LOCUS Exported 5629 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION slow cycling (open for minutes) step-function artificial anion conducting channelrhodopsin (aACR). High light sensitivity. Activation with green light, inactivation max with 625 nm (accelarates closure to ms). Not recommended for in vivo use. Codon optimized for mammalian expression.. ACCESSION . VERSION . KEYWORDS p-mCherry-C1-Aurora_CA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5629) AUTHORS Wietek J, Rodriguez-Rozada S, Tutas J, Tenedini F, Grimm C, Oertner TG, Soba P, Hegemann P, Wiegert JS TITLE Anion-conducting channelrhodopsins with tuned spectra and modified kinetics engineered for optogenetic manipulation of behavior. JOURNAL Sci Rep. 2017 Nov 2;7(1):14957. doi: 10.1038/s41598-017-14330-y. PUBMED 29097684 REFERENCE 2 (bases 1 to 5629) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5629 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 119..422 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 423..626 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 577..597 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1594..2301 /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(1736..1754) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2004..2023 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2475..2596 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2512..2531) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2566..2585 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2603..3058) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2740..2761) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2952..2971 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3085..3189 /gene="bla" /label=AmpR promoter promoter 3191..3548 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3214..3234) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3399..3534 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3461..3480 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3583..4377 /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(3637..3656) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4247..4266 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4565..4584) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4609..4656 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4985..5573 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5474..5493 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta 61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgccaca 661 atggattacg gcggagccct gtcagcagtc ggcctgtttc agacctcata taccctggag 721 aataacggca gcgtcatctg tatcccaaat aacgggcagt gtttctgcct ggcatggctg 781 aagagcaacg gcaccaatgc agagaagctg gcagccaata tcctgcagtg ggtgtcattt 841 gccctgtctg tggcatgtct gggatggtac gcctatcagg catggagagc cacttgtgga 901 tgggagaacg tgtatgtcgc tctgattcag atgatgaaga gcatcattga ggccttccat 961 tcctttgatt caccagctac tctgtggctg tcaagtggca acggggtcag gtggatgaga 1021 tatgggagct ggctgctgac cgcccccgtg attctgatcc acctgagcaa cctgactggc 1081 ctgaaggacg attacagcaa gcgcactatg gggctgctgg tctcagacgt gggatgtatc 1141 gtgtggggcg ccacaagcgc aatgtgcaca ggctggacca agatcctgtt tttcctgatc 1201 agcctgtctt acggcatgta cacttacttc catgccgcaa aggtgtacat cgaggctttc 1261 cacaccgtgc caaagggact gtgcagacag ctggtgcggg caatggcttg gctgttcttt 1321 gtgagctggg ggatgtttcc agtcctgttt ctgctgggac ccgagggctt cgggcacatc 1381 agcagatacg ggtccaacat tggccacagc attctggatc tgattgccaa gcagatgtgg 1441 ggagtgctgg gaaactacct gagggtgaaa attcacagcc atatcctgct gtacggagat 1501 atccgcaaga agcagaaaat caccattgca gggcaggaaa tggaagtgga aactctggtc 1561 gccgaggagg aagacgctgg ggctgtaccg gtggtgagca agggcgagga ggataacatg 1621 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac 1681 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag 1741 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc 1801 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg 1861 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg 1921 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc 1981 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc 2041 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg 2101 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag 2161 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac 2221 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc 2281 atggacgagc tgtacaagta agagctcaag cttcgaattc tgcagtcgac ggtaccgcgg 2341 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg 2401 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg 2461 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 2521 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 2581 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta 2641 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 2701 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 2761 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 2821 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 2881 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 2941 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 3001 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca 3061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 3121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 3181 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa 3241 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3301 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3361 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 3421 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3481 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 3541 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3601 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3661 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3721 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg 3781 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3841 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3901 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3961 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4021 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4081 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg 4141 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4201 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4261 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4321 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4381 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4441 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4501 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc 4561 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 4621 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg 4681 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt 4741 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg 4801 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac 4861 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 4921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 4981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 5041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 5101 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 5161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 5221 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 5281 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 5341 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 5401 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 5461 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 5521 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 5581 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctca //