaccgg catggcatgg ctgtttttcg tgagctgggg tatgttccca attctcttca
     2461 ttttggggcc cgaaggtttt ggcgtcctga gcgtctatgg ctccaccgta ggtcacacga
     2521 ttattgatct gatgagtaaa aattgttggg ggttgttggg acactacctg cgcgtcctga
     2581 tccacgagca catattgatt cacggagata tccgcaaaac caccaaactg aacatcggcg
     2641 gaacggagat cgaggtcgag actctcgtcg aagacgaagc cgaggccgga gccgtgccag
     2701 gcggccgcat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     2761 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2821 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2881 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     2941 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     3001 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     3061 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     3121 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     3181 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     3241 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     3301 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     3361 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaaa
     3421 gatctgaatt ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat
     3481 aataggggcc gctgtcatca gagtaagttt aaactgagtt ctactaacta acgagtaata
     3541 tttaaatttt cagcatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac
     3601 atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaac
     3661 ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt
     3721 agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc
     3781 cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt
     3841 ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc
     3901 atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca
     3961 tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca
     4021 agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc cctattgttt
     4081 gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat gagcacgatg
     4141 caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt agaagttgat
     4201 aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa tacattccca
     4261 atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg tctcgcgcgt
     4321 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4381 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4441 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4501 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg
     4561 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4981 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5281 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5341 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6121 gcccagcttg gagcgaacga cctacaccga actgagatac ctac
//
