LOCUS Exported 6164 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION For selectively expressing ChR2(H134R) in AiB interneurons under the expression of a inx-1 promoter. ACCESSION . VERSION . KEYWORDS pPD_Pinx-1::ChR2(H134R)::mCherry 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6164 /organism="synthetic DNA construct" /mol_type="other DNA" 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 530..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 4751..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 5782..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 //