LOCUS Exported 5915 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Anion-conducting Channelrhodopsin with near-complete desensitization in continuous light. Codon-optimized for mammalian expression (human/mouse).. ACCESSION . VERSION . KEYWORDS p-mCherry-C1-MerMAID4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5915) AUTHORS Oppermann J, Fischer P, Silapetere A, Liepe B, Rodriguez-Rozada S, Flores-Uribe J, Peter E, Keidel A, Vierock J, Kaufmann J, Broser M, Luck M, Bartl F, Hildebrandt P, Simon Wiegert J, Beja O, Hegemann P, Wietek J TITLE MerMAIDs: a family of metagenomically discovered marine anion-conducting and intensely desensitizing channelrhodopsins. JOURNAL Nat Commun. 2019 Jul 25;10(1):3315. doi: 10.1038/s41467-019-11322-6. PUBMED 31346176 REFERENCE 2 (bases 1 to 5915) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5915 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 45..348 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 349..552 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 503..523 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1806..2513 /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(1948..1966) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2216..2235 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2687..2808 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2724..2743) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2778..2797 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2815..3270) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2952..2973) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3164..3183 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3297..3401 /gene="bla" /label=AmpR promoter promoter 3403..3760 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3426..3446) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3611..3746 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3673..3692 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3795..4589 /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(3849..3868) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4459..4478 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4777..4796) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4821..4868 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5197..5785 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5686..5705 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 121 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 181 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 241 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 301 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 541 ataagcagag ctggtttagt gaaccgtcag atccgctagc caccatgact atcgtgctgt 601 cttccctgat tgtgattcgc ttcgtgtttt tcgctggcaa taatgtgctg ctgagatgta 661 tgaggctgtg gtactatagg aatatcctgc tgcctgccaa gaacaccaat ctgatcacaa 721 tgtctgacga gagcacaatg gtgccaatct acgccacctt ccagctgttt gccgtgctga 781 tcaccgcagc caccctggtg ttctacgtga gccagctgag aaacggcaat aagggcgatt 841 gggagcccat ctacgtgtct ggcgtggagt ttatcgccta tctgatggcc ctggtgctgc 901 ctgacggcta cgccttccgg agctttcagg atggcgccgg catcatcaca gtgccagtgt 961 tccgctacgt gagctggctg gccacatgcc ccatcatcct gaaggtgctg gtgaccgtga 1021 tctctcacga ggacaacatc gaccaggatg tgatcctgag gctgatgctg ctgatcacat 1081 ggatcgagct gctgggcttt atgggctcta tgtacgtggg catgatcaag gtgttcttta 1141 tcgccctggc cgtgatcctg tgcatctata tgtactatat catctgtatg acctggaaca 1201 agaatcacca cgagtctctg agcctgtcct acaggaagga gagattcgag ctgatgagcc 1261 tgctgatggt gtcctggatt ctgtttccca tcctgtatat cgtgggccct gagtgtttcg 1321 gcctgatctc caatcagttt tctatcatcg gccacgtgat cggcgacatc atcagcaaga 1381 acctgtgggg cctgatggcc tggaagctgc ggctgaagat caaggatggc cagcctatgg 1441 agcacgtgaa tccactgaag atcgtgtcca acgacggcac caatacaatg gagaactacc 1501 cactggataa gatcatccag acctggaagg gccgggagta cgatcgcgag tctatctatg 1561 agaacggccc tagctcccca aagaccccca gcgagacacc ccctaagacc ccacccaata 1621 agatgcactt ccacaacttt ctgctggagc actacatgaa gaccagggag acagcctata 1681 atgagagaaa ggacgcaggc gtgtccggag gcgatctgcc cttccagcag gcccgcagcc 1741 ctaaaggacc catccagtat gaagtcatca agcagtcaag cgtgaacgag tatgccggac 1801 cggtggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca 1861 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg 1921 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc 1981 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc 2041 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc 2101 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg 2161 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg 2221 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg 2281 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg 2341 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg 2401 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg 2461 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taagagctca 2521 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact 2581 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca 2641 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc 2701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 2761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg 2821 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 2881 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 2941 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 3001 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 3061 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 3121 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 3181 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 3241 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg 3301 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 3361 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca 3421 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 3481 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 3541 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3601 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3661 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 3721 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat 3781 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3841 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3901 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3961 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4021 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4081 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4141 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4201 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4261 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4321 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4381 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4441 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4501 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4561 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 4621 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4681 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4741 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc 4801 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc 4861 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag 4921 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc 4981 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg 5041 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 5101 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 5161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 5221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 5281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 5341 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 5401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 5461 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 5521 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 5581 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 5641 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 5701 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 5761 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 5821 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 5881 ccgtattacc gccatgcatt agttattaat agtaa //