LOCUS Exported 5621 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-MerMAID7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5621) 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 5621) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5621 /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 1512..2219 /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(1654..1672) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1922..1941 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2393..2514 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2430..2449) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2484..2503 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2521..2976) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2658..2679) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2870..2889 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3003..3107 /gene="bla" /label=AmpR promoter promoter 3109..3466 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3132..3152) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3317..3452 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3379..3398 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3501..4295 /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(3555..3574) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4165..4184 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4483..4502) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4527..4574 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4903..5491 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5392..5411 /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 tccgtggaac 601 tgcatgtctt tgcccagatt ttcactgcct tcgtgtcagc cgccaacgcc ctgtattact 661 tctatatcta caggtgcacc aagagaggcc acctgatgaa gtgggaggtg gcatgggtgg 721 ccagcgtgga gacagtgaac tacgtgatcc aggtggccac aggctccccc cgggtggagc 781 tggcccacgg aggcatgttc ccttggatgc gctatgtggg ctggcagctg acctgtccag 841 tgctgctggc ctttatcgtg gtgaatatgc tggagagccc ctccacaagg ctggtcatcc 901 agatgctgat gctgctgcag ttcatcatcc tgtctggcat gaccgccagc ctggtggccg 961 atatgtccat gaagatcctg tttgtggtca tcgccgccct gtctctgtgc atcctgtact 1021 atcagctgta tacccaccgg aagtataagg acgtgcctac cacacacaac aaggagtacc 1081 acctgctgct gtatttcatg ggctcttggc tggtgttccc catcctgttt atcgtgggcc 1141 ctgagatgac caatctggtg gccttcgagc tgacactggt gggccactgc gtgggcgatc 1201 tgatcgccaa gaacgccttt ggcctgctgg cctggcagta caccaagtac gtgtataaga 1261 agaatctgaa ggcacctgac ctggaggcac acgcagcacc agatctggag gcccacagcg 1321 cccaggacct ggagcaggtg gcagtggcaa acccaagggc caagacagac ctgccccaga 1381 ccatcacact gcccctgcac atgtttcctt atatgctgtc tccaaagagc gcccactccc 1441 ccagatacac tgatacaact actccaagcg aaaaaagctc tggggaaaag tcttctgggg 1501 aaggaccggt ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc 1561 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg 1621 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc 1681 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg 1741 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt 1801 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc 1861 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg 1921 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg 1981 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact 2041 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct 2101 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac 2161 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag 2221 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag 2281 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct 2341 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt 2401 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 2461 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt 2521 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 2581 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 2641 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 2701 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 2761 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 2821 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 2881 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 2941 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg 3001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 3061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa 3121 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3301 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3361 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 3421 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga 3481 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3541 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3601 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3661 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg 3721 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 3781 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 3841 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 3901 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 3961 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4021 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4081 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4141 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4201 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4261 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 4321 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4381 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4441 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac 4501 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt 4561 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc 4621 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc 4681 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc 4741 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 4801 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 4861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 4921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 4981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 5041 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 5101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 5161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 5221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 5281 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 5341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 5401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 5461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 5521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 5581 ggataaccgt attaccgcca tgcattagtt attaatagta a //