LOCUS Exported 5489 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-MerMAID1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5489) 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 5489) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5489 /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" regulatory 579..588 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1380..2087 /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(1522..1540) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1790..1809 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2261..2382 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2298..2317) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2352..2371 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2389..2844) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2526..2547) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2738..2757 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2871..2975 /gene="bla" /label=AmpR promoter promoter 2977..3334 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3000..3020) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3185..3320 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3247..3266 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3369..4163 /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(3423..3442) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4033..4052 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4351..4370) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4395..4442 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4771..5359 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5260..5279 /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 caccatggtc cctatctacg 601 caaccttcca gctgttcgcc gtcctgatta ccgccgccac tctggtcttt tatgtgagcc 661 agctgagaaa tggcaacaag ggcgactggg agcccatcta cgtgtccggc gtggagttca 721 tcgcctatct gatggccctg gtgctgcctg atggctacgc cttcaggtcc tttcaggacg 781 attctggcat catcaccgtg cccgtgttca ggtatgtgag ctggctggcc acctgtccaa 841 tcatcctgaa ggtgctggtg acagtgatct cccacgagga caatatcgac caggatgtga 901 tcctgaggct gatgctgctg atcacatgga tcgagctgct gggcttcatg ggctctatgt 961 acgtgggcat gctgaaggtg ttttgcatcg ccctggccat catcctgtgc atctatatgt 1021 actatatcat ctgcaccaca tggaacaaga atcaccacga gagcctgctg ctgtcctacc 1081 ccaaggagcg gttcgagctg atggccctgc tgatggtgtc ttggatcgtg tttccaatcc 1141 tgtatatcgt gggccccgag tgcttcggcc tgatctctaa ccagtttagc atcatcggcc 1201 acgtgatcgg cgacgtgatc agcaagaatc tgtggggcct gatggcctgg aagctgcgcc 1261 tgaagatcaa ggatggccag cccatggagc acgtgaaccc tctgaagatc gtgagcaacg 1321 atggcactaa tactatggaa aactatcccc tggacaaaat cattcagacc ggaccggtgg 1381 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 1441 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 1501 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 1561 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 1621 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 1681 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg 1741 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 1801 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 1861 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 1921 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 1981 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 2041 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagag ctcaagcttc 2101 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat 2161 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 2221 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 2281 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 2341 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg 2401 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 2461 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 2521 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 2581 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 2641 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 2701 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 2761 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 2821 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 2881 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 2941 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg 3001 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3061 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 3181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 3241 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 3301 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 3361 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3421 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3481 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3541 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3601 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3661 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3721 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3781 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3841 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 3901 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3961 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4021 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4081 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4141 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4201 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4261 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4321 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 4381 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 4441 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 4501 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 4561 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 4621 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 4681 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 4741 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 4801 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 4861 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 4921 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 4981 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 5041 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 5101 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 5161 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 5221 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 5281 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 5341 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 5401 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 5461 taccgccatg cattagttat taatagtaa //