LOCUS Exported 5660 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Historical plasmid. For inhibition of neuronal activity, #66709 is a light-gated chloride channel with improved anion selectivity (iChloC). ACCESSION . VERSION . KEYWORDS p-mCherry-N1-slowChloC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5660) AUTHORS Wietek J, Wiegert JS, Adeishvili N, Schneider F, Watanabe H, Tsunoda SP, Vogt A, Elstner M, Oertner TG, Hegemann P TITLE Conversion of Channelrhodopsin into a Light-Gated Chloride Channel. JOURNAL Science. 2014 Mar 27. PUBMED 24674867 REFERENCE 2 (bases 1 to 5660) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5660 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 609..665 /label=MCS /note="multiple cloning site" regulatory 673..682 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 679..1605 /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 NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIRMVKVILEFFFEFKN PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSNIGCIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGAV" CDS 1618..2325 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(1760..1778) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2028..2047 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2448..2569 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2485..2504) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2539..2558 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2576..3031) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2713..2734) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2925..2944 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3058..3162 /gene="bla" /label=AmpR promoter promoter 3164..3521 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3187..3207) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3372..3507 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3434..3453 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3556..4350 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3610..3629) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4220..4239 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4538..4557) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4582..4629 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4958..5546 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5447..5466 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg 661 gatccacccg gtgccaccat ggactatggc ggcgctttgt ctgccgtcgg acgcgaactt 721 ttgttcgtta ctaatcctgt ggtggtgaac gggtccgtcc tggtccctga ggatcaatgt 781 tactgtgccg gatggattga atctcgcggc acgaacggcg ctcagaccgc gtcaaatgtc 841 ctgcagtggc ttgcagcagg attcagcatt ttgctgctga tgttctatgc ctaccaaacc 901 tggaaatcta catgcggctg ggaggagatc tatgtgtgcg ccattagaat ggttaaggtg 961 attctcgagt tcttttttga gtttaagaat ccctctatgc tctaccttgc cacaggacac 1021 cgggtgcagt ggctgcgcta tgcagagtgg ctgctcactt gtcctgtcat ccttatccac 1081 ctgagcaacc tcaccggcct gagcaacgac tacagcagga gaaccatggg actccttgtc 1141 tcaaacatcg ggtgtatcgt gtggggggct accagcgcca tggcaaccgg ctatgttaaa 1201 gtcatcttct tttgtcttgg attgtgctat ggcgcgaaca cattttttca cgccgccaaa 1261 gcatatatcg agggttatca tactgtgcca aagggtcggt gccgccaggt cgtgaccggc 1321 atggcatggc tgtttttcgt gagctggggt atgttcccaa ttctcttcat tttggggccc 1381 gaaggttttg gcgtcctgag cgtctatggc tccaccgtag gtcacacgat tattgatctg 1441 atgagtaaaa attgttgggg gttgttggga cactacctgc gcgtcctgat ccacgagcac 1501 atattgattc acggagatat ccgcaaaacc accaaactga acatcggcgg aacggagatc 1561 gaggtcgaga ctctcgtcga agacgaagcc gaggccggag ccgtgccagc ggccgccgtg 1621 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac 1681 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc 1741 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc 1801 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc 1861 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg 1921 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag 1981 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag 2041 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg 2101 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc 2161 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc 2221 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc 2281 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaagcgg ccgcgactct 2341 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac 2401 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg 2461 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2521 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt 2581 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 2641 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 2701 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 2761 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca 2821 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga 2881 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa 2941 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc 3001 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc 3061 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 3121 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc 3181 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 3241 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 3301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 3361 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 3421 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 3481 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga 3541 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 3601 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 3661 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3721 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 3781 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 3841 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 3901 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 3961 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 4021 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 4081 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 4141 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 4201 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 4261 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 4321 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 4381 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4441 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 4501 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac 4561 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt 4621 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga 4681 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt 4741 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag 4801 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 4861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 4921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 4981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 5041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 5101 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 5161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 5221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 5281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 5341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 5401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 5461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 5521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 5581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 5641 accgtattac cgccatgcat //