LOCUS Exported 5758 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Anion-conducting channelrhodopsin of viral origin. Codon-optimized for mammalian expression (human/mouse).. ACCESSION . VERSION . KEYWORDS vPyACR_21821 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5758) AUTHORS Rozenberg A, Oppermann J, Wietek J, Lahore RGF, Sandaa RA, Bratbak G, Hegemann P, Beja O TITLE Lateral Gene Transfer of Anion-Conducting Channelrhodopsins between Green Algae and Giant Viruses JOURNAL Current Biology. 2020. 30: 1–11 REFERENCE 2 (bases 1 to 5758) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5758 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 80..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 384..587 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 538..558 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 628..684 /label=MCS /note="multiple cloning site" regulatory 685..694 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1717..2430 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1759..1780) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2020..2039) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2367..2388 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2565..2686 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2602..2621) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2656..2675 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2693..3148) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2830..2851) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3042..3061 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3175..3279 /gene="bla" /label=AmpR promoter promoter 3281..3638 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3304..3324) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3489..3624 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3551..3570 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3673..4467 /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(3727..3746) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4337..4356 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4655..4674) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4699..4746 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5075..5663 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5564..5583 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc 601 gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttctgcagtc 661 gacggtaccg cgggcccggg atccgccacc atggactatg ggggagcact gtccgccgtc 721 gggctgtttc agacttccta caccctggag aacaacggga gcgtcatttg cattcctaac 781 aatggccagt gcttctgtct ggcctggctg aagagcaacg ggatgtcctt tgaccagacc 841 atttgtaagc atattcagtt tattgacctg ctgctgtcca ttatcctgtt catcttcttc 901 tattatcaga agatcgtgaa gaggagatgc aagttcgagg tgctgtacgt gaccggcgtg 961 ggctctctga cagacctggt gaagctgacc atcacaaatg agctgccagc ctcctatgtg 1021 atcgagaaca atggcgtgct gttcaactgg tctcagtttc tgggctggat gatcacctgc 1081 cccgtgctgc tgatccacct gagcaatctg gccggcaagg acgtgttcga tgtgcggcgc 1141 atgatgaaga tcctggtcat ctaccagatc ctgatggtgt ccggcgccac agcctctatg 1201 tgcaacaatg atctgaagtg gctgttcttt ggcgtgtctc tgatcagcct gctgaagatc 1261 tacatctatg cccacaacat ctttcaggag agccgggaga tcatgcctaa gcgcgccaac 1321 gtggtcatca actggatctc cttcgtgttt tacgtgagct ggtccggctt cggcgtgagc 1381 tggttctttt ccccaaatgg catctccttc ttttctgact acaccacaaa ggccctgttt 1441 gccttctttg atatcatgag caagaacgtg tattccatgc tgggctggta tctgaggtgg 1501 tatatcatca gaaagttcga caatcccgag gagtttgtgg agcagtacca ggaggaggat 1561 gagaagaaga tcaacatcct gctggtggag aacaatgcca tctactccca ctatttcgac 1621 aatatcctga tccagcacgg ctgtaccgtg gacgtgaaga gcaggatcac cagcgagggc 1681 gagtacatcc ccctggacca gatcgacatc aacgtggtga gcaagggcga ggagctgttc 1741 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 1801 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 1861 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg 1921 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 1981 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 2041 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 2101 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 2161 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 2221 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 2281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc 2341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 2401 atcactctcg gcatggacga gctgtacaag ttctgctacg agaacgaggt gtgatctaga 2461 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 2521 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 2581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 2641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta 2701 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac 2761 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 2821 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 2881 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 2941 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 3001 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 3061 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 3121 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga 3181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 3241 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc 3301 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3361 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3541 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 3601 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga 3661 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 3721 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 3781 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 3841 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 3901 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 3961 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 4021 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 4081 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 4141 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 4201 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 4261 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 4321 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 4381 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 4441 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 4501 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 4561 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 4621 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg 4681 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt 4741 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 4801 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 4861 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt 4921 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 4981 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 5041 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 5101 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 5161 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 5221 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 5281 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 5341 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 5401 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 5461 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 5521 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 5581 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 5641 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 5701 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataa //